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P16284 , P05164 and P10644 at chromosome 17q21-q24 and susceptibility for multiple sclerosis in Sweden and Sardinia . Using genome screen , DNA sequence and mapping data , we scanned the human chromosomal region 17q21-q24 for polymorphic markers in single copy genes . Three such genes were identified : the gene for myeloperoxidase ( P05164 ) at 17q21.3-q23.2 , containing a CA-microsatellite in the eighth intron and a functional single base substitution ( G to A ) in the promoter region , the platelet endothelial cell adhesion molecule-1 gene ( P16284 ) at 17q23 , which has a CA-repeat sequence in the sixth intron , and the gene for the regulatory subunit RIalpha of DB02527 -dependent protein kinase ( P10644 ) at 17q23-q24 , in which a GA-microsatellite was detected in the 5'-flanking region . Association of these polymorphisms with multiple sclerosis ( MS ) was studied in a Swedish case-control population of 199 MS patients and 145 control subjects , and in 203 simplex families from Sardinia . None of these polymorphic genes was found to be a genetic marker for disease susceptibility . These results are in contrast with previous studies on the involvement of P05164 in MS and suggest that the elevated expression of P16284 in MS , as earlier documented , is related to transactivation by other gene products . The effect of anti- P15692 drugs ( bevacizumab and aflibercept ) on the survival of patients with metastatic colorectal cancer ( mCRC ) . Significant progression has been achieved in the treatment of metastatic colorectal cancer ( mCRC ) in recent years . This has been partly attributed to successfully incorporating new drugs into combination chemotherapy . In addition to the traditional cytotoxic chemotherapeutic agents , molecularly targeted agents began to play an important role in the treatment of advanced solid tumors . To date , two classes of molecularly targeted agents have been approved for treatment of patients with mCRC : ( 1 ) antivascular endothelial growth factor ( anti- P15692 ) agents ( such as bevacizumab and aflibercept ) and ( 2 ) antiendothelial cell growth factor receptor ( anti- P00533 ) agents ( such as cetuximab and panitumumab ) . DB08885 is a new member of anti- P15692 agents which has demonstrated efficacy for treatment of mCRC . With the commencement of clinical trials and basic research into aflibercept , more data from the bedside and the bench have been obtained . This review will outline the application of anti- P15692 agents by reviewing clinic experiences of bevacizumab and aflibercept , and try to add perspectives on the use of anti- P15692 agents in mCRC . Randomised phase II study of siltuximab ( CNTO 328 ) , an anti- P05231 monoclonal antibody , in combination with mitoxantrone/prednisone versus mitoxantrone/prednisone alone in metastatic castration-resistant prostate cancer . PURPOSE : This open-label phase II trial assessed mitoxantrone/prednisone ( M/P ) with and without siltuximab ( CNTO 328 ) , an anti-interleukin-6 chimeric monoclonal antibody , for patients with metastatic castration-resistant prostate cancer who received prior docetaxel-based chemotherapy . METHODS : Part 1 assessed the safety of biweekly siltuximab 6 mg/kg plus M 12 mg/m(2) every 3 weeks and P. Part 2 assessed efficacy and safety of siltuximab plus M/P versus M/P alone . The primary end-point was progression-free survival ( PFS ) . Progression was defined as progressive disease per Response Evaluation Criteria in Solid Tumours ( RECIST ) , or ≥ 3 new skeletal lesions with clinical deterioration or without deterioration confirmed by repeated bone scan . Rising prostate-specific antigen was not considered progression . RESULTS : DB09036 plus M/P was well tolerated in Part 1 ( n=9 ) . In Part 2 , 48 and 49 patients received siltuximab plus M/P or M/P alone , respectively . Enrolment was prematurely terminated by the Independent Data Monitoring Committee since an apparent imbalance in patient baseline characteristics ( favoring the M/P only arm ) made it unlikely that the study could achieve its primary efficacy end-point . Median PFS was 97 days with siltuximab combination and 228 days with M/P alone ( hazard ratio , 1.72 ; P=0.043 ) . Use of a novel non-validated PFS definition may have contributed to this result . Abnormal laboratory assessments were more frequent with the combination . Infection and febrile neutropenia rates were similar between groups . Greater P02741 suppression was achieved during siltuximab combination treatment compared with M/P alone ( P=0.0003 ) . CONCLUSION : While siltuximab plus M/P appeared well tolerated , improvement in outcomes was not demonstrated . Consequences of missense mutations for dimerization and turnover of alanine:glyoxylate aminotransferase : study of a spectrum of mutations . DB00160 :glyoxylate aminotransferase ( AGT ) is a liver peroxisomal enzyme , deficiency of which results in primary hyperoxaluria type 1 ( P78364 ) . More than 65 P78364 -related mutations are now documented in the AGT gene ( P21549 ) , of which about 50 % are missense . We have generated a spectrum of 15 missense changes including the most common P78364 mutation , G170R , and expressed them on the appropriate background of the major or minor allele , in an Escherichia coli overexpression system and in a rabbit reticulocyte transcription/translation system . We have investigated their effects on enzyme activity , dimerization , aggregation , and turnover . The effect of pyridoxal phosphate ( PLP ) on dimerization and stability was also investigated . Although all 15 mutant AGTs were expressed as intact proteins in E. coli , only three : G41R and G41V on the major allele , and the common mutation G170R , resulted in significant amounts of enzymatic activity . Dimerization failure was a frequent observation ( 13/15 ) except for G41V and D183N . Dimerization was poor with S187F but was substantially improved with PLP . Proteasome-mediated protein degradation was observed for all the mutations except G41R on the major allele , G41V , D183N , G170R , and S218L . Increases in the stability of the mutant enzymes in the presence of PLP were small ; however , G41R on the minor allele showed a direct relationship between its half life and the concentration of PLP . The minor allele AGT product and many of the mutants were subject to a limited non-proteasomal proteolytic cleavage when DB00171 was depleted . The roles of cyclic nucleotide phosphodiesterases ( PDEs ) in steroidogenesis . The second messenger , DB02527 , is one of the most important regulatory signals for control of steroidogenesis . This review focuses on current knowledge about regulation of cyclic nucleotides by phosphodiesterases ( PDEs ) in steroidogenic tissues . The first PDE known to directly regulate steroidogenesis was PDE2 , the cGMP-stimulated PDE . PDE2 mediates P01160 /cGMP-induced decreases in aldosterone production . Recently , the PDE8 family has been shown to control steroidogenesis in two tissues . Specifically , O60658 regulates testosterone production by itself and in concert with additional DB07954 -sensitive PDEs . O95263 modulates basal corticosterone synthesis via acute and chronic mechanisms . In addition to DB02527 -dependent pathways , cGMP signaling also can promote steroidogenesis , and O76074 modulates this process . Finally , PDE mutations may lead to several human diseases characterized by abnormal steroid levels . Effects of MLN518 , a dual P36888 and P10721 inhibitor , on normal and malignant hematopoiesis . Internal tandem duplications ( ITDs ) of the P07333 -like tyrosine kinase 3 ( P36888 ) receptor tyrosine kinase are found in approximately 30 % of patients with acute myelogenous leukemia ( AML ) and are associated with a poor prognosis . P36888 ITD mutations result in constitutive kinase activation and are thought to be pathogenetically relevant , implicating P36888 as a plausible therapeutic target . MLN518 ( formerly CT53518 ) is a small molecule inhibitor of the P36888 , P10721 , and platelet-derived growth-factor receptor ( P09619 ) tyrosine kinases with significant activity in murine models of P36888 ITD-positive leukemia . Given the importance of P36888 and P10721 for normal hematopoietic progenitor cells , we analyzed the effect of MLN518 on murine hematopoiesis under steady-state conditions , after chemotherapy-induced myelosuppression , and during bone marrow transplantation . In these assays , we show that MLN518 has mild toxicity toward normal hematopoiesis at concentrations that are effective in treating P36888 ITD-positive leukemia in mice . We also demonstrate that MLN518 preferentially inhibits the growth of blast colonies from P36888 ITD-positive compared with ITD-negative patients with AML , at concentrations that do not significantly affect colony formation by normal human progenitor cells . In analogy to imatinib mesylate in P11274 - P00519 -positive acute leukemia , MLN518-induced remissions may not be durable . Our studies provide the basis for integrating this compound into chemotherapy and transplantation protocols . The expanding role of vascular endothelial growth factor inhibitors in ophthalmology . Vascular endothelial growth factor ( P15692 ) plays an important role in both physiologic and pathologic angiogenesis and contributes to increased permeability across both the blood-retinal and blood-brain barriers . After 2 decades of extensive research into the P15692 families and receptors , specific molecules have been targeted for drug development , and several medications have received US Food and Drug Administration approval . DB00112 , a full-length antibody against P15692 approved for the intravenous treatment of advanced carcinomas , has been used extensively in ophthalmology for exudative age-related macular degeneration , diabetic retinopathy , retinal vein occlusions , retinopathy of prematurity , and other chorioretinal vascular disorders . DB04895 and ranibizumab have been developed specifically for intraocular use , whereas the soon-to-be-introduced aflibercept ( DB08885 ) is moving through clinical trials for both intraocular and systemic use . Although these drugs exhibit excellent safety profiles , ocular and systemic complications , particularly thromboembolic events , remain a concern in patients receiving therapy . Patients experiencing adverse events that may be related to P15692 suppression should be carefully evaluated by both the ophthalmologist and the medical physician to reassess the need for intraocular therapy and explore the feasibility of changing medications . For this review a search of PubMed from January 1 , 1985 through April 15 , 2011 , was performed using the following terms ( or combination of terms ) : vascular endothelial growth factors , P15692 , age-related macular degeneration , diabetic retinopathy , retina vein occlusions , retinopathy of prematurity , intravitreal injections , bevacizumab , ranibizumab , and DB08885 . Studies were limited to those published in English . Other articles were identified from bibliographies of retrieved articles and archives of the author . Phase 1 trial of linifanib ( DB06080 ) in patients with refractory or relapsed acute myeloid leukemia . Linifanib , a potent oral inhibitor of fms-like tyrosine kinase 3 ( P36888 ) and vascular endothelial growth factor receptor tyrosine kinases , has demonstrated promising preclinical single-agent and synergistic anti-leukemic activity in combination with cytarabine . In this phase 1 , multicenter , open-label , dose-escalation study , 45 adults with relapsed/refractory acute myeloid leukemia ( AML ) received linifanib alone in arm A ( n = 29 ) and linifanib plus intermediate-dose cytarabine in arm B ( n = 16 ) . Median treatment duration was 21 days ( range 5-110 ) . Linifanib was well tolerated overall . The most common grade 3/4 events were fatigue ( arm A ) and febrile neutropenia ( arm B ) . The recommended phase 2 dose was 15 mg ( alone ) , and 10 mg ( with cytarabine ) . Evidence of on-target kinase inhibition in patients with P36888 -mutant and wild-type AML was seen . Decreased phosphorylated P36888 was seen in 3/3 patients with P36888 -internal tandem duplication ( ITD ) with peripheral blast reductions and in 8/24 ( 33 % ) patients with wild-type , D835 or unknown P36888 mutation . Eight/29 ( 28 % ) patients had decreased phosphorylated extracellular signal-regulated kinase ( P29323 ) . Establishment of a double Philadelphia chromosome-positive acute lymphoblastic leukemia-derived cell line , TMD5 : effects of cytokines and differentiation inducers on growth of the cells . A double Philadelphia chromosome ( Ph ) -positive leukemia cell line with common-B cell phenotype , designated TMD5 , was established from the blast cells of a patient with double Ph-positive acute lymphoblastic leukemia . TMD5 cells expressed 190 kDa P11274 / P00519 chimeric protein and 145 kDa P00519 protein . The cells proliferated without added growth factors . Autocrine growth mechanism was not recognized . The addition of growth factors such as DB00099 , GM- P04141 , P08700 , P05231 , or Stem Cell Factor did not affect the growth . Herbimycin A suppressed the growth of TMD5 cells at the low concentration that did not affect Ph-negative cells . It suppressed tyrosine phosphorylation of intracellular proteins in TMD5 cells . Dexamethasone and dibutyryl cyclic AMP also suppressed the growth . They , however , did not affect the phosphorylation significantly . Neither all-trans retinoic acid nor interferon-alpha affected the growth . TMD5 cells , characterized minutely here and rare in that they have double Ph chromosomes , will be a useful tool for the study of Ph-positive leukemia . Interaction between fatty acid synthase- and ErbB-systems in ovarian cancer cells . P49327 ( P49327 ) represents a metabolic oncogene . It produces phospholipids for membrane microdomains that accommodate receptor tyrosine kinases including Epidermal Growth Factor-Receptor ( P00533 , ErbB1 ) and ErbB2 ( P04626 /neu ) . P49327 and ErbBs are overexpressed in ovarian cancer . We examined the effect of P49327 and ErbB inhibition on A2780 and SKOV3 ovarian cancer cells . Growth assays reveal that P49327 inhibitor C75 sensitizes tumor cells against anti-ErbB drugs ( pelitinib [ Q9Y259 -569 ] , canertinib [ DB05424 ] , erlotinib , cetuximab , matuzumab , trastuzumab ) suggesting P49327 /ErbB cooperation . qRT-PCR and Western blotting revealed that C75 represses P49327 , P00533 , ErbB2 , and AKT suggesting that P49327 -induced membrane microdomains accommodate/stabilize ErbBs and facilitate AKT recruitment/activation . Our data indicate that AKT is crucial for ErbB/ P49327 interaction , AKT cross-inhibits P29323 and feeds loops that boost P49327 and P00533 transcription , and P00533 and ErbB2 must be co-silenced for maximal P49327 downregulation . Taken together , interference with P49327 and ErbB abrogates their oncogenicity and should be exploited for ovarian cancer treatment . DB09559 , a fully human IgG1 mAb directed against the P00533 for the potential treatment of cancer . DB09559 ( DB05774 ) , under development by ImClone Systems in collaboration with Bristol-Myers Squibb , is a fully human IgG1 mAb targeting the epidermal growth factor receptor ( P00533 ) , for the potential intravenous treatment of cancer , in particular NSCLC . In vitro studies demonstrate that necitumumab inhibits downstream targets in the P00533 pathway ( eg , MAPK ) , which are important for cellular proliferation , differentiation , invasion and metastasis . Furthermore , because necitumumab is an IgG1 construct , it has the potential to induce antibody-dependent cell-mediated cytotoxicity against tumor cells . Preclinical studies indicated that the antitumor activity of necitumumab is either comparable with or superior to that of ImClone 's chimeric anti- P00533 mAb cetuximab . In a phase I clinical trial in patients with advanced solid malignancies , necitumumab displayed nonlinear pharmacokinetic behavior . The toxicity profile of necitumumab is acceptable , with skin toxicity being the most frequently reported adverse event in the phase I and II clinical trials conducted to date . Preliminary data from a phase II clinical trial of necitumumab in combination with chemotherapy for the first-line treatment of advanced colon cancer are promising . Success in the ongoing phase III clinical trials in patients with advanced NSCLC would lead to necitumumab becoming a valuable addition to future therapeutic strategies in oncology . Preclinical rationale for combining an P00533 antibody with cisplatin/gemcitabine for the treatment of NSCLC . BACKGROUND : Although the addition of epidermal growth factor receptor ( P00533 ) antibodies to various platinum-based chemotherapy regimens for non-small cell lung cancer ( NSCLC ) is being actively pursued in the clinic , rationale for the prioritization of specific regimens is lacking . MATERIALS AND METHODS : We evaluated the antitumor effects of necitumumab , a recombinant human IgG1 antibody targeting P00533 , in combination with cisplatin plus gemcitabine , pemetrexed , or paclitaxel in a panel of 9 subcutaneous tumor models of NSCLC established in nu/nu athymic mice . RESULTS : DB09559 in combination with cisplatin/gemcitabine was particularly effective , although interestingly , the mechanisms underlying these benefits were model dependent . For example , increased tumor cell apoptosis contributed towards combination efficacy in the A549 model , in association with increased expression of hsa-miR-29b and reduced expression of antiapoptotic genes including DNA methyltransferase Q9UBC3 , commonly up-regulated in patients with NSCLC . Such inverse effects of combination therapy on Q9UBC3 and hsa-miR-29b expression were found in multiple models . Importantly , in the A549 model , hsa-miR-29b down-regulation of DMNT3b reduced promoter methylation of tumor suppressor genes such as Q9BY67 ( Q9BY67 ) , Ras associated ( Q12967 /AF-6 ) domain family member 1 ( Q9NS23 ) , and Fragile histidine triad gene ( P49789 ) , increasing their expression . CONCLUSION : These results offer a preclinical rationale for combining an P00533 antibody with cisplatin/gemcitabine for patients with NSCLC , and provide potential molecular biomarkers for tailoring therapy . Individual cerebellar Purkinje cells express different cGMP phosphodiesterases ( PDEs ) : in vivo phosphorylation of cGMP-specific PDE ( O76074 ) as an indicator of cGMP-dependent protein kinase ( PKG ) activation . The nitric oxide ( NO ) -cGMP pathway has been implicated as playing a crucial role in the induction of cerebellar long-term depression ( LTD ) . The amplitude and duration of the cGMP signal is controlled by cyclic nucleotide phosphodiesterases ( PDEs ) . Here we identify O76074 and Q01064 as the two major cGMP-hydrolyzing PDEs specifically and differentially expressed in the Purkinje neurons of mouse cerebellum . O76074 was found in all Purkinje neurons , whereas Q01064 was detected only in a subset of these cells , suggesting that individual Purkinje cells may differentially regulate cGMP , depending on the PDE isozymes expressed . Although expression of guanylate cyclase and/or cGMP-dependent protein kinase ( PKG ) in Purkinje cells have been reported , neither cGMP accumulation nor PKG activation in these cells in vivo has been demonstrated . To determine if changes in PKG activation and O76074 regulation occur in vivo we have examined the phosphorylation of O76074 in mouse cerebellar Purkinje cells by immunocytochemistry and Western blot analyses using a phosphospecific O76074 antibody . Injection of sodium nitroprusside or selective PKG activators into the lateral ventricle of mouse brain induced O76074 phosphorylation in vivo , but was completely missing in Purkinje cell-specific PKG I knock-out mice . In cerebellar slices , treatment with sildenafil or DB07954 led to different levels of phospho- O76074 accumulation and activation of O76074 . These results suggest that phosphorylation of O76074 in Purkinje neurons after cGMP-PKG activation performs a critical role in the termination of the cGMP signal during LTD progression ; moreover , O76074 phosphorylation may be used as an in vivo indicator for PKG activation . P12931 -family kinases are activated in non-small cell lung cancer and promote the survival of epidermal growth factor receptor-dependent cell lines . The role of Src-family kinases ( SFKs ) in non-small cell lung cancer ( NSCLC ) has not been fully defined . Here we addressed this question by examining SFK phosphorylation in NSCLC biopsy samples and using genetic and pharmacological approaches to inhibit SFK expression and activity in cultured NSCLC cells . Immunohistochemical analysis of NSCLC biopsy samples using a Tyr416 phosphorylation-specific , pan-SFK antibody revealed staining in 123 ( 33 % ) of 370 tumors . Because c-Src is known to be both an upstream activator and downstream mediator of epidermal growth factor receptor ( P00533 ) , we next investigated SFK phosphorylation in a panel of NSCLC cell lines , including ones that depend on P00533 for survival . The P00533 -dependent NSCLC cell lines HCC827 and H3255 had increased phosphorylation of SFKs , and treatment of these cells with an SFK inhibitor ( P50391 or DB06616 ) induced apoptosis . P50391 decreased phosphorylation of P00533 , ErbB2 , and ErbB3 and strikingly enhanced apoptosis by gefitinib , an P00533 inhibitor . HCC827 cells transfected with c-Src short hairpin RNA exhibited diminished phosphorylation of P00533 and ErbB2 and decreased sensitivity to apoptosis by P50391 or gefitinib . We conclude that SFKs are activated in NSCLC biopsy samples , promote the survival of P00533 -dependent NSCLC cells , and should be investigated as therapeutic targets in NSCLC patients . P04626 kinase domain mutation results in constitutive phosphorylation and activation of P04626 and P00533 and resistance to P00533 tyrosine kinase inhibitors . P04626 /Neu gene mutations have been identified in lung cancer . Expression of a P04626 mutant containing a G776(YVMA) insertion in exon 20 was more potent than wild-type P04626 in associating with and activating signal transducers , phosphorylating P00533 , and inducing survival , invasiveness , and tumorigenicity . P04626 (YVMA) transphosphorylated kinase-dead P00533 (K721R) and P00533 (WT) in the presence of P00533 tyrosine kinase inhibitors ( TKIs ) . Knockdown of mutant P04626 in H1781 lung cancer cells increased apoptosis and restored sensitivity to P00533 TKIs . The P04626 inhibitors lapatinib , trastuzumab , and DB05424 inhibited growth of H1781 cells and cells expressing exogenous P04626 (YVMA) . These data suggest that ( 1 ) P04626 (YVMA) activates cellular substrates more potently than P04626 (WT) ; and ( 2 ) cancer cells expressing this mutation remain sensitive to P04626 -targeted therapies but insensitive to P00533 TKIs . Disruption of Src function potentiates Chk1-inhibitor-induced apoptosis in human multiple myeloma cells in vitro and in vivo . Ras/MEK/ P29323 pathway activation represents an important compensatory response of human multiple myeloma ( MM ) cells to checkpoint kinase 1 ( Chk1 ) inhibitors . To investigate the functional roles of Src in this event and potential therapeutic significance , interactions between Src and Chk1 inhibitors ( eg , P55089 -01 or Chk1i ) were examined in vitro and in vivo . The dual Src/Abl inhibitors BMS354825 and DB06616 blocked Chk1-inhibitor-induced extracellular signal-regulated kinase 1/2 ( P27361 /2 ) activation , markedly increasing apoptosis in association with BimEL up-regulation , p34(cdc2) activation , and DNA damage in MM cell lines and primary CD138(+) MM samples . Loss-of-function Src mutants ( K297R , K296R/Y528F ) or shRNA knock-down of Src prevented the P27361 /2 activation induced by Chk1 inhibitors and increased apoptosis . Conversely , constitutively active Ras or mitogen-activated protein kinase/ P29323 kinase 1 ( Q02750 ) significantly diminished the ability of Src inhibitors to potentiate Chk1-inhibitor lethality . Moreover , Src/Chk1-inhibitor cotreatment attenuated MM-cell production of vascular endothelial growth factor and other angiogenic factors ( eg , P03950 [ angiogenin ] , P01033 /2 [ tissue inhibitor of metalloproteinases 1/2 ] , and RANTES [ regulated on activation normal T-cell expressed and secreted ] ) , and inhibited in vitro angiogenesis . Finally , coadministration of BMS354825 and P55089 -01 suppressed human MM tumor growth in a murine xenograft model , increased apoptosis , and diminished angiogenesis . These findings suggest that Src kinase is required for Chk1-inhibitor-mediated Ras → P27361 /2 signaling activation , and that disruption of this event sharply potentiates the anti-MM activity of Chk1 inhi-bitors in vitro and in vivo . Protein kinase A alterations in endocrine tumors . Various molecular and cellular alterations of the cyclic adenosine monophosphate ( DB02527 ) pathway have been observed in endocrine tumors . Since protein kinase A ( PKA ) is a central key component of the DB02527 pathway , studies of the alterations of PKA subunits in endocrine tumors reveal new aspects of the mechanisms of DB02527 pathway alterations in human diseases . So far , most alterations have been observed for the regulatory subunits , mainly P10644 and to a lower extent , P31323 . One of the best examples of such alteration today is the multiple neoplasia syndrome Carney complex ( CNC ) . The most common endocrine gland manifestations of CNC are pituitary GH-secreting adenomas , thyroid tumors , testicular tumors , and DB01285 -independent Cushing 's syndrome due to primary pigmented nodular adrenocortical disease ( PPNAD ) . Heterozygous germline inactivating mutations of the PKA regulatory subunit RIα gene ( P10644 ) are observed in about two-third of CNC patients , and also in patients with isolated PPNAD . P10644 is considered as a tumor suppressor gene . Interestingly , these mutations can also be observed as somatic alterations in sporadic endocrine tumors . More than 120 different P10644 mutations have been found today . Most of them lead to an unstable mutant mRNA , which will be degraded by nonsense mediated mRNA decay . In vitro and in vivo functional studies are in progress to understand the mechanisms of endocrine tumor development due to PKA regulatory subunits inactivation . P10644 mutations stimulate in most models PKA activity , mimicking in some way DB02527 pathway constitutive activation . Cross-talks with other signaling pathways summarized in this review have been described and might participate in endocrine tumorigenesis . Differential sensitivity to cardiotonic drugs of cyclic AMP phosphodiesterases isolated from canine ventricular and sinoatrial-enriched tissues . A cardiac phosphodiesterase ( PDE ) which specifically hydrolyzes DB02527 and is inhibited by cyclic GMP has been suggested to be the site of action of new cardiotonic drugs . To investigate the effect of inhibitors , canine cyclic nucleotide PDEs were isolated from left ventricle and from sinoatrial node-enriched tissue , using identical techniques . Four PDE forms could be chromatographically resolved from each tissue , including a peak I PDE ( calmodulin-activated phosphodiesterase , P62158 -PDE ) , a peak II PDE ( cyclic GMP-stimulated phosphodiesterase , O00408 ) and a peak III PDE ( specific for cyclic AMP ) . The latter was further fractionated into two forms : One was inhibited by cyclic GMP and by the platelet antiaggregant AAL 05 ( CGI-PDE ) , and the second was insensitive to cyclic GMP and was inhibited by rolipram ( ROI-PDE ) . Reference PDE inhibitors , isobutyl-1-methylxanthine ( DB07954 ) and papaverine , nonselectively inhibited the four forms isolated from the two tissues . Cardiotonic drugs ( CI 930 , LY 181512 , piroximone , enoximone , and SK & F 94120 ) selectively inhibited CGI-PDE from ventricular tissue but were poorly active on both CGI-PDE and ROI-PDE from the sinoatrial-enriched fraction . In contrast , milrinone inhibited CGI-PDEs and ROI-PDEs from both ventricular and sinoatrial tissues . These results are in good agreement with pharmacologic data in the literature on the positive chronotropic and inotropic effects of the studied drugs in the dog . They provide a possible basis for the dissociation of these two properties of PDE inhibitors . K-Ras gene status and expression of Ras/mitogen-activated protein kinase ( MAPK ) signaling molecules in ameloblastomas . BACKGROUND : To clarify the roles of rat sarcoma ( Ras ) /mitogen-activated protein kinase ( MAPK ) signaling pathway in oncogenesis and cytodifferentiation of odontogenic tumors , K-Ras gene status and expression of Ras , Raf1 , MAPK/extracellular signal-regulated kinase ( P29323 ) kinase (MEK)1 , and P27361 /2 proteins were analyzed in ameloblastomas as well as in tooth germs . METHODS : Paraffin sections of 10 tooth germs and 46 benign and 6 malignant ameloblastomas were examined immunohistochemically for the expression of K-Ras , Raf1 , Q02750 , and P27361 /2 . Frozen tissue samples of 22 benign ameloblastomas and 1 malignant ( metastasizing ) ameloblastoma were analyzed by direct DNA sequencing to detect K-Ras gene alteration . RESULTS : Immunohistochemical reactivity for K-Ras , Raf1 , Q02750 , and P27361 /2 was detected in both normal and neoplastic odontogenic epithelium , and these molecules were reactive chiefly with odontogenic epithelial cells neighboring the basement membrane . Plexiform ameloblastomas showed slightly stronger expression of these Ras/MAPK signaling molecules than follicular ameloblastomas . Keratinizing cells and granular cells showed decreased reactivity for the signaling molecules . Basal cell ameloblastomas showed slightly stronger reactivity for the signaling molecules than did the other subtypes . K-Ras immunoreactivity in malignant ameloblastomas was lower than that in dental lamina of tooth germs . Direct DNA sequencing showed a P19440 to GCT point mutation at codon 12 of K-Ras gene in one ameloblastoma . CONCLUSION : Expression of K-Ras , Raf1 , Q02750 , and P27361 /2 in tooth germs and ameloblastomas suggests that Ras/MAPK signaling pathway functions to regulate cell proliferation and differentiation in both normal and neoplastic odontogenic epithelium . K-Ras gene status implied that K-Ras mutations might play a minor role in oncogenesis of odontogenic epithelium . Gene transfer of GM- P04141 , P33681 and CD154 cDNA enhances survival in a murine model of acute leukemia with persistence of a minimal residual disease . Gene transfer of various cytokines and co-stimulatory molecules has been reported to induce a potent antileukemic immunity in murine models , however , the relative efficiency and possible synergistic effects between candidate genes have not been extensively investigated . We analyzed in a murine model of P11274 / P00519 acute leukemia whether gene transfer of CD154 , P33681 or GM- P04141 as a single agent or combination of CD154 + GM- P04141 , P33681 + CD154 and GM- P04141 + P33681 in leukemic cells could enhance survival . We observed that CD154 gene transfer induced a marked inhibition of leukemogenicity , and also that CD154 and combination of GM- P04141 and P33681 gene transfer protected mice against subsequent challenge with leukemic cells and had a therapeutic effect for a pre-established leukemia disease . We also found minimal residual leukemic disease by RT-PCR for 6 to 12 months in 0 to 25 % of animals injected with transduced leukemic cells and surviving the challenge without evidence of disease , except in the control empty plasmid group where very few mice survived the challenge but all of those were positive by RT-PCR . These findings suggest that leukemic cell vaccination by gene transfer can induce a tumor dormancy phenomenon compatible with long-term survival . Selective up-regulation of PDE1B2 upon monocyte-to-macrophage differentiation . P04141 ( GM- P04141 ) is a major regulator of monocyte to macrophage differentiation . In both humans and mice , the main phenotype of decreased GM- P04141 function is pulmonary proteinosis due to aberrant function of alveolar macrophages . Recently , this cytokine has been shown to up-regulate a cyclic nucleotide phosphodiesterase , Q01064 . Two Q01064 variants with unique N-terminal sequences , Q01064 and PDE1B2 , have been identified . Here , we report that the previously uncharacterized PDE1B2 is selectively increased by GM- P04141 by stimulation of transcription at a previously unknown transcriptional start site . Analysis of the exon and intron organization of the Q01064 gene reveals that PDE1B2 has a different N-terminal sequence because of a separate first exon that is located 11.5 kb downstream from the Q01064 first exon . By using 5'-RACE , alignment of EST sequences , and a luciferase-reporter system , we provide evidence that PDE1B2 has a separate transcriptional start site from Q01064 that can be activated by monocyte differentiation . Furthermore , P05112 treatment in the presence of GM- P04141 , which shifts the differentiation from a macrophage to a dendritic cell phenotype , suppresses the up-regulation of PDE1B2 . Induction of PDE1B2 is also found in T cells upon activation by PHA . Therefore , PDE1B2 may have a regulatory role in multiple immune cell types . Last , characterization of the catalytic properties of recombinant PDE1B2 shows that it prefers cGMP over DB02527 as a substrate and , thus , is likely to regulate cGMP in macrophages . Also , PDE1B2 has a nearly 3-fold lower EC(50) for activation by calmodulin than Q01064 . Meta-analysis of genome-wide association studies confirms a susceptibility locus for knee osteoarthritis on chromosome 7q22 . OBJECTIVES : Osteoarthritis ( OA ) is the most prevalent form of arthritis and accounts for substantial morbidity and disability , particularly in older people . It is characterised by changes in joint structure , including degeneration of the articular cartilage , and its aetiology is multifactorial with a strong postulated genetic component . METHODS : A meta-analysis was performed of four genome-wide association ( GWA ) studies of 2371 cases of knee OA and 35 909 controls in Caucasian populations . Replication of the top hits was attempted with data from 10 additional replication datasets . RESULTS : With a cumulative sample size of 6709 cases and 44 439 controls , one genome-wide significant locus was identified on chromosome 7q22 for knee OA ( rs4730250 , p=9.2 × 10⁻⁹ ) , thereby confirming its role as a susceptibility locus for OA . CONCLUSION : The associated signal is located within a large ( 500 kb ) linkage disequilibrium block that contains six genes : P31323 ( protein kinase , DB02527 -dependent , regulatory , type II , β ) , HPB1 ( HMG-box transcription factor 1 ) , Q9UP83 ( component of oligomeric golgi complex 5 ) , Q99680 ( G protein-coupled receptor 22 ) , O95620 ( dihydrouridine synthase 4-like ) and Q9UHQ4 ( B cell receptor-associated protein 29 ) . Gene expression analyses of the ( six ) genes in primary cells derived from different joint tissues confirmed expression of all the genes in the joint environment . Establishment and phenotypic characterization of human U937 cells with inducible P210 P11274 / P00519 expression reveals upregulation of P13688 ( CD66a ) . Chronic myeloid leukemia ( CML ) is characterized by the expression of the P210 P11274 / P00519 fusion protein . The molecular mechanisms behind this oncogene-mediated hematological disease are , however , not fully understood . Here , we describe the establishment and phenotypic characterization of U937 cells in which P210 P11274 / P00519 can be conditionally expressed using tetracycline . The induction of P11274 / P00519 in the obtained clones resulted in a rapid phosphorylation of the P42224 , P40763 and P42229 molecules , consistent with the findings in other model systems . Phenotypic characterization of the clones revealed that P11274 / P00519 induces a slight decrease in the proliferation and viability , without a marked effect on cell cycle distribution , the rate of apoptosis or on cellular differentiation , as judged by several cell surface markers and capacity to reduce nitro blue tetrazolium . Interestingly , P11274 / P00519 was found to upregulate the expression of carcinoembryonic-related antigen ( P06731 ) P62158 ( CD66a ) , which is a plasma membrane-linked glycoprotein belonging to the CEAs and involved in signal transduction and cellular adhesion . The expression of P13688 was reversible upon imatinib treatment in P11274 / P00519 -expressing U937 cells as well as in P11274 / P00519 -positive K562 cells . The established cell lines may prove useful in further modeling and dissection of P11274 / P00519 -induced leukemogenesis . P23458 activates P40763 activity in non-small-cell lung cancer cells and P05231 neutralizing antibodies can suppress P23458 - P40763 signaling . Members of the signal transducer and activator of transcription ( P35610 ) family of transcription factors are potential targets for the treatment and prevention of cancers including non-small-cell lung cancer . P35610 proteins can be phosphorylated and activated by diverse upstream kinases including cytokine receptors and tyrosine kinases . We examined P35610 protein activation in lung cancer cell lines including those with activating mutations in the P00533 and examined upstream kinases responsible for P40763 phosphorylation and activation using small molecules , antibodies , and RNA interference . We found more pronounced P40763 activation in cells with activating P00533 mutations , yet inhibition of P00533 activity had no effect on P40763 activation . Inhibition of P23458 with small molecules or RNA interference resulted in loss of P40763 tyrosine phosphorylation and inhibition of cell growth . An interleukin-6 neutralizing antibody , siltuximab ( CNTO 328 ) could inhibit P40763 tyrosine phosphorylation in a cell-dependent manner . DB09036 could completely inhibit P40763 tyrosine phosphorylation in H1650 cells , and this resulted in inhibition of lung cancer cell growth in vivo . Combined P00533 inhibition with erlotinib and siltuximab resulted in dual inhibition of both tyrosine and serine P40763 phosphorylation , more pronounced inhibition of P40763 transcriptional activity , and translated into combined effects on lung cancer growth in a mouse model . Our results suggest that P23458 is responsible for P40763 activation in lung cancer cells and that indirect attacks on P23458 - P40763 using an P05231 neutralizing antibody with or without P00533 inhibition can inhibit lung cancer growth in lung cancer subsets . AAV2-mediated gene transfer of P15692 - Q8NHU6 with potent suppression of primary breast tumor growth and spontaneous pulmonary metastases by long-term expression . Vascular endothelial growth factor ( P15692 ) is an important signaling protein and a predominant mediator of angiogenesis in tumor growth and metastasis . Therefore , antagonism of the P15692 pathway results in inhibition of abnormal angiogenesis , then suppression of tumor growth and metastasis . P15692 - Q8NHU6 , a high-affinity soluble decoy receptor , is currently in phase II clinical trails , and has demonstrated more efficacy in different types of solid tumors by intravenous injection every two weeks . In our study , we used recombinant AAV2 as a delivery vehicle to achieve long-lasting expression of DB08885 protein in a mouse model for the first time . We report that AAV2- P15692 - Q8NHU6 can be safely administered and sustained expression in vivo via a single intravenously administration , simultaneously suppressing primary tumor growth and preventing the pulmonary metastases of 4T1 tumors . Decreased microvessel density and increased tumor cell apoptosis were observed in the treatment group . AAV2- P15692 - Q8NHU6 can obviously decrease not only the concentration of P15692 in sera , but also the concentration of other angiogenic factors , such as P05230 , P09038 , angiopoietin-1 and others . These studies suggest that AAV-mediated long-term expression of P15692 - Q8NHU6 is a useful and safe tool to block tumor progression and inhibit spontaneous pulmonary metastases . Role of key residues at the flavin mononucleotide ( Q68DA7 ):adenylyltransferase catalytic site of the bifunctional riboflavin kinase/flavin adenine dinucleotide ( DB03147 ) Synthetase from Corynebacterium ammoniagenes . In mammals and in yeast the conversion of DB00140 ( RF ) into flavin mononucleotide ( Q68DA7 ) and flavin adenine dinucleotide ( DB03147 ) is catalysed by the sequential action of two enzymes : an DB00171 :riboflavin kinase ( Q969G6 ) and an DB00171 : Q8NFF5 ( FMNAT ) . However , most prokaryotes depend on a single bifunctional enzyme , DB03147 synthetase ( FADS ) , which folds into two modules : the C-terminal associated with Q969G6 activity and the N-terminal associated with FMNAT activity . Sequence and structural analysis suggest that the 28-HxGH-31 , 123-Gx(D/N)-125 and 161-xxSSTxxR-168 motifs from FADS must be involved in DB00171 stabilisation for the adenylylation of Q68DA7 , as well as in DB03147 stabilisation for DB03147 phyrophosphorolysis . Mutants were produced at these motifs in the Corynebacterium ammoniagenes FADS ( CaFADS ) . Their effects on the kinetic parameters of CaFADS activities ( Q969G6 , FMNAT and DB03147 pyrophosphorilase ) , and on substrates and product binding properties indicate that H28 , H31 , N125 and S164 contribute to the geometry of the catalytically competent complexes at the FMNAT-module of CaFADS . Transient multiple acyl- DB01992 dehydrogenation deficiency in a newborn female caused by maternal riboflavin deficiency . A newborn female presented on the first day of life with clinical and biochemical findings consistent with multiple acyl- DB01992 dehydrogenase deficiency ( Q8WXG6 ) . DB00140 supplementation corrected the biochemical abnormalities 24 h after commencing the vitamin . In vitro acylcarnitine profiling in intact fibroblasts both in normal and riboflavin depleted media showed normal oxidation of fatty acids excluding defects in electron transfer flavoprotein ( ETF ) , or ETF ubiquinone oxidoreductase ( ETF:QO ) , or a genetic abnormality in flavin metabolism . In addition , sequencing of the genes encoding ETF and ETF:QO in the proband did not reveal any pathogenic mutations . Determination of the maternal riboflavin status after delivery showed that the mother was riboflavin deficient . Repeat testing done two years after the infant 's birth and while on a normal diet showed that the mother was persistently riboflavin deficient and showed a typical Q8WXG6 profile on plasma acylcarnitine testing . A possible genetic defect in riboflavin transport of metabolism in the mother is postulated to be the cause of the transient Q8WXG6 seen in the infant . Sequencing of the Q6ZSM3 , Q969G6 and Q8NFF5 genes encoding key enzymes in riboflavin transport of metabolism in the mother did not identify any pathogenic mutations . The underlying molecular basis of the mother 's defect in riboflavin metabolism remains to be established . Key residues at the riboflavin kinase catalytic site of the bifunctional riboflavin kinase/ Q8NFF5 from Corynebacterium ammoniagenes . Many known prokaryotic organisms depend on a single bifunctional enzyme , encoded by the RibC of RibF gene and named DB03147 synthetase ( FADS ) , to convert DB00140 ( RF ) , first into Q68DA7 and then into DB03147 . The reaction occurs through the sequential action of two activities present on a single polypeptide chain where the N-terminus is responsible for the DB00171 : Q8NFF5 ( FMNAT ) activity and the C-terminus for the DB00171 : riboflavin kinase ( Q969G6 ) activity . Sequence and structural analysis suggest that T208 , N210 and E268 at the C-terminus Q969G6 module of Corynebacterium ammoniagenes FADS ( CaFADS ) might be key during RF phosphorylation . The effect of site-directed mutagenesis on the Q969G6 activity , as well as on substrates and products binding , indicates that T208 and N210 provide the Q969G6 active-site geometry for binding and catalysis , while E268 might be involved in the catalytic step as catalytic base . These data additionally suggest concerted conformational changes at the Q969G6 module of CaFADS during its activity . Mutations at the Q969G6 site also modulate the binding parameters at the FMNAT active site of CaFADS , altering the catalytic efficiency in the transformation of Q68DA7 into DB03147 . This observation supports the hypothesis that the hexameric assembly previously revealed by the crystal structure of CaFADS might play a functional role during catalysis . Q13972 (Mm)/ Q13972 regulates both Ras and Rac signaling pathways . The Q13972 exchange factor molecule contains in addition to the catalytic domain two pleckstrin homology ( P78364 and Q8IXK0 ) , one IQ and one Dbl homology ( DH ) domains . In this study we investigated the role of such additional domains . We found that a Q13972 mutant lacking P78364 and IQ domains is sufficient to activate c-fos promoter in response to lysophosphatidic acid ( P08519 ) . The same mutant did not increase external stimuli-regulated kinase ( P29323 ) activity , suggesting an additional mechanism for the induction of gene transcription . Isolated DH- Q8IXK0 module activates c-Jun NH(2)-terminal kinase and the c-fos promoter in response to P08519 , providing the basis for an P29323 -independent mechanism . These results provide evidence that Q13972 acts as a bifunctional molecule on both P29323 -dependent and independent pathways . DB00160 aminotransferase homologs catalyze the glutamate:glyoxylate aminotransferase reaction in peroxisomes of Arabidopsis . Plant peroxisomal glyoxylate aminotransferases play central roles within the photorespiratory pathway . Genes encoding glyoxylate aminotransferases have been isolated from several animals and microbes , but only recently have plant homologs been identified . Three Arabidopsis homologs of alanine (Ala):glyoxylate aminotransferase 2 ( Q9BYV1 ) contain a putative type 1 peroxisomal targeting signal ( PTS1 ) , but the metabolic significance of these Q9BYV1 homologs is unknown . P19440 and P36268 are Ala aminotransferase ( AlaAT ) homologs from Arabidopsis that represent another type of glyoxylate aminotransferase . These proteins are class I aminotransferases , each containing a putative PTS1 . P19440 and P36268 are members of a small family of AlaATs in Arabidopsis . When expressed as recombinant proteins in Escherichia coli , P19440 and P36268 displayed biochemical characteristics very similar to one another , and to the Arabidopsis protein purified from leaves . Four aminotransferase activities were specifically associated with P19440 and P36268 , using the substrate pairs glutamate ( DB00142 ):glyoxylate , Ala:glyoxylate , DB00142 :pyruvate , and Ala:2-oxoglutarate . P19440 and P36268 may have partially redundant functions ; transcripts of both genes were detected in many of the same tissues . Although DB00142 :glyoxylate aminotransferase ( P19440 ) activity has been observed in several locations in different plants and algae , including the cytoplasm and mitochondria , our subcellular fractionation data indicate that P19440 activity was exclusively peroxisomal in Arabidopsis . Thus , glyoxylate aminotransferase reactions in plant peroxisomes appear to be catalyzed by at least two distinct types of aminotransferases : an AGT1 homolog with serine:glyoxylate aminotransferase activity ( A.H. Liepman , L.J. Olsen [ 2001 ] Plant J 25 : 487-498 ) , and a pair of closely related , potentially redundant AlaAT homologs with P19440 activity . Essential erbB family phosphorylation in osteosarcoma as a target for DB05424 inhibition . BACKGROUND : The role of erbB tyrosine kinases , especially Her-2 , in osteosarcoma has engendered intense debate . Some investigators identified an association between low-level Her-2 expression , compared to none , and poor patient outcome . Others questioned the importance of apparent cytoplasmic expression of Her-2 , since membranous overexpression is associated with poor outcome in carcinomas . We previously demonstrated that primary osteosarcoma cells express cell-surface P00533 and Her-2 , with the p80 isoform of Her-4 localized to the nucleus . We wished to determine if erbB kinases in osteosarcoma were phosphorylated , and if this was required for growth . PROCEDURES : We cultured early passage osteosarcoma cell lines in the presence or absence of the pan-erbB inhibitor DB05424 and examined the phosphorylation status of P00533 , Her-2 , and Her-4 by immunohistochemistry , cell-based ELISA , flow cytometry and two dimensional Western blot . We also assessed the impact of DB05424 upon osteosarcoma growth and survival in vitro . RESULTS : P00533 , Her-2 , and Her-4 were constitutively phosphorylated in early passage osteosarcoma cells cultured in vitro . DB05424 abrogated erbB receptor phosphorylation and caused growth inhibition and apoptosis in a titratible fashion with concentrations of 1 muM or more . CONCLUSIONS : P00533 , Her-2 , and Her-4 are constitutively phosphorylated in early passage osteosarcoma cells in tissue culture , and erbB signaling provides essential growth and anti-apoptotic signals to osteosarcoma cells . This suggests that erbB overexpression is not required for erbB to promote malignancy , but rather that overexpression is one of several mechanisms that generate unregulated erbB signaling . O75365 , a metastasis associated tyrosine phosphatase , is involved in P36888 -ITD signaling and implicated in anti-AML therapy . Combination with other small molecule drugs represents a promising strategy to improve therapeutic efficacy of P36888 inhibitors in the clinic . We demonstrated that combining DB06080 , a P36888 inhibitor , with DB02546 , a HDAC inhibitor , led to synergistic killing of the AML cells with P36888 mutations and suppression of colony formation . We identified a core gene signature that is uniquely induced by the combination treatment in 2 different leukemia cell lines . Among these , we showed that downregulation of O75365 ( O75365 ) played a role in this synergism . O75365 is downstream of P36888 signaling and ectopic expression of O75365 conferred therapeutic resistance through upregulation of P35610 ( signal transducers and activators of transcription ) pathway activity and anti-apoptotic Mcl-1 protein . O75365 interacts with P56524 and DB02546 downregulates O75365 via a proteasome dependent pathway . In addition , O75365 protein was identified in 47 % of AML cases , but was absent in myeloid cells in normal bone marrows . Our results suggest such combination therapies may significantly improve the therapeutic efficacy of P36888 inhibitors . O75365 plays a potential pathological role in AML and it might be a useful therapeutic target in AML , and warrant clinical investigation . Global gene expression profiling and tissue microarray reveal novel candidate genes and down-regulation of the tumor suppressor gene Q03135 in sporadic vestibular schwannomas . BACKGROUND : The vestibular nerve is the predilection site for schwannomas . Few transcriptomic studies have been performed on solely sporadic vestibular schwannomas ( VSs ) . OBJECTIVE : To detect genes with altered expression levels in sporadic VSs . METHODS : We studied 25 VSs and 3 tibial nerves ( controls ) with the ABI 1700 microarray platform . Significance analysis of microarrays was performed to explore differential gene expression . Selected genes were validated with quantitative reverse transcriptase polymerase chain reaction . A tissue microarray was constructed for immunohistochemistry . Neurofibromatosis type II cDNA was sequenced for mutations . RESULTS : The VSs formed 2 clusters based on the total expression of 23,055 genes . Tumor size , previous Gamma Knife surgery , neurofibromatosis type II mutations , and cystic tumors were distributed equally in both . Significance analysis of microarrays detected 1650 differentially expressed genes . On the top 500 list , several cancer-related genes with an unrecognized role in VSs were down-regulated : Q03135 , P10600 , P19320 , P08151 , P10070 , P31323 , P54764 , and Q9UP38 . Immunohistochemistry showed no Q03135 expression in the VSs . The P29323 pathway was the central core in the network linking the differentially expressed genes . The previously reported VS candidate genes P09486 , P00750 , and P05230 were up-regulated . Nineteen of 25 VSs had P35240 mutations . CONCLUSION : Using microarray technology , we identified novel genes and pathways with a putative role in VSs , confirmed previous candidate genes , and found cancer-related genes with no reported role in VSs . Among these , down-regulation of Q03135 at both the mRNA and protein levels is of particular interest because this tumor suppressor normally is expressed in Schwann cells . Partial trypsin digestion as an indicator of mis-folding of mutant alanine:glyoxylate aminotransferase and chaperone effects of specific ligands . Study of a spectrum of missense mutants . DB00160 :glyoxylate aminotransferase ( AGT ) is a liver peroxisomal enzyme whose deficiency results in primary hyperoxaluria type 1 ( P78364 ) . More than 75 P78364 mutations are now documented in the AGT gene ( P21549 ) , of which about 50 % are missense . We have previously demonstrated that many such mutants expressed by transcription/translation are subject to generalized degradation by the proteasome and a specific limited trimming by an endogenous DB00171 -independent protease activity . Here , we report the results of partial digestion using trypsin as a mimic for the endogenous non-proteasomal protease and the use of N-terminal protein sequencing to determine the sensitive site . Partial trypsin digestion also provided an indicator of proper folding of the mutant enzyme . For selected mutations the sensitivity to trypsin could be ameliorated by addition of pyridoxal phosphate or aminooxy acetic acid as specific pharmacological chaperones . Synergism between bosutinib ( DB06616 ) and the Chk1 inhibitor ( PF-00477736 ) in highly imatinib-resistant P11274 /ABL⁺ leukemia cells . Interactions between the dual P11274 / P00519 and Src inhibitor bosutinib and the Chk1 inhibitor PF-00477736 were examined in P11274 / P00519 (+) leukemia cells , particularly imatinib-resistant cells , including those with the T315I mutation . Bosutinib blocked PF-00477736-induced P27361 /2 activation and sharply increased apoptosis in association with Mcl-1 inhibition , p34(cdc2) dephosphorylation , BimEL up-regulation , and DNA damage in imatinib-resistant CML or Ph(+) ALL cell lines . Inhibition of Src or Q02750 by shRNA significantly enhanced PF-0047736 lethality . Bosutinib/PF-00477736 co-treatment also potentiated cell death in P28906 (+) CML patient samples , including dasatinib-resistant blast crisis cells exhibiting both T315I and E355G mutations , but was minimally toxic to normal P28906 (+) cells . Finally , combined in vivo treatment significantly suppressed BaF3/T315I tumor growth and prolonged survival in an allogeneic mouse model . Together , these findings suggest that this targeted combination strategy warrants attention in IM-resistant CML or Ph(+) ALL . Q969G6 couples P01375 receptor 1 to NADPH oxidase . Reactive oxygen species ( ROS ) produced by NADPH oxidase function as defence and signalling molecules related to innate immunity and various cellular responses . The activation of NADPH oxidase in response to plasma membrane receptor activation depends on the phosphorylation of cytoplasmic oxidase subunits , their translocation to membranes and the assembly of all NADPH oxidase components . Tumour necrosis factor ( P01375 ) is a prominent stimulus of ROS production , but the molecular mechanisms by which P01375 activates NADPH oxidase are poorly understood . Here we identify riboflavin kinase ( Q969G6 , formerly known as flavokinase ) as a previously unrecognized P01375 -receptor-1 ( P19438 ) -binding protein that physically and functionally couples P19438 to NADPH oxidase . In mouse and human cells , Q969G6 binds to both the P19438 -death domain and to O75935 (phox) , the common subunit of NADPH oxidase isoforms . Q969G6 -mediated bridging of P19438 and O75935 (phox) is a prerequisite for P01375 -induced but not for Toll-like-receptor-induced ROS production . Exogenous flavin mononucleotide or DB03147 was able to substitute fully for P01375 stimulation of NADPH oxidase in Q969G6 -deficient cells . Q969G6 is rate-limiting in the synthesis of DB03147 , an essential prosthetic group of NADPH oxidase . The results suggest that P01375 , through the activation of Q969G6 , enhances the incorporation of DB03147 in NADPH oxidase enzymes , a critical step for the assembly and activation of NADPH oxidase . DB09559 in the treatment of advanced non-small cell lung cancer : translation from preclinical to clinical development . INTRODUCTION : Treatment outcomes in unselected patients with advanced NSCLC remain disappointing with platinum-based chemotherapy . The addition of monoclonal antibodies targeting P00533 to standard first-line therapy is a validated strategy and has been associated with statistically significant survival advantage in advanced NSCLC . Necitumunab is a fully human IgG1 monoclonal antibody targeting P00533 , having the potential benefit of lower hypersensitivity reaction risk as compared with cetuximab and also equivalent antibody-dependent cell-mediated cytotoxicity . AREAS COVERED : This paper reviews literature on preclinical and early clinical development of necitumumab that is available in PubMed and published abstracts from conferences , as well as ongoing trials as specified by clinicaltrials.gov . Recently , the Phase III clinical trial evaluating the addition of necitumumab to pemetrexed and cisplatin in non-squamous NSCLC was prematurely closed due to concerns about the increased risk of thromboembolic events in the experimental arm . Accrual in the Phase III trial of necitumumab in combination with gemcitabine and cisplatin in squamous NSCLC is ongoing . EXPERT OPINION : Results of the ongoing large randomized trials will be instrumental in determining the drug 's clinical significance and , with the analysis of potential molecular predictive factors , are expected to bring valuable additions to future therapeutic strategies in NSCLC . Imatinib resistance in a novel translocation der(17)t(1;17)(q25;p13) with loss of P04637 but without P11274 / P00519 kinase domain mutation in chronic myelogenous leukemia . We describe here two novel translocations , t(7;14)( O75935 ;q13) and der(17)t(1;17)(q25;p13) , in a 41-year-old man with an accelerated phase ( AP ) of chronic myelogenous leukemia ( CML ) . Chromosome analysis initially showed 46,XY,t(7;14)(p13;q22),t(9;22)(q34;q11.2)[20] . In 3 years , the karyotype evolved to 45,X,-Y,der(7)t(7;14)(p13;q22),t(9;22)(q34;q11.2),-14,der(17)t(1;17)(q25;p13),+der(22)t(9;22)[20] , accompanied with a resistance to imatinib mesylate . The P04637 was deleted from the der(17)t(1;17)(q25;p13) , but there was no mutation of P04637 in the remaining allele . Mutations in the P11274 / P00519 kinase domain could not be detected as well . Morphologically , dysplastic changes including pseudo-Pelger-Huët anomaly appeared in the bone marrow cells . These findings suggest that the t(7;14)( O75935 ;q13) translocation had a crucial role in the progression to CML-AP , and that the resistance to imatinib may be due to the additional cytogenetic abnormalities , including der(17)t(1;17)(q25;p13) , but not to P11274 / P00519 mutations . Discovery and evaluation of 3-phenyl-1H-5-pyrazolylamine-based derivatives as potent , selective and efficacious inhibitors of P07333 -like tyrosine kinase-3 ( P36888 ) . Preclinical investigations and early clinical trial studies suggest that P36888 inhibitors offer a viable therapy for acute myeloid leukemia . However , early clinical data for direct P36888 inhibitors provided only modest results because of the failure to fully inhibit P36888 . We have designed and synthesized a novel class of 3-phenyl-1H-5-pyrazolylamine-derived compounds as P36888 inhibitors which exhibit potent P36888 inhibition and high selectivity toward different receptor tyrosine kinases . The structure-activity relationships led to the discovery of two series of P36888 inhibitors , and some potent compounds within these two series exhibited comparable potency to P36888 inhibitors sorafenib ( 3 ) and DB06080 ( 4 ) in both wt- P36888 enzyme inhibition and P36888 -ITD inhibition on cell growth ( MOLM-13 and MV4;11 cells ) . In particular , the selected compound 12a exhibited the ability to regress tumors in mouse xenograft models using MOLM-13 and MV4;11 cells . Neuroprotective profile of novel P12931 kinase inhibitors in rodent models of cerebral ischemia . Src kinase signaling has been implicated in multiple mechanisms of ischemic injury , including vascular endothelial growth factor ( P15692 ) -mediated vascular permeability that leads to vasogenic edema , a major clinical complication in stroke and brain trauma . Here we report the effects of two novel Src kinase inhibitors , 4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile ( DB06616 ) and 4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[4-(4-methypiperazin-1-yl)but-1-ynyl]-3-quinolinecarbonitrile ( SKS-927 ) , on ischemia-induced brain infarction and short- and long-term neurological deficits . Two well established transient [ transient middle cerebral artery occlusion ( tMCAO ) ] and permanent [ permanent middle cerebral artery occlusion ( pMCAO ) ] focal ischemia models in the rat were used with drug treatments initiated up to 6 h after onset of stroke to mimic the clinical scenario . Brain penetration of Src inhibitors , their effect on blood-brain barrier integrity and P15692 signaling in human endothelial cells were also evaluated . Our results demonstrate that both agents potently block P15692 -mediated signaling in human endothelial cells , penetrate rat brain upon systemic administration , and inhibit postischemic Src activation and vascular leakage . Treatment with DB06616 or SKS-927 ( at the doses of 3-30 mg/kg i.v. ) resulted in a dose-dependent reduction in infarct volume and robust protection from neurological impairments even when the therapy was initiated up to 4- to 6-h after tMCAO . Src blockade after pMCAO resulted in accelerated improvement in recovery from motor , sensory , and reflex deficits during a long-term ( 3 weeks ) testing period poststroke . These data demonstrate that the novel Src kinase inhibitors provide effective treatment against ischemic conditions within a clinically relevant therapeutic window and may constitute a viable therapy for acute stroke . Arthrobacter sp. lipase immobilization for preparation of enantiopure masked beta-amino alcohols . Recent reports on immobilization of lipase from Arthrobacter sp . ( P00519 , MTCC 5125 ; IIIM isolate ) on insoluble polymers have shown altered properties including stability and enantioselectivity . Present work demonstrates a facile method for the preparation of enantiopure beta-amino alcohols by modulation of P00519 enzyme properties via immobilization on insoluble as well as soluble supports using entrapment/covalent binding techniques . Efficacies of immobilized P00519 on insoluble supports prepared from tetraethylorthosilicate/aminopropyltriethoxy silane and soluble supports derived from copolymerization of N-vinyl pyrrolidone-allylglycidyl ether ( P01160 type ) /N-vinyl pyrrolidone-glycidyl methacrylate ( GNP type ) for kinetic resolution of masked beta-amino alcohols have been studied vis-à-vis free P00519 enzyme/wet cell biomass . The immobilized lipase on different insoluble/soluble supports has shown 21-110 mg/g protein binding and 30-700 U/g activity for hydrolyzing tributyrin substrate . The findings have shown a significant enhancement in enantioselectivity ( ee 99 % ) vis-à-vis wet cell biomass providing ee 70-90 % for resolution of beta-amino alcohols . P16284 is involved in P11274 / P00519 signaling and may downregulate imatinib-induced apoptosis of Philadelphia chromosome-positive leukemia cells . P16284 ( CD31 ) is an immunoreceptor tyrosine-based inhibitory motif ( ITIM ) -containing surface glycoprotein expressed on various hematopoietic cells as well as on endothelial cells . P16284 has been shown to play roles in regulation of adhesion , migration and apoptosis . The P11274 / P00519 fusion tyrosine kinase is expressed in chronic myeloid leukemia and Philadelphia-positive ( Ph+ ) acute lymphoblastic leukemia cells , and its inhibition by the clinically used tyrosine kinase inhibitors imatinib or dasatinib induces apoptosis of these cells . In the present study , we demonstrate that P16284 is tyrosine phospho-rylated in its ITIM motifs in various P11274 / P00519 -expressing cells including primary leukemia cells . Studies using imatinib and dasatinib as well as transient expression experiments in 293T cells revealed that P16284 was phosphorylated directly by P11274 / P00519 , which was enhanced by the imatinib-resistant E255K and T315I mutations , or partly by the Src family tyrosine kinases , including Lyn , which were activated dependently or independently on P11274 / P00519 . We also demonstrate by using a substrate trapping mutant of SHP2 that tyrosine phosphorylated P16284 binds SHP2 and is a major substrate for this tyrosine phosphatase in P11274 / P00519 -expressing cells . Overexpression of P16284 in P11274 / P00519 -expressing cells , including K562 human leukemia cells , enhanced cell adhesion and partially inhibited imatinib-induced apoptosis involving mitochondria depolarization and caspase-3 cleavage , at least partly , in an ITIM-independent manner . These data suggest that P16284 may play a role in regulation of apoptosis as well as adhesion of P11274 / P00519 -expressing cells to modulate their imatinib sensitivity and would be a possible candidate for therapeutic target in Ph+ leukemias . Phase 1 study in Japan of siltuximab , an anti- P05231 monoclonal antibody , in relapsed/refractory multiple myeloma . DB09036 , a chimeric monoclonal antibody with high affinity and specificity for interleukin-6 , has been shown to enhance anti-multiple myeloma activity of bortezomib and corticosteroid in vitro . We evaluated the safety , pharmacokinetics , immunogenicity , and antitumor effect of siltuximab in combination with bortezomib and dexamethasone in Japanese patients with relapsed or refractory multiple myeloma . This open-label , phase 1 , dose-escalating study used two doses of siltuximab : 5.5 and 11.0 mg/kg ( administered on day 1 of each 21-day cycle ) . In total , nine patients were treated . The most common grade 3/4 adverse events , lymphopenia ( 89 % ) and thrombocytopenia ( 44 % ) , occurred in patients receiving both doses of siltuximab ; however , no dose-limiting toxicities ( DLTs ) were observed . Following intravenous administration of siltuximab at 5.5 and 11.0 mg/kg , the maximum serum concentration and the area under the curve from 0 to 21 days and from 0 to infinity increased in an approximately dose-proportional manner . Mean half-life , total systemic clearance , and volume of distribution were similar at doses of 5.5 and 11.0 mg/kg . Across both doses , six of the nine patients had complete or partial response ( 22 and 44 % , respectively ) . In conclusion , as no DLT was observed , the recommended dose for this combination is 11.0 mg/kg once every 3 weeks . The study is registered at http://www.clinicaltrials.gov as NCT01309412 .

MH_train_1301

MH_train_1301

MH_train_1301

interacts_with DB00391?

multiple_choice

Detection of thymidylate synthase modulators by a novel screening assay . P04818 ( TS ) , a key cancer chemotherapeutic target , catalyzes the conversion of deoxyuridylate to thymidylate . TS can serve as a repressor of its own synthesis by binding to its own mRNA through TS-specific binding elements ( TBEs ) . In this report , we describe the use of a luciferase reporter plasmid containing two TBEs that can be used as a tool for the identification and initial profiling of compounds that modulate TS activity , levels , or ability to bind mRNA . To validate this model , we evaluated several groups of drugs . Thus , cells were exposed to the pyrimidine analogs 5-fluorouracil ( DB00544 ) , 5-fluorouridine ( DB01629 ) , 5-fluoro-2'-deoxyuridine ( FUdR ) , trifluorothymidine ( DB00432 ) ; to the nonpyrimidine TS-inhibitors AG-331 , nolatrexed ( AG337 ) , and raltitrexed ( DB00293 ) ; or to drugs with other primary sites of action ( methotrexate , actinomycin D , 5-azacytidine , 8-thioguanosine ) . Except for 5-azacytidine and 8-thioguanosine , all compounds examined induced luciferase activity compared with untreated cells . Effects of luciferase activity inducing drugs through TS-affected translation were confirmed by examinations of TS protein and mRNA levels . Treatment of H630- P13671 cells with DB00544 , DB01629 , FUdR , DB00432 , AG331 , AG337 , DB00293 , and methotrexate up-regulated TS levels as determined by Western blot analysis , although TS mRNA levels remained unchanged as determined by reverse transcription-polymerase chain reaction . Our studies demonstrate a novel application of a TBE-dependent reporter plasmid that could be used for the high-throughput identification of potential chemotherapeutic agents that modulate TS RNA-binding activity , either directly or indirectly . Levels of [(3)H]pirenzepine binding in Brodmann 's area 6 from subjects with schizophrenia is not associated with changes in the transcription factor SP1 or P56817 . Decreased muscarinic M1 receptor ( P11229 ) mRNA has been reported in Brodmann 's area ( BA ) 6 from subjects with schizophrenia . We have extended this study by measuring levels of P11229 ( [(3)H]pirenzepine binding ) , P20309 ( [(3)H]4-DAMP binding ) , the transcription factor SP1 and the P11229 downstream target beta-site P05067 -cleaving enzyme 1 ( P56817 ) in BA 6 from 19 subjects with schizophrenia and 19 control subjects . Radioligand binding was quantified using either in situ radioligand binding with autoradiography or , in cohorts of 10 control subjects and 10 subjects with schizophrenia , membrane enriched fraction ( MEF ) CNS ( [(3)H]pirenzepine binding only ) . Levels of SP1 and P56817 were measured by Western blotting . [(3)H]pirenzepine binding to tissue sections was in two layers , binding to tissue sections ( Binding layer 1 : p < 0.01 ; Binding layer 2 : p < 0.001 ) and MEF ( p < 0.05 ) were decreased in schizophrenia . Levels of [(3)H]4-DAMP binding , SP1 and P56817 were not altered in subjects with the disorder . This study shows a decrease in levels of P11229 in BA 6 from subjects with schizophrenia ; as P11229 and BA 6 are important in maintaining normal cognitive function , these data support the hypothesis that decreased levels of cortical P11229 may contribute to the cognitive deficits associated with schizophrenia . Our findings on P56817 suggest that the schizophrenia phenotype reported in P56817 (-/-) mice is not simply due to lack of that protein in the cortex . Reduced satiating effect of d-fenfluramine in serotonin 5-HT(2C) receptor mutant mice . RATIONALE : d- DB00574 stimulates the release of serotonin ( 5-HT ) and is a potent inhibitor of the re-uptake of 5-HT into nerve terminals . Administration of d-fenfluramine suppresses food intake in both animals and humans . OBJECTIVE : We have investigated the role of the P28335 receptor in mediating the effect of d-fenfluramine on mouse food intake and the behavioural satiety sequence . METHODS : Mutant mice lacking serotonin P28335 receptors and wild-type animals were habituated to a daily presentation of wet mash . Animals were non-deprived and received d-fenfluramine ( 3-30 mg/kg ) 30 min prior to being assessed for the presence of stereotypy and presented with wet mash . The behaviour of animals was observed for the subsequent 40 min and food intake was recorded . RESULTS : d- DB00574 dose-dependently inhibited the consumption of a palatable wet mash by the mice . d- DB00574 ( 3 mg/kg ) significantly reduced the amount of wet mash consumed by wild-type mice and induced a temporal advance in the behavioural satiety sequence consistent with an enhancement of satiety . Mutant mice were less sensitive to the satiating effects of 3 mg/kg d-fenfluramine . Hence , this dose of d-fenfluramine had a reduced effect on both food consumption and the behavioural satiety sequence in the P28335 mutant mice . In contrast , mutant mice showed an increased sensitivity to the stereotypy induced by high doses of d-fenfluramine ( 10 , 30 mg/kg ) compared to that of wild-type littermates . CONCLUSION : These data demonstrate a role for the P28335 receptor in mediating d-fenfluramine-induced satiety . Dopamine receptor D2 gene is associated with weight gain in schizophrenic patients under long-term atypical antipsychotic treatment . OBJECTIVE : Schizophrenic patients treated with atypical antipsychotics ( AAPs ) often develop excessive body weight gain ( BWG ) , which may lead to further morbidity and poor treatment compliance . This study examined whether genetic variants in the dopamine receptor D2 ( P14416 ) gene may be associated with body weight change after P08697 treatment . METHODS : The study included 479 schizophrenic patients treated with clozapine ( n=239 ) , olanzapine ( n=70 ) or risperidone ( n=170 ) for an average of 48.2+/-27.8 months . BWG was defined as an increase of more than 7 % of the baseline body weight during P08697 treatment . Thirteen common single nucleotide polymorphisms of the P14416 gene were chosen as tagging single nucleotide polymorphisms . RESULTS : In single-marker-based analysis , the P14416 rs4436578-C homozygous genotype was found to be associated with a significantly increased risk of BWG [ P=0.001 , adjusted odds ratio=3.36 ( 95 % confidence interval=1.62 - 7.00 ) ] . In addition , haplotype analysis further showed that the rs4436578-C-allele-related haplotype was more frequent in those patients with BWG than those without ( P=0.01 - 0.00019 ) . CONCLUSION : Our findings confirm the importance of genetic factors in body weight change induced by long-term P08697 treatment in patients with schizophrenia and indicate a role of P14416 in body weight regulation during long-term P08697 treatment . Antiarthritic mechanisms of amyrin triterpenes . The triterpenes , alpha-amyrin ( AA ) and its palmitate ( P08697 ) and linoleate esters ( AAL ) , were tested on models of inflammatory and destructive arthritic processes and their effects were compared with the clinical antiarthritic drugs indomethacin ( IN ) and methotrexate ( MTX ) . The triterpenes had no effect on the prostaglandin phase of carrageenin pedal edema in rats , which was reduced 28 % by 100 microM IN . AAL caused a considerable reduction in the synthesis by human neutrophils of P09917 products -- 5-HETE ( IC50 = 70 microM ) , LTB4 , ( 62 microM ) , isomer I ( 30 microM ) and isomer II ( 24 microM ) . Rat osteosarcoma cell growth was inhibited by all triterpenes with IC50 's ( microM ) of < 10 ( P08697 ) , 14 ( AA ) and 27 ( AAL ) and were more effective than IN ( 35 ) . MTX caused 100 % inhibition at a concentration of 10 microM compared with 64 % inhibition by P08697 . Tadpole collagenase digestion of type I ( bone ) native collagen was completely inhibited by all the triterpenes as well as IN and MTX at 100 microM . The results indicate that the principal point of antiarthritic intervention by amyrin triterpenes lies in their local inhibition of joint destruction . Contribution of the 37-kDa laminin receptor precursor in the anti-metastatic P08118 -derived peptide DB04985 cell surface binding . PURPOSE : DB04985 is an anti-metastatic synthetic peptide with promising therapeutic efficacy against hormone-refractory prostate cancer . The characterization of the DB04985 peptide cell surface binding/internalization mechanisms and of the receptors involved remained to be explored . RESULTS : [(14)C] DB04985 cell surface binding assays showed rapid and transient kinetic profile , that was inhibited by RGD peptides , laminin , hyaluronan , and type-I collagen . RGD peptides were however unable to inhibit DB04985 intracellular uptake . Far-Western ligand binding studies enabled the identification of the 37-kDa laminin receptor precursor ( P08865 ) as a potential ligand for DB04985 . Overexpression of the recombinant P08865 indeed led to an increase in DB04985 binding but unexpectedly not to its uptake . CONCLUSIONS : Our data support the implication of laminin receptors in cell surface binding and in transducing DB04985 anti-metastatic effects , and provide a rational for targeting cancers that express high levels of such laminin receptors . Genetic influences on outcome following traumatic brain injury . Several genes have been implicated as influencing the outcome following traumatic brain injury ( TBI ) . Currently the most extensively studied gene has been P02649 . P02649 can influence overall and rehabilitation outcome , coma recovery , risk of posttraumatic seizures , as well as cognitive and behavioral functions following TBI . Pathologically , P02649 is associated with increased amyloid deposition , amyloid angiopathy , larger intracranial hematomas and more severe contusional injury . The proposed mechanism by which P02649 affects the clinicopathological consequences of TBI is multifactorial and includes amyloid deposition , disruption of cytoskeletal stability , cholinergic dysfunction , oxidative stress , neuroprotection and central nervous system plasticity in response to injury . Other putative genes have been less extensively studied and require replication of the clinical findings . The P21964 and P14416 genes may influence dopamine dependent cognitive processes such as executive/frontal lobe functions . Inflammation which is a prominent component in the pathophysiological cascade initiated by TBI , is in part is mediated by the interleukin genes , while apoptosis that occurs as a consequence of TBI may be modulated by polymorphisms of the p53 gene . The P12821 gene may affect TBI outcome via mechanisms of cerebral blood flow and/or autoregulation and the O00555 gene may exert an influence via the calcium channel and its effect on delayed cerebral edema . Although several potential genes that may influence outcome following TBI have been identified , future investigations are needed to validate these genetic studies and identify new genes that might influence outcome following TBI . Autoantibodies against four kinds of neurotransmitter receptors in psychiatric disorders . There is a hypothesis that autoimmune abnormalities in neurotransmitter receptors might cause some psychiatric disorders . Using a sensitive radioligand assay , we detected serum autoantibodies to recombinant human muscarinic cholinergic receptor 1 ( P11229 , 34.4 % ) , mu-opioid receptor ( P35372 , 13.1 % ) , P08908 ( P08908 , 7.4 % ) , and dopamine receptor D2 ( P14416 , 4.9 % ) in 122 psychiatric patients . Positive antibodies to P11229 were found in 34.1 % , 34.9 % , 33.3 % , and 9.1 % of patients with schizophrenic disorders ( n=44 ) , mood disorders ( n=63 ) , other psychiatric disorders ( n=15 ) and autoimmune diseases ( n=33 ) , respectively . All three patients with neuroleptic maliganant syndrome had high activities of autoantibodies to P11229 , P35372 , and/or P08908 . Our data suggest that autoimmunity to neurotransmitter receptors might be associated with the induction of psychiatric symptoms and have some relation to neuroleptic malignant syndrome . Enhanced fracture repair by leukotriene antagonism is characterized by increased chondrocyte proliferation and early bone formation : a novel role of the cysteinyl LT-1 receptor . Inflammatory mediators and drugs which affect inflammation can influence the healing of injured tissues . Leukotrienes are potent inflammatory mediators , and similar to prostaglandins , are metabolites of arachidonic acid which can have positive or negative effects on bone and cartilage tissues . Here we tested the hypothesis that blocking the negative regulation of leukotrienes , would lead to enhanced endochondral bone formation during fracture repair . A closed femoral fracture was created in mice . Animals were divided into three groups for treatment with either montelukast sodium , a cysteinyl leukotriene type 1 receptor antagonist ( trade name Singulair ) , zileuton , a P09917 enzyme inhibitor ( trade name DB00744 ) , or carrier alone . The fractures were analyzed using radiographs , quantitative gene expression , histology and histomorphometry , and immunohistochemistry . Both the montelukast sodium group and the zileuton group exhibited enhanced fracture repair when compared with controls . Both treatment groups exhibited increased callous size and earlier bone formation when compared to controls as early as day 7 . Gene expression analysis of treatment groups showed increased markers of chondrocyte proliferation and differentiation , and increased early bone formation markers when compared with controls . Treatment with montelukast sodium directly targeted the cysteinyl leukotriene type 1 receptor , leading to increased chondrocyte proliferation at early time points . These novel findings suggests a potential mechanism by which the cysteinyl leukotriene type 1 receptor acts as a negative regulator of chondrocyte proliferation , with important and previously unrecognized implications for both fracture repair , and in a broader context , systemic chondrocyte growth and differentiation . Clinical development of eniluracil : current status . DB03516 is a potent inactivator of dihydropyrimidine dehydrogenase ( Q12882 ) , which is the first enzyme in the degradative pathway of systemically administered 5-fluorouracil ( DB00544 ) . Two completely oral regimens of eniluracil plus DB00544 are being evaluated in clinical trials : ( 1 ) a chronic schedule with both agents administered P55957 in a 10:1 ratio for 28 days of a 5-week course , and ( 2 ) a 5-day schedule of eniluracil once daily on days 1 through 7 and DB00544 once daily on days 2 through 6 . The clinical development of eniluracil is being pursued in several tumor types , including colorectal cancer , breast cancer , and pancreatic cancer . Response rates achieved in a phase II study of the chronic schedule of oral eniluracil/ DB00544 in patients with colorectal cancer compare favorably with those obtained in trials of intravenous DB00544 and leucovorin , while results from other trials are awaited . Safety analysis for the 28-day schedule has revealed a low incidence of severe toxicities , particularly as compared with standard DB00544 regimens . [ DB00391 in the management of functional dyspepsia and delayed gastric emptying ] . DB00391 is a sulpiride isomer that exerts its prokinetic action through a dual mechanism : 1 ) as a P14416 antagonist and 2 ) as a serotonin 5HT(4) receptor agonist , conferring this drug with a cholinergic effect . At a dosage of 25mg three times daily , levosulpiride accelerates gastric and gallbladder emptying . Clinical trials have shown that this agent is more effective than placebo in reducing the symptoms of dyspepsia , while comparative studies have demonstrated that its effect is similar or superior to that of other dopamine antagonists . The safety profile of levosulpiride is good and the frequency of adverse events is similar to that of other D(2) dopamine antagonists . Therefore , this drug is a useful therapeutic option in the management of patients with functional dyspepsia , as well as in those with delayed gastric emptying . DB09302 inhibits atherosclerosis , improves the plaque morphology , and enhances the effects of a statin . Q8NBP7 ( Q8NBP7 ) inhibition is a potential novel strategy for treatment of CVD . DB09302 is a fully human Q8NBP7 monoclonal antibody in phase 3 clinical development . We evaluated the antiatherogenic potential of alirocumab in P02649 *3Leiden. P11597 mice . Mice received a Western-type diet and were treated with alirocumab ( 3 or 10 mg/kg , weekly subcutaneous dosing ) alone and in combination with atorvastatin ( 3.6 mg/kg/d ) for 18 weeks . DB09302 alone dose-dependently decreased total cholesterol ( -37 % ; -46 % , P < 0.001 ) and TGs ( -36 % ; -39 % , P < 0.001 ) and further decreased cholesterol in combination with atorvastatin ( -48 % ; -58 % , P < 0.001 ) . DB09302 increased hepatic P01130 protein levels but did not affect hepatic cholesterol and TG content . Fecal output of bile acids and neutral sterols was not changed . DB09302 dose-dependently decreased atherosclerotic lesion size ( -71 % ; -88 % , P < 0.001 ) and severity and enhanced these effects when added to atorvastatin ( -89 % ; -98 % , P < 0.001 ) . DB09302 reduced monocyte recruitment and improved the lesion composition by increasing the smooth muscle cell and collagen content and decreasing the macrophage and necrotic core content . DB09302 dose-dependently decreases plasma lipids and , as a result , atherosclerosis development , and it enhances the beneficial effects of atorvastatin in P02649 *3Leiden. P11597 mice . In addition , alirocumab improves plaque morphology . Effects of intracerebroventricular corticotropin-releasing hormone and intravenous morphine on cortisol , prolactin and growth hormone secretion in sheep . It has previously been demonstrated that naloxone and morphine modify the adrenocortical and pituitary responses of sheep to stress . Since P06850 acts within the brain to co-ordinate the stress response , the present experiment was conducted to determine whether morphine has similar effects in sheep given oCRH centrally . Plasma concentrations of cortisol , prolactin and growth hormone were measured in blood samples collected at 10 min intervals from sheep ( N = 5 ) over a 3-hr period . Intravenous injections of saline vehicle or morphine sulphate ( 0.4 mg/kg ) were given after 40 min and intracerebroventricular injections of oCRH ( 0 , 5 or 20 micrograms ) were administered after 60 min . Sustained , dose-related , increases in cortisol were induced by oCRH and , in agreement with findings in stressed sheep , these responses were reduced by pretreatment with morphine . P01236 levels appeared to increase after morphine but oCRH , on its own , did not increase prolactin secretion in this study . There was no change in growth hormone concentrations after oCRH whereas morphine transiently stimulated release . P14416 stimulation of Na+/H+ exchange assessed by quantification of extracellular acidification . A microphysiometer was used to quantify the rate of extracellular acidification by P13671 glioma cells and L fibroblasts expressing recombinant dopamine D2 receptors . The dopamine D2 receptor agonist , quinpirole , accelerated the rate of acidification of the medium by P13671 cells expressing either the short or long form of D2 receptors , D2(415) and D2(444) , but not by wild-type cells that were not transfected with a D2 receptor cDNA . The rate of acidification increased with increasing concentrations of quinpirole up to 100 nM . Inhibition of the response by the dopamine D2 antagonist , spiperone , provided additional evidence that the enhanced extracellular acidification resulted from stimulation of D2 receptors . To test the hypothesis that D2 receptor-stimulated extracellular acidification was due to transport of protons by a Na+/H+ antiporter and reflected intracellular alkalinization , the effect of two inhibitors of Na+/H+ exchange , amiloride and methyl-isobutyl-amiloride , was determined . Both compounds inhibited quinpirole-induced extracellular acidification at concentrations that did not alter D2 receptor-mediated inhibition of adenylylcyclase or radioligand binding to D2 receptors . In addition , quinpirole-induced extracellular acidification was greatly inhibited by removal of sodium from the extracellular medium , confirming the participation of Na+/H+ exchange in the extrusion of acid . Quinpirole ( 100 nM ) also increased the rate of extracellular acidification by L cells expressing D2(415) , LZR1 cells . Treatment with pertussis toxin ( 100 ng/ml for 18 h ) had no effect on the quinpirole-induced acid extrusion by C6D2(415) and LZR1 cells , although the same pertussis toxin treatment regimen completely prevented inhibition of adenylylcyclase . We conclude that recombinant D2 receptors accelerate Na+/H+ exchange in P13671 cells and L fibroblasts by a pathway that does not involve inhibition of adenylylcyclase or pertussis toxin-sensitive G proteins . Fetzima ( levomilnacipran ) , a drug for major depressive disorder as a dual inhibitor for human serotonin transporters and beta-site amyloid precursor protein cleaving enzyme-1 . Pharmacological management of Major Depressive Disorder includes the use of serotonin reuptake inhibitors which targets serotonin transporters ( P31645 ) to increase the synaptic concentrations of serotonin . Beta-site amyloid precursor protein cleaving enzyme-1 ( P56817 -1 ) is responsible for amyloid β plaque formation . Hence it is an interesting target for Alzheimer 's disease ( AD ) therapy . This study describes molecular interactions of a new Food and Drug Administration approved antidepressant drug named ' Fetzima ' with P56817 -1 and P31645 . Fetzima is chemically known as levomilnacipran . The study has explored a possible link between the treatment of Depression and AD . ' Autodock 4.2 ' was used for docking study . The free energy of binding ( ΔG ) values for ' levomilnacipran- P31645 ' interaction and ' levomilnacipran- P56817 ' interaction were found to be -7.47 and -8.25 kcal/mol , respectively . DB08918 was found to interact with S438 , known to be the most important amino acid residue of serotonin binding site of P31645 during ' levomilnacipran- P31645 ' interaction . In the case of ' levomilnacipran- P56817 ' interaction , levomilnacipran interacted with two very crucial aspartic acid residues of P56817 -1 , namely , D32 and D228 . These residues are accountable for the cleavage of amyloid precursor protein and the subsequent formation of amyloid β plaques in AD brain . Hence , Fetzima ( levomilnacipran ) might act as a potent dual inhibitor of P31645 and P56817 -1 and expected to form the basis of a future dual therapy against depression and AD . It is an established fact that development of AD is associated with Major Depressive Disorder . Therefore , the design of new P56817 -1 inhibitors based on antidepressant drug scaffolds would be particularly beneficial . Inhibition of neuronal nitric oxide reduces anxiety-like responses to pair housing . Many psychological disorders are characterized by anxiety and alterations in social interactions . Recent studies demonstrate that the chemical messenger nitric oxide ( NO ) can regulate both anxiety and social behaviours . We tested whether an enzyme that produces NO in the brain , neuronal nitric oxide synthase ( P29475 ) , serves as an interface between social interactions and anxiety-like behaviour . Several investigators have observed that mice increase anxiety-like responses in the elevated plus-maze after pair housing . P29475 gene deletion and DB01997 were used to inhibit the production of neuronal NO . Similar to previous studies , pair housing reduced open arm exploration in the elevated plus-maze . Pair housing also increased corticotropin-releasing hormone ( P06850 ) immunoreactive cells in the paraventricular nucleus ( PVN ) of the hypothalamus . Inhibition of NO production increased open arm exploration in pair-housed mice but decreased open arm exploration in individually housed mice . These results suggest that the effect of P29475 inhibition on anxiety-like responses is context dependent and that behavioural responses to social housing are altered after P29475 inhibition . This research suggests that NO may play an important role in mediating the effect social interactions have on anxiety . Meta-analysis of the safety and tolerability of two dose regimens of buspirone in patients with persistent anxiety . Buspirone is an azapirone with P08908 partial agonist activity which has demonstrated efficacy in the treatment of generalized anxiety disorder , commonly referred to as persistent anxiety . In this meta-analysis report , safety results from two studies comparing buspirone 15 mg twice daily ( P55957 ) with buspirone 10 mg three times daily ( TID ) in patients with persistent anxiety are presented . In the study protocols , qualified patients completed a 7-day placebo lead-in phase and were randomized to receive buspirone 30 mg per day , as either a P55957 or TID regimen , for 6-8 weeks . A total of 289 patients received buspirone 15 mg P55957 ( n = 144 ) or 10 mg TID ( n = 145 ) at 15 sites . The incidence of adverse events was similar between the two treatment groups , except for a significantly greater incidence of palpitations in patients receiving buspirone P55957 ( 5 % ) compared to buspirone TID ( 1 % ) . The most frequently reported adverse events for both buspirone P55957 - and TID-treated patients were dizziness , headache , and nausea . No appreciable differences between treatments were observed for vital signs , physical exam , ECG , or clinical laboratory results . A change to P55957 dosing for buspirone may offer convenience and possibly higher compliance in patients with persistent anxiety without compromising the excellent safety and tolerability profile of the medication . Polymorphisms of dopamine receptor/transporter genes and risk of non-small cell lung cancer . BACKGROUND : The dopaminergic pathway may be of interest in assessing risk of non-small cell lung cancer ( NSCLC ) . Dopamine receptors are expressed in alveolar epithelial cells and human lung tumours , and dopamine inhibits both cell proliferation in vitro and growth of lung tumour xenografts in nude mice . Moreover , dopamine selectively inhibits the vascular permeability and angiogenic activity of vascular endothelial growth factor ( P15692 / P15692 ) . The bioavailability of dopamine is regulated by dopamine receptors D2 ( P14416 ) , D4 ( P21917 ) and dopamine transporter 1 ( Q01959 / Q01959 ) genes . METHODS : We have analysed 10 single nucleotide polymorphisms in P14416 , P21917 and Q01959 / Q01959 genes in relation to lung cancer risk in a case-control study of smoking subjects . The study subjects were 413 healthy individuals from general population and 335 NSCLC cases . Both cases and controls were Caucasians of Norwegian origin . RESULTS : We demonstrate that P14416 polymorphisms -141Cdel , 3208G > T , TaqIB ; P21917 -521C > T and Q01959 / Q01959 -1476T > G are associated with a two- to five-fold increased NSCLC risk . The variant alleles of P14416 1412A > G and 960C > G had protective effects . CONCLUSION : The dopamine receptor/transport gene polymorphisms are associated with the risk of NSCLC among smokers . The data show that the polymorphisms resulting in lower dopamine bioavailability were associated with increased risk of NSCLC . Association of genetic polymorphisms with personality profile in individuals without psychiatric disorders . OBJECTIVE : Population-based twin studies demonstrate that approximately 40-50 % of the variability in personality dimensions results from genetic factors . This study assessed selected polymorphisms in the P21964 Val158Met , P21397 3'VNTR , 5HTTLPR , 102T/C 5- Q13049 , Q01959 3'VNTR and P14416 exon 8 genes and evaluated their association with personality profiles , anxiety levels , and depressiveness in healthy subjects . METHODS : This study included 406 unrelated ( mean age 38.51 years ) , mentally and somatically healthy Caucasian subjects of Polish origin . The prevalence of the gene variants mentioned above and their association with personality profiles , anxiety levels , and depressiveness was assessed using the Temperament and Character Inventory , NEO Five-Factor Inventory , Spielberger 's State-Trait Anxiety Inventory and Beck 's Depression Inventory . RESULTS : The effects of the 5HTTLPR gene on the s/s genotype and empathy ( P06681 ) were lowest in the entire group . The effects of gender , age and the Q13049 gene for the T/T genotype and attachment ( Q7Z3Z2 ) were highest in women . The effects of gender , age and the Q01959 gene on the 9/9 Q01959 genotype , compassion ( C4 ) and cooperativeness ( C ) were lowest in women . The effects of gender , age and the P21964 gene on the DB00134 / DB00134 genotype and neuroticism ( P04626 ) NEO-FFI were also lowest in women . CONCLUSIONS : Our results suggest considerable influence of individual genes on the formation of personality traits . Q07973 as a potential target for cancer therapy . Increasing evidence has accumulated to suggest that vitamin D may reduce the risk of cancer through its biologically active metabolite , DB00136 , which inhibits proliferation and angiogenesis , induces differentiation and apoptosis , and regulates many other cellular functions . Thus , it is plausible to assume that rapid clearance of DB00136 by highly expressed Q07973 could interrupt the normal physiology of cells and might be one cause of cancer initiation and progression . In fact , enhancement of Q07973 expression has been reported in literature for many cancers . Based on these findings , Q07973 -specific inhibitors and vitamin D analogs which are resistant to Q07973 -dependent catabolism might be useful for cancer treatment . Q07973 -specific inhibitor VID400 , which is an azole compound , markedly enhanced and prolonged the antiproliferative activity of DB00136 in the human keratinocytes . Likewise , Q07973 -resistant analogs such as 2α-(3-hydroxypropoxy)- DB00136 ( O2C3 ) and its P06681 -epimer ED-71 ( DB05295 ) , and 19nor- 2α-(3-hydroxypropyl)- DB00136 ( MART-10 ) showed potent biological effects . Our in vivo studies using rats revealed that MART-10 had a low calcemic effect , which is a suitable property as an anticancer drug . Much lower affinity of MART-10 for vitamin D binding protein ( DBP ) as compared with DB00136 may be related to its more potent cellular activities . Based on these results , we conclude that ( 1 ) high affinity for P11473 , ( 2 ) resistance to Q07973 -dependent catabolism , ( 3 ) low affinity for DBP , and ( 4 ) low calcemic effect may be required for designing potent vitamin D analogs for cancer treatment . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . In vitro inhibition of angiogenesis by antibodies directed against the 37kDa/67kDa laminin receptor . The 37kDa/67kDa laminin receptor ( P08865 ) is a central receptor mediating interactions between tumour cells and the basement membrane and is thereby a key player in adhesion and invasion , essential processes in metastatic cancer . To affect continued tumour growth , tumours induce angiogenesis for the constant delivery of nutrients and oxygen . This study aims to determine the blocking effect of the anti- P08865 specific antibody , W3 on the angiogenic potential of HUVE ( human umbilical vein endothelial ) cells . Flow cytometric analysis revealed that 97 % of HUVE cells display cell surface P08865 . An angiogenesis assay was conducted employing HUVE cells seeded on the basement membrane reconstituent Matrigel™ supplemented with the pro-angiogenic factor vascular endothelial growth factor ( P15692 ) . Post 18h incubation at 37°C tubular structures , namely tube lengths were assessed . Treatment of established tubular structures with 100 µg/ml anti- P08865 specific antibody completely blocked angiogenesis . Our findings suggest a central role of the 37kDa/67kDa P08865 in tube formation and recommends anti- P08865 specific antibodies as potential therapeutic tools for treatment of tumour angiogenesis . Dopamine receptors and psychiatric drug treatment . The established antipsychotic drugs act mainly by antagonizing dopamine mediated synaptic transmission in the brain . Increase in the rate of production of dopamine metabolites as well as the firing rate of dopamine-containing neurons can be interpreted as compensatory responses to an interruption of synaptic transmission at dopamine nerve terminals . The demonstration of involvement of limbic and cortical mechanisms in the antipsychotic activity of neuroleptic drugs is far more difficult than the involvement of nigro-striatal and tubero-infundibular mechanisms in the neurological and neuroendocrine effects of these drugs . Application of radioreceptor techniques to dopamine research has supported the findings obtained by other neuropsychopharmacological research techniques , providing more direct evidence of dopamine receptor blockade by neuroleptic drugs . Further research is needed especially in studying the nature of the time-dependent adaptive changes at the receptor sites as well as the differences between the different dopamine projections and neural systems in the brain . The different subtypes of dopamine receptors in the brain , currently called D1 and D2 dopamine receptors , seem to be parallel , although in many respects independently-acting regulatory systems . P14416 -selective antagonists such as sulpiride seem to cause selective D2 receptor up-regulation . P01236 secretion seems to be regulated by D2 dopamine receptors . The exact physiological role of D1 dopamine receptors as well as the clinical consequences of selective D1 antagonism is not known . DB00391 and clozapine are examples of atypical neuroleptic compounds that have quite different profile of action , the former having strong and selective antidopaminergic action , the latter combining a number of non- dopaminergic mechanisms with rather slight effects on dopamine receptors. ( ABSTRACT TRUNCATED AT 250 WORDS )

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interacts_with DB01050?

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DB08626 -induced neprilysin inhibition raises amyloid beta levels in rabbit cortex and cerebrospinal fluid . Studies on the pathogenesis of Alzheimer 's disease ( AD ) suggest overproduction of amyloid beta ( Abeta ) may not be the only pathogenic route to AD . Decreased degradation of Abeta is another possible disease mechanism . P08473 is a neutral endopeptidase that has been proposed to be the major enzyme responsible for Abeta degradation . Studies have reported correlations between Abeta deposition and neprilysin activity in the human brain . This study shows that intracerebroventricular infusion of thiorphan , a neprilysin inhibitor , raises cortical and cerebrospinal fluid ( P04141 ) Abeta concentrations in rabbits . Rabbits treated with thiorphan for 5 days had levels of P04141 and cortical Abeta40 that were 147 and 142 % of the control group , respectively . Results for Abeta42 showed a similar trend . The results indicate that age-related decreases of neprilysin could lead to increased brain concentrations of Abeta , plaque formation , and AD . 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 . Monocytes from type 2 diabetic patients have a pro-inflammatory profile . 1,25-Dihydroxyvitamin D(3) works as anti-inflammatory . The exact factors contributing to the pathogenesis of type 2 diabetes remain elusive . Lately , it was suggested that inflammation and activation of the innate immune system could be linked to type 2 diabetes pathogenesis and also to the development of common diabetic complications , mainly atherosclerosis . The aim of this study was to investigate the role of monocytes in this sub-clinical inflammatory state and test 1,25-dihydroxyvitamin D(3) , the active form of Vitamin D , as an anti-inflammatory agent . For this purpose , monocytes from type 2 diabetic patients were compared to monocytes from healthy controls and type 1 diabetic patients . The expression profile of inflammatory markers in freshly isolated and immune-stimulated monocytes was measured by quantitative real-time RT-PCR . Type 2 diabetic patients showed significantly higher expression levels of P01375 , P05231 , IL-1 , P10145 , P35354 , P05362 and P33681 -1 compared to controls and type 1 diabetic patients . 1,25-Dihydroxyvitamin D(3) was able to down-regulate the expression of P01375 , P05231 , IL-1 , and P10145 , confirming its immunomodulatory properties . From these data we concluded that monocytes from type 2 diabetic patients have a pro-inflammatory profile . In addition , 1,25-dihydroxyvitamin D(3) was able to modulate inflammation in these monocytes . Inhibition of Akt/ P31749 by a P35354 inhibitor induces apoptosis in gastric cancer cells . BACKGROUND/AIM : Inhibition of cyclooxygenase-2 has been proposed to be a potential mechanism for the chemoprevention of gastrointestinal tumors by nonsteroidal anti-inflammatory drugs . This study investigates the mechanisms by which the cyclooxygenase-2 inhibitor SC236 induces apoptosis of gastric cancer cell lines and its downstream signaling pathway . METHODS : Two gastric cancer cell lines , AGS and MKN28 , were treated with SC236 and assessed for cell growth and apoptosis . The involvement of mitogen-activated protein kinase and Akt kinase/protein kinase B ( Akt/ P31749 ) pathways and their downstream signalings were studied in the AGS cell line . RESULTS : SC236 treatment induced apoptosis in gastric cancer cells and caused activation of p38 and stress-activated protein kinase/jun kinase , but down-regulated Akt/ P31749 . The specific p38 inhibitor SB203580 and the dominant-negative stress-activated protein kinase/jun kinase both failed , while the constitutively active form of Akt/ P31749 was able to block SC236-induced apoptosis . SC236-induced apoptosis was coupled with release of cytochrome c and activation of caspases . CONCLUSION : One of the pathways involved in SC-236-induced apoptosis in gastric cancer cells is through downregulation of Akt and then release of cytochrome c . P10275 promotes the migration and invasion of upper urinary tract urothelial carcinoma cells through the upregulation of P14780 and P35354 . Dysregulated androgen receptor ( AR ) signaling is implicated in several types of tumor , including carcinomas of the prostate , breast , liver and bladder . However , the contribution of AR to the progression of upper urinary tract urothelial carcinomas ( UUTUC ) has not been fully investigated . In the present study , we demonstrated that the AR is involved in the metastasis and invasiveness of UUTUC cells . We investigated the role of the AR in UUTUC by using UUTUC-derived BFTC 909 cells . The overexpression of AR promotes the migration and invasion of BFTC 909 cells . Expression of migration/invasion-related genes was increased in BFTC 909 cells overexpressing AR determined by qPCR and western blot analyses . The results showed that AR-enhanced migration and invasion of UUTUC cells are linked to the upregulation of the matrix-degrading enzyme P14780 and cyclooxygenase ( P36551 ) -2 . Subsequently , the blocking of P14780 and P35354 signaling by inhibitors suppressed AR-enhanced cell migration and invasion . The results of the present study provide evidence for the first time of the role of AR in the motility and invasion of UUT cancer cells and support the hypothesis that the AR may play a critical role in the establishment of the invasive phenotype in urothelial neoplasia of UUT . Thus , the AR may also serve as a novel biomarker and potential therapeutic target for UUT cancer . P10275 repression of gonadotropin-releasing hormone gene transcription via enhancer 1 . DB00644 ( DB00644 ) plays a major role in the hypothalamic-pituitary-gonadal ( HPG ) axis , and synthesis and secretion of DB00644 are regulated by gonadal steroid hormones . Disruptions in androgen levels are involved in a number of reproductive defects , including hypogonadotropic hypogonadism and polycystic ovarian syndrome . Androgens down-regulate DB00644 mRNA synthesis in vivo and in vitro via an androgen receptor ( AR ) -dependent mechanism . DB02998 ( R1881 ) , a synthetic AR agonist , represses DB00644 expression through multiple sites in the proximal promoter . In this study , we show AR also represses DB00644 transcription via the major enhancer ( DB00644 -E1 ) . A multimer of the -1800/-1766 region was repressed by R1881 treatment . Mutation of two bases , -1792 and -1791 , resulted in decreased basal activity and a loss of AR-mediated repression . AR bound to the -1796/-1791 sequence in electrophoretic mobility shift assays , indicating a direct interaction with DNA or other transcription factors in this region . We conclude that AR repression of DB00644 -E1 acts via multiple AR-responsive regions , including the site at -1792/-1791 . Serious obstetric complications interact with hypoxia-regulated/vascular-expression genes to influence schizophrenia risk . The etiology of schizophrenia is thought to include both epistasis and gene-environment interactions . We sought to test whether a set of schizophrenia candidate genes regulated by hypoxia or involved in vascular function in the brain ( P31749 , P23560 , O75052 , P36544 , P21964 , Q96EV8 , Q99259 , Q14832 , Q99466 , Q02297 , O43272 , P49798 , P01375 ) interacted with serious obstetric complications to influence risk for schizophrenia . A family-based study of transmission disequilibrium was conducted in 116 trios . Twenty-nine probands had at least one serious obstetric complication ( OC ) using the McNeil-Sjostrom Scale , and many of the OCs reported were associated with the potential for fetal hypoxia . Analyses were conducted using conditional logistic regression and a likelihood ratio test ( LRT ) between nested models was performed to assess significance . Of the 13 genes examined , four ( P31749 ( three SNPs ) , P23560 ( two SNPs ) , Q96EV8 ( one SNP ) and Q14832 ( one SNP ) ) showed significant evidence for gene-by-environment interaction ( LRT P-values ranged from 0.011 to 0.037 ) . Although our sample size was modest and the power to detect interactions was limited , we report significant evidence for genes involved in neurovascular function or regulated by hypoxia interacting with the presence of serious obstetric complications to increase risk for schizophrenia . The cannabinoid O75762 agonist cannabichromene inhibits nitric oxide production in macrophages and ameliorates murine colitis . BACKGROUND AND PURPOSE : The non-psychotropic cannabinoid cannabichromene is known to activate the transient receptor potential ankyrin-type1 ( O75762 ) and to inhibit endocannabinoid inactivation , both of which are involved in inflammatory processes . We examined here the effects of this phytocannabinoid on peritoneal macrophages and its efficacy in an experimental model of colitis . EXPERIMENTAL APPROACH : Murine peritoneal macrophages were activated in vitro by LPS . Nitrite levels were measured using a fluorescent assay ; inducible nitric oxide ( P35228 ) , cyclooxygenase-2 ( P35354 ) and cannabinoid ( P21554 and CB2 ) receptors were analysed by RT-PCR ( and/or Western blot analysis ) ; colitis was induced by dinitrobenzene sulphonic acid ( DNBS ) . Endocannabinoid ( anandamide and 2-arachidonoylglycerol ) , palmitoylethanolamide and oleoylethanolamide levels were measured by liquid chromatography-mass spectrometry . Colonic inflammation was assessed by evaluating the myeloperoxidase activity as well as by histology and immunohistochemistry . KEY RESULTS : LPS caused a significant production of nitrites , associated to up-regulation of anandamide , P35228 , P35354 , P21554 receptors and down-regulation of CB2 receptors mRNA expression . Cannabichromene significantly reduced LPS-stimulated nitrite levels , and its effect was mimicked by cannabinoid receptor and O75762 agonists ( carvacrol and cinnamaldehyde ) and enhanced by P21554 receptor antagonists . LPS-induced anandamide , P35228 , P35354 and cannabinoid receptor changes were not significantly modified by cannabichromene , which , however , increased oleoylethanolamide levels . In vivo , cannabichromene ameliorated DNBS-induced colonic inflammation , as revealed by histology , immunohistochemistry and myeloperoxidase activity . CONCLUSION AND IMPLICATIONS : Cannabichromene exerts anti-inflammatory actions in activated macrophages - with tonic P21554 cannabinoid signalling being negatively coupled to this effect - and ameliorates experimental murine colitis . DB09210 defines a new binding site for allosteric modulators of alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid ( AMPA ) receptors . Glutamate receptors are the most prevalent excitatory neurotransmitter receptors in the vertebrate central nervous system and are important potential drug targets for cognitive enhancement and the treatment of schizophrenia . Allosteric modulators of AMPA receptors promote dimerization by binding to a dimer interface and reducing desensitization and deactivation . The pyrrolidine allosteric modulators , piracetam and aniracetam , were among the first of this class of drugs to be discovered . We have determined the structure of the ligand binding domain of the AMPA receptor subtypes P42262 and P42263 with piracetam and a corresponding structure of P42263 with aniracetam . Both drugs bind to P42262 and P42263 in a very similar manner , suggesting little subunit specificity . However , the binding sites for piracetam and aniracetam differ considerably . DB04599 binds to a symmetrical site at the center of the dimer interface . DB09210 binds to multiple sites along the dimer interface with low occupation , one of which is a unique binding site for potential allosteric modulators . This new site may be of importance in the design of new allosteric regulators . P62937 as a target of DB00515 chemosensitizers . Platinum-based chemotherapeutics are the mainstay of treatment of a range of tumors achieving high response rates but limited in the course of disease by appearance of drug resistance . Tumor cells respond with reduced uptake and increased intracellular inactivation of the drugs , as well as increased DNA repair and general resistance to chemotherapyinduced cell death . DB00515 is known to induce expression of cyclophilins , a group of proteins that have peptidyl-prolyl cis-trans isomerase ( PPIase ) and molecular chaperone activities , as stress response . P62937 ( CypA ) and other members of this family are inhibited by cyclosporin A ( DB00091 ) which sensitized diverse drug-resistant tumor cell lines in vitro to cisplatin . This effect of DB00091 was attributed to metabolic changes , inhibition of DNA repair , enhancement of apoptosis , altered intracellular signal transduction or increased production of reactive oxygen species ( ROS ) , although no definitive explanation was provided so far . Several clinical trials employing cisplatin/carboplatin in combination with DB00091 yielded unsatisfactory results . Since viral replication was found to be dependent on cyclophilins of the host cells , effective new inhibitors , different from DB00091 or with low or absent immunosuppressive activity , are in development or clinical trials . Sanglifehrins are more potent than DB00091 and proved to increase toxicity of cisplatin against hepatocellular cancer cells in vitro . These novel cyclophilin inhibitors may offer new opportunities to achieve reversal of resistance to platinumbased drugs in refractory patients . Responsive cancer patients may be enriched in clinical trials by an identification of the downstream targets of Cyps responsible for chemoresistance . Anti-inflammatory effect of transduced PEP-1-cyclophilin A in Raw264.7 cells and 12-O-tetradecanoylphorbol-13-acetate-induced mice . AIMS : P62937 ( CypA ) is an immunophilin that acts as a receptor for the immunosuppressant drug cyclosporine A ( DB00091 ) . CypA has emerged as a potential drug target for several inflammatory diseases , although the details of its mechanism are unclear . We examined the protective effects of CypA on inflammation in Raw 264.7 cells and animal models . MAIN METHODS : A human CypA gene was fused with a protein transduction domain , PEP-1 peptide , to construct a cell permeable PEP-1-CypA protein . The protein expression level of cyclooxygenase-2 ( P35354 ) and cytokines was detected by Western blot , ELISA and mRNA level of P35354 and cytokines were measured by RT-PCR . The nuclear factor-kappa B ( NF-kB ) and mitogen-activated protein kinase ( MAPK ) activation were analyzed by Western blot and electrophoretic mobility shift assay . Skin inflammation was detected with immunohistochemistry . KEY FINDINGS : Transduced PEP-1-CypA protein markedly inhibited lipopolysaccharide- and 12-O-tetradecanoyl phorbol-13-acetate-induced expression levels of P35354 as well as pro-inflammatory cytokine levels in vitro and in vivo . Furthermore , transduced PEP-1-CypA protein resulted in a significant reduction in the activation of NF-kB and MAPK . SIGNIFICANCE : The results indicate that PEP-1-CypA inhibits inflammatory response cytokines and enzymes by blocking NF-kB and MAPK activation upon stimulation of inflammation in vitro and in vivo . PEP-1-CypA protein may potentially be used as a therapeutic agent against skin diseases-related inflammation . Effects of an alpha 7-nicotinic agonist on default network activity in schizophrenia . BACKGROUND : 3-(2,4-dimethoxybenzylidene)-anabaseine ( DB05708 ) is a partial agonist at α7 nicotinic acetylcholine receptors that has been evaluated clinically for treatment of schizophrenia . This study examined the effects of DB05708 on default network activity as a biomarker for drug effects on pathologic brain function associated with schizophrenia . METHODS : Placebo and two doses of DB05708 were administered in a random , double-blind crossover design during a Phase 2 study of DB05708 . Functional magnetic resonance imaging was performed on 16 nonsmoking patients with schizophrenia while they performed a simple eye movement task . Independent component analysis was used to identify the default network component . Default network changes were evaluated in the context of a polymorphism in P36544 , the α7-nicotinic acetylcholine receptor subunit gene , which was previously found to be associated with schizophrenia . RESULTS : Compared with placebo , both 150 and 75 mg twice daily DB05708 altered default network activity , including a reduction in posterior cingulate , inferior parietal cortex , and medial frontal gyrus activity and an increase in precuneus activity . The most robust difference , posterior cingulate activity reduction , was affected by P36544 genotype . CONCLUSIONS : The observed DB05708 -related changes are consistent with improved default network function in schizophrenia . Pharmacogenetic analysis indicates mediation of the effect through the α7-nicotinic receptor . These results further implicate nicotinic cholinergic dysfunction in the disease and suggest that default network activity may be a useful indicator of biological effects of novel therapeutic agents . P0DMS8 is a critical mediator in LPS-induced pulmonary inflammation . DB00640 receptor A(3) ( A(3) ) regulates directed movement of polymorphonuclear cells ( PMNs ) to sites of inflammation and has been implicated as a relevant mediator in models of inflammatory diseases . Here , we sought to characterize the role of A(3) in a murine model of lung inflammation . Initial studies revealed that pulmonary A(3) transcript levels were elevated following LPS exposure in vivo . In addition , inhalation of LPS increased the accumulation of PMNs in wild-type and A(3)(-/-) mice in all lung compartments . Pretreatment with the specific A(3)-agonist Cl- DB05511 significantly decreased migration of PMNs into lung interstitium and alveolar air space of wild-type mice but not of A(3)(-/-) mice . Lower PMN counts were associated with reduced levels of P01375 -α and P05231 in the alveolar space of wild-type mice that received Cl- DB05511 . In addition , Cl- DB05511 attenuated LPS-induced microvascular permeability in wild-type mice as assessed by the extravasation of Evans blue . In pulmonary microvascular endothelial cells , Cl- DB05511 reduced LPS-induced cytoskeletal remodeling and cell retraction , consistent with a specific role of A(3) for maintaining endothelial integrity . Migratory activity of human PMNs across an endothelial or epithelial monolayer was reduced when A(3) was activated on PMNs . Studies in chimeric mice , however , revealed that Cl- DB05511 required A(3) on both hematopoietic and nonhematopoietic cells to reduce transmigration in vivo . Together , our results shed new light on the role of A(3) in LPS-induced PMN trafficking in the lung and suggest pharmacological modulation of A(3)-dependent pathways as a promising approach in lung inflammation . A DNA vaccine against cytotoxic T-lymphocyte associated antigen-4 ( P16410 ) prevents tumor growth . Co-stimulatory signaling pathway triggered by the binding of P33681 .1/ P33681 .2 ( P33681 /86 ) of antigen-presenting cells ( APCs ) to P10747 of T cells is required for optimal T-cell activation . Cytotoxic T lymphocyte-associated antigen-4 ( P16410 ) is a negative regulator of T cell activation , which competes with P10747 for P33681 .1/ P33681 .2 binding with a greater affinity . DB06186 , a monoclonal antibody against P16410 , has shown positive efficacy in a pivotal clinical trial for the treatment of metastatic melanoma and was approved by FDA . However , the cost of monoclonal antibody-based therapeutics might limit the number of patients treated . To develop a novel therapeutics specifically targeting P16410 , we constructed a DNA vaccine by cloning the sequence of P16410 fused with a transmembrane domain sequence of placental alkaline phosphatase ( PLAP ) into a mammalian expression plasmid , pVAC-1 . Immunization with the resulting construct , pVAC-1-hCTLA-4 , elicited antibody specific to human P16410 with cross reactivity to murine P16410 , which was sufficient for inhibiting B16F10 tumor growth in c57BL/6 mice in the absence of measurable toxicity . Coupling liposome with pVAC-1-mCTLA-4 could break tolerance to self-antigen in BALB/c mice and induce potent immunity against murine P16410 , and suppress growth of subcutaneous renal cell carcinoma ( Renca ) . [ Endocannabinoids -- the new option in the treatment of metabolic syndrome and in smoking cessation ] . Development of the metabolic syndrome results from the interaction of genetic and environmental factors . Metabolic syndrome together with smoking represents risk factors for the development of cardiovascular complications . They may result from the hyperstimulation of the endocannabinoid system . The P21554 receptor has been assumed to play an important role in the endocannabionoid system . It is abundantly expressed in the brain , and in other parts of human body such as in the fat tissue . DB06155 is a selective blocker of cannabinoid-1 ( P21554 ) receptors and participates in the regulation of impaired endocannabinoid system . In the overweight humans , it stimulates sustained reduction of the body weight , girth size and it improves lipid and glucose metabolism . DB06155 also reduces nicotine self-administration and may be effective not only as an aid for smoking cessation but also in the prevention of body weight increase related to the smoking cessation as it was documented in Rio-Lipids and Stratus-us studies . Potential effects of tetrodotoxin exposure to human glial cells postulated using microarray approach . Sodium channels play an important role in many neurological disorders and also in prostate cancer . DB05232 ( TTX ) , a blocker of voltage-gated sodium channels has been chiefly used as a molecular probe for the study and characterization of these channels . The regulation of gene expression in response for the exposure of TTX to glial cells which are reported to be involved in neurodegenerative process is poorly understood . Therefore , the present study aims to develop a repository of genes and map it on a few pivotal neurodegenerative pathways to speculate the effect of TTX . Using Affymetrix GeneChip ( HG-U133A ) , we have selected a subset of 692 differentially expressed genes , several of which are-cullin 4A ( Q13619 ) , ubiquitin carrier protein ( Q16763 ) , proteasome ( prosome , macropain ) subunit , beta type , 8 ( large multifunctional protease 7 ) ( P28062 ) , protein tyrosine phosphatase type IVA ( Q93096 ) , intercellular adhesion molecule 1 ( P05362 ) , prostaglandin-endoperoxide synthase 2 ( P35354 ) , and caspase 1 ( P29466 ) . These genes , which facilitate some of the neurodegenerative pathways , such as ubiquitin , proteasome , inflammation and kinases , were identified to be up- or down-regulated for the TTX treatment . Thus , the selected genes were further examined on ubiquitin-proteasome mediated inflammatory responses pathway as ample evidence for the role of glial cell-mediated inflammation in the neurodegenerative process are available . In summary , our result provides a basic understanding of the differentially expressed genes along with one of the possible pathway which may have been modulated by the exposure of TTX . Novel marine phenazines as potential cancer chemopreventive and anti-inflammatory agents . Two new ( 1 and 2 ) and one known phenazine derivative ( lavanducyanin , 3 ) were isolated and identified from the fermentation broth of a marine-derived Streptomyces sp . ( strain CNS284 ) . In mammalian cell culture studies , compounds 1 , 2 and 3 inhibited P01375 -α-induced NFκB activity ( IC₅₀ values of 4.1 , 24.2 , and 16.3 μM , respectively ) and LPS-induced nitric oxide production ( IC₅₀ values of > 48.6 , 15.1 , and 8.0 μM , respectively ) . PGE₂ production was blocked with greater efficacy ( IC₅₀ values of 7.5 , 0.89 , and 0.63 μM , respectively ) , possibly due to inhibition of cyclooxygenases in addition to the expression of P35354 . Treatment of cultured HL-60 cells led to dose-dependent accumulation in the subG1 compartment of the cell cycle , as a result of apoptosis . These data provide greater insight on the biological potential of phenazine derivatives , and some guidance on how various substituents may alter potential anti-inflammatory and anti-cancer effects . Design and synthesis of an orally bioavailable and selective peptide epoxyketone proteasome inhibitor ( PR-047 ) . Proteasome inhibition has been validated as a therapeutic modality in the treatment of multiple myeloma and non-Hodgkin 's lymphoma . DB08889 , an epoxyketone currently undergoing clinical trials in malignant diseases , is a highly selective inhibitor of the chymotrypsin-like ( CT-L ) activity of the proteasome . A chemistry effort was initiated to discover orally bioavailable analogues of carfilzomib , which would have potential for improved dosing flexibility and patient convenience over intravenously administered agents . The lead compound , 2-Me-5-thiazole- DB00133 (OMe)- DB00133 (OMe)- DB00120 -ketoepoxide ( 58 ) ( PR-047 ) , selectively inhibited CT-L activity of both the constitutive proteasome ( beta5 ) and immunoproteasome ( P28062 ) and demonstrated an absolute bioavailability of up to 39 % in rodents and dogs . It was well tolerated with repeated oral administration at doses resulting in > 80 % proteasome inhibition in most tissues and elicited an antitumor response equivalent to intravenously administered carfilzomib in multiple human tumor xenograft and mouse syngeneic models . The favorable pharmacologic profile supports its further development for the treatment of malignant diseases . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . Tra2betal regulates P19 neuronal differentiation and the splicing of FGF-2R and P42262 minigenes . The present study demonstrates that the expression of Tra2beta1 ( Transformer 2-beta1 ) proteins , an SR ( serine/arginine rich ) protein , is developmentally up-regulated in a neural-specific pattern . The up-regulation is also observed in RA ( retinoic acid ) induced neural differentiation of P19 cells . Tra2betal proteins are located in the nuclei of P19 cells , which are consistent with its functional site as an SR protein . The over-expression of Tra2betal proteins promotes RA induced neuronal differentiation of P19 cells . In P19 cells , the splicing of FGF-2R ( fibroblast growth factor receptor 2 ) minigene produces the P21802 form , while the alternative splicing of P42262 ( glutamate receptor subunit B ) minigene generates two products , the Flop and the Truncated isoforms . Tra2betal inhibits the P21802 splicing , but it promotes the Flop splicing . The results therefore suggest that Tra2betal is involved in the regulation of alternative splicing processes during neural development , peculiarly the splicing of FGF-2R and P42262 genes . Both FGF-2R and P42262 genes are known to play important roles in neural differentiation . Apigenin inhibits release of inflammatory mediators by blocking the NF-κB activation pathways in the HMC-1 cells . Apigenin is a plant flavonoid and a pharmacologically active agent that has been isolated from several plant species . However , the molecular mechanism of apigenin-mediated immune modulation has not been fully understood . One of the possible mechanisms of its protective effects is the down-regulation of inflammatory responses . In this study , we used cells from the human mast cell line ( HMC-1 ) to investigate this effect . Apigenin significantly inhibits the inductive effect of phorbol 12-myristate 13-acetate ( PMA ) plus A23187 on the production of inflammatory cytokines such as tumor necrosis factor ( P01375 ) -α , interleukin ( IL ) -8 , P05231 , and granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) . Moreover , apigenin attenuated the cyclooxygenase ( P36551 ) -2 expression and intracellular Ca(2+) level . In activated HMC-1 cells , apigenin inhibited the PMA plus A23187-induced activation of nuclear factor ( NF ) -κB , IκB degradation , and luciferase activity . Furthermore , apigenin suppressed the expression of P01375 -α , P10145 , P05231 , GM- P04141 , and P35354 by decreasing the intracellular Ca(2+) level and inhibiting NF-κB activation . These results indicate that apigenin has a potential regulatory effect on inflammatory reactions that are mediated by mast cells .

MH_train_1303

MH_train_1303

MH_train_1303

interacts_with DB00741?

multiple_choice

In vitro and in vivo biological activities of a novel nonpolyglutamable anti-folate , MX-68 . MX-68 is a newly synthesized anti-folate , chemically designed not to undergo intracellular polyglutamation and to have increased affinity to dihydrofolate reductase ( P00374 ) . In the present study , we examined the in vitro and in vivo biological activities of MX-68 compared with methotrexate ( MTX ) which forms several polyglutamates intracellularly . MX-68 dose-dependently inhibited the proliferation of PHA- , anti-CD3- , or PMA plus ionomycin-stimulated peripheral blood mononuclear cells ( PBMC ) and endothelial cells ( EC ) from normal subjects as well as P01584 - or P01375 alpha-stimulated synovial fibroblastic cells ( SC ) from rheumatoid arthritis ( RA ) patients . Coaddition of folinic acid completely reversed the anti-proliferative effects of both MX-68 and MTX . Although the anti-proliferative activities of MX-68 were almost comparable to those of MTX , the washout study clearly showed the characteristic nature of MX-68 . When drugs were removed during culture , the suppressive effect of MX-68 completely disappeared , whereas suppression by MTX was merely weakened . MX-68 dramatically suppressed the onset of collagen-induced arthritis ( CIA ) in mice when the drug was orally administered three times a week. starting from the day of first immunization . In this model , 2 mg/kg of MX-68 was sufficient to completely suppress arthritis , whereas suppression by the same dose of MTX was partial . These lines of evidence suggest that polyglutamation is not always a prerequisite in the anti-rheumatic effects of anti-folate . In addition , since intracellular accumulation of polyglutamates is thought to have adverse effects , MX-68 may become a more potent and less toxic anti-rheumatic drug than MTX . The relationship of P04141 and plasma cytokine levels to cerebral white matter injury in the premature newborn . Ischemia and systemic infection are implicated in the etiology of periventricular white matter injury , a major cause of adverse motor and cognitive outcome in preterm infants . Cytokines are signaling proteins that can be produced as part of the inflammatory response to both ischemia and infection . The aim of this study was to relate cerebrospinal fluid ( P04141 ) concentrations of P05231 , P10145 , P22301 , tumor necrosis factor alpha ( P01375 ) , and interferon gamma ( P01579 ) to magnetic resonance-defined white matter injury in preterm infants . Relationships between P04141 and plasma cytokine concentrations were also examined . Preterm infants ( < or=32 wk ) and more mature infants from The Royal Women 's Hospital , Melbourne , Australia , and Christchurch Women 's Hospital , Christchurch , New Zealand , were eligible for study if they required a clinically indicated lumbar puncture . Plasma samples were obtained in a subgroup of Christchurch infants . Preterm infants underwent advanced quantitative volumetric magnetic resonance imaging using a 1.5-Tesla scanner at term equivalent . One hundred forty-six infants were enrolled and 190 P04141 and 42 plasma samples obtained . There was no significant correlation between paired P04141 and plasma concentrations for any cytokine . In comparing plasma and P04141 concentrations , levels of P10145 were significantly higher in P04141 than plasma . Preterm infants with Q9BWK5 -defined cerebral white matter injury had higher levels of P05231 , P22301 , and P01375 in the P04141 than infants without such injury . Plasma cytokine concentrations may not reflect P04141 cytokine levels or inflammatory events within the brain . Elevated P04141 levels of cytokines in infants with white matter injury suggest an altered inflammatory balance . Steatohepatitis in laboratory opossums exhibiting a high lipemic response to dietary cholesterol and fat . Plasma VLDL and LDL cholesterol were markedly elevated ( > 40-fold ) in high-responding opossums , but moderately elevated ( 6-fold ) in low-responding opossums after they had consumed a high-cholesterol and high-fat diet for 24 wk . In both high- and low-responding opossums , plasma triglycerides were slightly elevated , threefold and twofold , respectively . Dietary challenge also induced fatty livers in high responders , but not in low responders . We studied the lipid composition , histopathological features , and gene expression patterns of the fatty livers . Free cholesterol ( 2-fold ) , esterified cholesterol ( 11-fold ) , and triglycerides ( 2-fold ) were higher in the livers of high responders than those in low responders , whereas free fatty acid levels were similar . The fatty livers of high responders showed extensive lobular disarray by histology . Inflammatory cells and ballooned hepatocytes were also present , as were perisinusoidal fibrosis and ductular proliferation . In contrast , liver histology was normal in low responders . Hepatic gene expression revealed differences associated with the development of steatohepatitis in high responders . The accumulation of hepatic cholesterol was concomitant with upregulation of the P04035 gene and downregulation of the Q02318 , Q9H221 , and P21439 genes . Genes involved in inflammation ( P01375 , P19838 , and P35354 ) and in oxidative stress ( P13498 and P14598 ) were upregulated . Upregulation of the growth factor genes ( PDGF and P01137 ) and collagen genes ( Col1A1 , Col3A1 , and Col4A1 ) was consistent with fibrosis . Some of the histological characteristics of the fatty livers of high-responding opossums imitate those in the livers of humans with nonalcoholic steatohepatitis . Molecular pathways : targeting mechanisms of asbestos and erionite carcinogenesis in mesothelioma . Malignant mesothelioma is an aggressive malignancy related to asbestos and erionite exposure . AP-1 transcriptional activity and the NF-κB signaling pathway have been linked to mesothelial cell transformation and tumor progression . P14210 and c- DB00134 are highly expressed in mesotheliomas . Phosphoinositide 3-kinase , AKT , and the downstream P42345 are involved in cell growth and survival , and they are often found to be activated in mesothelioma . p16(INK4a) and p14( Q8N726 ) are frequently inactivated in human mesothelioma , and ∼50 % of mesotheliomas contain the P35240 mutation . Molecular therapies aimed at interfering with these pathways have not improved the dismal prognosis of mesothelioma , except possibly for a small subset of patients who benefit from certain therapies . Recent studies have shown the importance of asbestos-induced inflammation in the initiation and growth of mesothelioma , and P09429 and Nalp3 inflammasome have been identified as key initiators of this process . Asbestos induces cell necrosis , causing the release of P09429 , which in turn may activate Nalp3 inflammasome , a process that is enhanced by asbestos-induced production of reactive oxygen species . P09429 and Nalp3 induce proinflammatory responses and lead to interleukin-1β and P01375 -α secretion and NF-κB activity , thereby promoting cell survival and tumor growth . Novel strategies that interfere with asbestos- and erionite-mediated inflammation might prevent or delay the onset of mesothelioma in high-risk cohorts , including genetically predisposed individuals , and/or inhibit tumor growth . The very recent discovery that germline BAP1 mutations cause a new cancer syndrome characterized by mesothelioma , uveal melanoma , and melanocytic tumors provides researchers with a novel target for prevention and early detection . DB04958 in the therapy of oncological and immunological diseases . B cells play an important role in the pathogenesis of certain lymphomas and leukemias , as well as many autoimmune diseases . Antagonistic B-cell antibodies are thus gaining an increasing role in the management of these diseases . The first antibody target in this regard was P11836 , with the development and introduction of rituximab in the management of B-cell malignancies , as well as rheumatoid arthritis . A second candidate target is P20273 . The first antagonistic antibody to this B-cell marker , epratuzumab , appears to function , in contrast to P11836 antibodies , more by modulation of B cells rather than by their high depletion in circulation . Originally developed for the treatment of non-Hodgkin 's lymphoma , epratuzumab has now been found to be effective , with a very good safety profile , in two prototype autoimmune diseases : systemic lupus erythematosus and primary Sjögren 's syndrome . Recent studies have demonstrated the activity and safety of epratuzumab in non-Hodgkin 's lymphoma patients who have relapsed or are refractive to conventional therapy , including rituximab , and has also shown good activity in follicular and diffuse large B-cell lymphoma in combination with rituximab . As such , this new investigative antibody may have a significant market potential owing to the multitude of diseases and patients who may benefit from a P20273 , B-cell antibody immunotherapy that is complementary to the known effects and role of P11836 antibodies , but can usually be administered within 1 h and depletes approximately 50 % of circulating B cells . Monoclonal antibodies targeting P01584 reduce biomarkers of atherosclerosis in vitro and inhibit atherosclerotic plaque formation in P02649 -deficient mice . OBJECTIVE : Atherosclerosis is a condition that is increasingly contributing to worldwide mortality through complications such as stroke and myocardial infarction . IL-1β plays multiple direct , local roles in the formation and stability of the atheroma by eliciting the production of additional cytokines and proteolytic enzymes from macrophages , endothelial cells ( EC ) and smooth muscle cells ( SMC ) . We therefore tested whether an anti-IL-1β antibody , DB06062 , might inhibit the secretion of pro-atherogenic cytokines from macrophages in vitro and affect a positive outcome in the P02649 -deficient mouse ( ApoE(-/-) ) model of atherosclerosis in vivo . METHODS AND RESULTS : In an in vitro co-culture model , DB06062 inhibited macrophage-induced secretion of key atherogenic cytokines from EC and SMC , including P05231 , P10145 , P13500 and TNFα . The release of degradative enzymes , such as the matrix metalloproteinases P08254 and P14780 , was also decreased by DB06062 . In addition , DB06062 inhibited the secretion of P13232 from EC and P05112 from SMC , cytokines not previously reported to be driven by IL-1β in this context . In vivo , XMA052 MG1K , a chimeric murine version of DB06062 , inhibited the formation of atherosclerotic lesions in the ApoE(-/-) model at all three doses tested . This effect was comparable to that reported for complete genetic ablation of IL-1β or IL-1R1 on an ApoE(-/-) background and was associated with decreases in plasma non-HDL/HDL cholesterol ratio and plaque lipid content and macrophage infiltration . CONCLUSIONS : These results demonstrate for the first time that an antibody targeting IL-1β can inhibit the progression of atherosclerosis in vivo , highlighting the importance of this key cytokine in cardiovascular disease . P04150 -mediated regulation of P14780 gene expression in human ovarian surface epithelial cells . OBJECTIVE : To obtain proof-of-concept that locally produced anti-inflammatory steroids suppress ovulation-associated extracellular matrix proteases in human ovarian surface epithelial ( OSE ) cells . DESIGN : Primary OSE cell cultures treated with interleukin-1alpha ( IL-1alpha ) ( 500 pg/mL ) as proxy for inflammation , with/without anti-inflammatory steroid ( cortisol or progesterone [ P ] , 0.01-1.0 microM ) . SETTING : Academic medical center . PATIENT(S) : Sixteen premenopausal women ( 29-46 years ) undergoing surgery for nonmalignant gynecological conditions . MAIN OUTCOME MEASURE(S) : Semiquantitative extracellular matrix protease gene expression profiling with verification by real-time quantitative reverse transcription polymerase chain reaction ( qRT-PCR ) and gelatinase zymography . RESULT(S) : Treatment with IL-1alpha stimulated messenger RNA ( mRNA ) expression of several ovulation-associated matrix metalloproteinase genes by OSE cell cultures , including gelatinase B ( P14780 ) but not gelatinase A ( P08253 ) . The IL-1alpha-stimulated P14780 mRNA production was suppressed by cortisol but not P. DB00741 but not P also dose-dependently suppressed IL-1alpha-stimulated P14780 gelatinase activity and this effect was blocked by the glucocorticoid receptor antagonist DB00834 . CONCLUSION(S) : In human OSE cells , stimulation of P14780 gene expression and proteolytic activity by IL-1alpha is suppressed by anti-inflammatory cortisol through a glucocorticoid receptor-mediated mechanism . Because IL-1alpha also generates cortisol formation in OSE by stimulating cortisone reductase activity , these results support a role for intracrine cortisol in minimizing proteolytic damage to the OSE at ovulation . The effects of different types of statins on proliferation and migration of P14210 -induced Human Umbilical Vein Endothelial Cells ( HUVECs ) . BACKGROUND AND AIM : Statins are P04035 inhibitors within the framework of cholesterol biosynthesis and used to lower the low-density lipoprotein ( LDL ) . There are other aspects of statins can deploy a protective effect , even without the LDL 's lowering . The aim of this study is to investigate the effects of different type of statins on proliferative and migrative behaviors of Hepatocyte Growth Factor ( P14210 ) induced human umbilical vein endothelial cells ( HUVECs ) . MATERIALS AND METHODS : Human umbilical vein endothelial cells were isolated and cultured . Groups were designed in order to observe the effects of every individual substance . HUVECs were stimulated with P14210 , statins and farnesylpyrophosphat ammonium salt ( FPP ) or geranylgeranyl-pyrophosphate ( GGPP ) , respectively . Cell proliferations were counted 48 hours after initial stimuli and distances between migration fronts were used in migration analyses . RESULTS : All types of statins showed significant anti-migrative and anti-proliferative characters . Simvastatin and fluvastatin but not cerivastatin , were able to inhibit the P14210 -depending migration and showed a significant effect on the inhibition of the isoprenylation ( GGPP ) . Only simvastatin influenced the P14210 -depending migration via inhibiting the isoprenylation process through GGPP . DB00439 significantly decreased the proliferation and DB01095 significantly enhanced the migration behaviors of HUVECs when they were co-incubated with methyl-8-cyclodextrin ( O95822 ) . CONCLUSIONS : Statins countermand the proproliferative and as well as the promigrative effect of P14210 on HUVECs . The mechanisms which provoke this effect are dependent on the type of statin . Direct interactions of statins with lipid rafts play a significant role in the endothelial cell mechanisms . Donor pre-treatment with everolimus or cyclosporine does not reduce ischaemia-reperfusion injury in a rat kidney transplant model . BACKGROUND : Immunosuppressive agents have been investigated in renal ischaemia-reperfusion injury ( IRI ) and have frequently demonstrated a beneficial effect . Most studies focused on treatment of the recipient at the time of transplantation . Pre-treatment of these organs before injury ( pharmacological pre-conditioning ) may particularly protect these organs . This study aimed to investigate the possible protective effects of donor pre-treatment with cyclosporine ( DB00091 ) or the P42345 inhibitor everolimus or their combination against IRI during renal transplantation in a rat model . METHODS : Donors received vehicle , DB00091 ( 5 mg/kg ) , everolimus ( 0.5 mg/kg ) or CsA + everolimus . Two oral doses were administered to the donors at 24 h and again at 6 h prior to donor kidney removal . Syngeneic rat kidneys were preserved in UW solution for 24 h prior to transplantation . After 24 h of reperfusion , blood and tissue samples were collected from recipients for further analysis . RESULTS : Renal functions as determined by creatinine and necrosis scores were not different between the experimental groups . Cleaved caspase-3 , heat shock protein 70 ( HSP70 ) , tumor-necrosis factor-alpha ( P01375 -α ) and nitrotyrosine protein levels were not statistically different between the four treatment groups at 24 h post-transplantation . Blood NMR analysis on metabolic markers for IRI reveals no beneficial effects of donor pre-treatment on the 24-h outcome in transplantation . CONCLUSIONS : When given alone or as a combination to donors before organ recovery , cyclosporine or everolimus does not appear to ameliorate IRI . Changes of thyroid hormone levels and related gene expression in zebrafish on early life stage exposure to triadimefon . In this study , zebrafish was exposed to triadimefon . Thyroid hormones levels and the expression of related genes in the hypothalamic-pituitary-thyroid ( Q9HD23 ) axis , including thyroid-stimulating hormone ( P01222 ) , deiodinases ( dio1 and dio2 ) and the thyroid hormone receptor ( thraa and thrb ) were evaluated . After triadimefon exposure , increased DB00451 can be explained by increased thyroid-stimulating hormone ( P01222 ) . The conversion of DB00451 to DB00279 ( deiodinase type I-dio1 ) was decreased , which reduced the DB00279 level . P10828 ( thrb ) mRNA levels were significantly down-regulated , possibly as a response to the decreased DB00279 levels . The overall results indicated that triadimefon exposure could alter gene expression in the Q9HD23 axis and that mechanisms of disruption of thyroid status by triadimefon could occur at several steps in the synthesis , regulation , and action of thyroid hormones . Effect of N9-methylation and bridge atom variation on the activity of 5-substituted 2,4-diaminopyrrolo[2,3-d]pyrimidines against dihydrofolate reductases from Pneumocystis carinii and Toxoplasma gondii . The effect of N9-methylation and bridge atom variation on inhibitory potency and selectivity of 2,4-diaminopyrrolo[2,3-d]pyrimidines against dihydrofolate reductases ( P00374 ) was studied . Specifically three nonclassical 2,4-diamino-5-((N-methylanilino)methyl)pyrrolo[2,3-d]pyrimidines with 2',5'-dimethoxyphenyl ( 2 ) , 3',4'-dichlorophenyl ( 3 ) , 1'-naphthyl ( 4 ) , one classical analogue with a 4'-L-glutamate substituent ( 10 ) , and four nonclassical 2,4-diamino-5-((phenylthio)methyl)pyrrolo[2,3-d]pyrimidines with 3',4'-dimethoxyphenyl ( 5 ) , 3',4'-dichlorophenyl ( 6 ) , 1'-naphthyl ( 7 ) , and 2'-naphthyl ( 8 ) substituents were synthesized . The classical and nonclassical analogues were obtained by displacement of the intermediate 2,4-diamino-5-bromomethylpyrrolo[2,3-d]pyrimidine , 14 , with appropriately substituted N-methylaniline , thiophenols , or 4-(N-methylamino)benzoyl-L-glutamate . Compounds 2-8 and 10 were evaluated against Pneumocystis carinii ( pc ) , Toxoplasma gondii ( tg ) , and rat liver ( rl ) DHFRs . The N-methyl and thiomethyl analogues were more inhibitory than their corresponding anilinomethyl analogues ( previously reported ) against all three DHFRs . The inhibitory potency of these analogues was greater against rlDHFR than against tgDHFR which resulted in a loss of selectivity for tgDHFR compared to the N9-H analogues . The classical N9-methyl analogue 10 was more potent and about 2-fold more selective against tgDHFR than its corresponding desmethyl analogue . All of the analogues , 2-8 and 10 , were more selective than trimetrexate ( DB01157 ) against pcDHFR ( except 4 ) and significantly more selective than DB01157 against tgDHFR . Molecular characteristics of the N-terminal region of the quail follitropin receptor . We determined the primary structure of follitropin receptor ( P23945 ) at its N-terminal extracellular domain , which is the key region of specific hormone binding in avian ( quail ) species . In this region , quail P23945 showed about 70 % homology with mammalian DB00094 -Rs at the level of predicted amino acid sequences . The leucine-rich repetitive motif which is conserved in all mammalian DB00094 -Rs was also detected in the quail P23945 . However , some unique amino acid replacements were found at the positions of cystein residues and potential N-linked glycosylation sites . The sequence of the quail lutropin receptor ( LH-R ) previously defined by us showed a homology between P23945 and LH-R of 47.4 % . This value is close to those between mammalian DB00094 -Rs and LH-Rs , which in the corresponding region are human , 50.3 % ; rat , 50.9 % . A pharmacokinetic and clinical assessment of tofacitinib for the treatment of rheumatoid arthritis . INTRODUCTION : Rheumatoid arthritis ( RA ) is a chronic painful and debilitating autoimmune disease . Although the outcome for patients with RA has improved markedly in the past decades , driven largely by the advent of biological disease-modifying antirheumatic drugs ( DMARDs ) and updated management strategies , adequate disease control can not be achieved in a substantial proportion of patients . Since RA is a syndrome with different biological subsets , DMARDs , with a novel mechanism of action , may represent a valuable addition to the current armamentarium . DB08895 is a novel synthetic DMARD that selectively inhibits Janus kinases ( JAKs ) , particularly P23458 and P52333 . AREAS COVERED : This review describes the pharmacokinetics of tofacitinib . Furthermore , the article summarizes and comments the drug 's efficacy and safety profile in RA patients . The authors furthermore assess data derived from the FDA 's RA development program . EXPERT OPINION : DB08895 is an oral synthetic DMARD displaying linear pharmacokinetics . Metabolism , primarily mediated by P08684 , accounts for 70 % of the total clearance of the drug ; the remaining 30 % are renally excreted . DB08895 monotherapy , or in combination with traditional DMARDs , has demonstrated its efficacy while having an acceptable safety profile in RA patients who have responded inadequately to current DMARDs , including P01375 antagonists . In view of its undetermined benefit to risk ratio , in the real-world population , tofacitinib should , for now , only be prescribed to selected patients . The cloning and reintroduction into animal cells of a functional CAD gene , a dominant amplifiable genetic marker . Rodent cells resistant to DB03459 , a specific inhibitor of the aspartate transcarbamylase activity of the multifunctional P27708 , overproduce CAD as a result of amplification of the CAD gene . We cloned a functional CAD gene from Syrian hamster cells using a cosmid vector . Two independently isolated cosmids containing CAD genes have inserts 40 and 45 kb long . We introduced the cloned genes into CAD-deficient Chinese hamster ovary ( CHO ) cell mutants by fusing them with protoplasts of Escherichia coli containing the cosmids . We also introduced the cloned genes into wild-type CHO cells by selecting cells that became resistant to high concentrations of DB03459 following protoplast fusion . The transformants of the mutant and wild-type CHO cells contain multiple active copies of the donated Syrian hamster CAD genes . The cloned genes in three independent transformants are integrated into host-cell chromosomes at single locations identified by in situ hybridization . In two of these transformants , the genes are located in one X chromosome or in a chromosome resembling the X . In the third case , the genes are located in a small metacentric or rearranged chromosome . Monoclonal antibody therapy in haematological malignancies . This review focuses on the development of monoclonal antibodies that have been or are being introduced in the treatment of both lymphoid and myeloid haematological malignancies in adults . After a general introduction on the principles of antibody selection for therapy , this review summarizes the results of the clinical trials that led to the approval of antibodies by the FDA ( Food and Drug administration , USA ) and/or the EMEA ( European Medicines Agency ) , e.g. different anti- P11836 , anti- P31358 and anti- P20138 antibodies . Furthermore , several antibodies that seem promising in phase I/II studies ( like anti- P20273 , anti-CD23 , anti- P01730 , anti- P28908 , anti- P33681 , anti- P15692 , anti HLA-DR and anti- P01375 ) are highlighted . The application of monoclonal antibodies has nowadays become indispensable in the treatment of lymphoma and leukaemia 's and the number of new indications is still growing . Therefore , also some interesting phase III studies that are recruiting patients at this moment , and some new technical developments are discussed . Osteoclastic differentiation and function regulated by old and new pathways . The osteoclast is a specialized multinucleated variant of the macrophage family . It degrades mineralized tissue , and is required for modeling and remodeling of bone . The osteoclast has long been known to require vitamin D for its differentiation and to be regulated by parathyroid hormone via circulating Ca(2+) levels . Two local signals important in osteoclast survival and differentiation , P09603 and O14788 , were characterized by the mid-1990 s . A basic framework of specialized cell attachment and resorption molecules was also clear by that time , including the alpha(v)beta(3) integrin , the key adhesion molecule of the mature osteoclast , the highly expressed vacuolar-type H(+)-ATPase that drives acid secretion to dissolve mineral , and cathepsin K , the predominant acid proteinase for collagenolysis . Recently , additional detail has been added to this framework , showing that the osteoclast has more complex regulation than was previously believed . These include the findings that one component of the V-H(+)-ATPase is unique to the osteoclast , that chloride transport and probably Cl(-)/H(+) exchange are also required for mineral degradation , and that additional receptors besides Q9Y6Q6 and Fms regulate osteoclast formation and survival . Additional receptors include estrogen receptor-alpha , P01375 -family receptors other than Q9Y6Q6 , and , at least in some cases , glycoprotein hormone receptors including the P16473 and the P23945 . Challenges in understanding osteoclast biology include how the signalling mechanisms function cooperatively . Recent findings suggest that there is a network of cytoplasmic adapters , including Gab-2 and P56945 , which are modified by multiple signalling mechanisms and which serve to integrate the signalling pathways . Purification and characterization of mouse O15528 overproduced by an Escherichia coli system coexpressing molecular chaperonins GroEL/ES . The expression of mouse O15528 in Escherichia coli has been dramatically enhanced by coexpression of GroEL/ES . To reveal the enzymatic properties of O15528 , we measured its hydroxylation activity toward vitamin D3 and DB01436 ( 1alpha(OH)D3 ) in addition to the physiological substrate DB00146 . Surprisingly , O15528 converted vitamin D3 to 1alpha, DB00146 . Both 1alpha-hydroxylation activity toward vitamin D3 , and 25-hydroxylation activity toward 1alpha(OH)D3 were observed . The Km and Vmax values for 25-hydroxylation activity toward 1alpha(OH)D3 were estimated to be 1.7 microM and 0.51 mol/min/mol P450 , respectively , while those for 1alpha-hydroxylation activity toward DB00146 were 0.050 microM and 2.73 mol/min/mol P450 , respectively . Note that the substrate must be fixed in the opposite direction in the substrate-binding pocket of O15528 between 1alpha-hydroxylation and 25-hydroxylation . Based on these results and the fact that human Q02318 and Streptomyces CYP105A1 also convert vitamin D3 to 1alpha, DB00146 , 1alpha-hydroxylation , and 25-hydroxylation of vitamin D3 appear to be closely linked together . Polymorphism identification in the P11310 , P01008 , P22301 , P15173 and P01222 genes of cattle . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells . Rheb protein binds CAD ( carbamoyl-phosphate synthetase 2 , aspartate transcarbamoylase , and dihydroorotase ) protein in a GTP- and effector domain-dependent manner and influences its cellular localization and carbamoyl-phosphate synthetase ( CPSase ) activity . Rheb small GTPases , which consist of Rheb1 and Q8TAI7 ( also known as RhebL1 ) in mammalian cells , are unique members of the Ras superfamily and play central roles in regulating protein synthesis and cell growth by activating P42345 . To gain further insight into the function of Rheb , we carried out a search for Rheb-binding proteins and found that Rheb binds to P27708 ( carbamoyl-phosphate synthetase 2 , aspartate transcarbamoylase , and dihydroorotase ) , a multifunctional enzyme required for the de novo synthesis of pyrimidine nucleotides . CAD binding is more pronounced with Q8TAI7 than with Rheb1 . Rheb binds CAD in a GTP- and effector domain-dependent manner . The region of CAD where Rheb binds is located at the C-terminal region of the carbamoyl-phosphate synthetase domain and not in the dihydroorotase and aspartate transcarbamoylase domains . Rheb stimulated carbamoyl-phosphate synthetase activity of CAD in vitro . In addition , an elevated level of intracellular UTP pyrimidine nucleotide was observed in Tsc2-deficient cells , which was attenuated by knocking down of Rheb . Immunostaining analysis showed that expression of Rheb leads to increased accumulation of CAD on lysosomes . Both a farnesyltransferase inhibitor that blocks membrane association of Rheb and knockdown of Rheb mislocalized CAD . These results establish CAD as a downstream effector of Rheb and suggest a possible role of Rheb in regulating de novo pyrimidine nucleotide synthesis .

MH_train_1304

MH_train_1304

MH_train_1304

interacts_with DB00015?

multiple_choice

DB05258 /beta promotes cell survival by activating nuclear factor kappa B through phosphatidylinositol 3-kinase and Akt . Interferons ( IFNs ) play critical roles in host defense by modulating gene expression via activation of signal transducer and activator of transcription ( P35610 ) factors . IFN-alpha/beta also activates another transcription factor , nuclear factor kappaB ( NF-kappaB ) , which protects cells against apoptotic stimuli . NF-kappaB activation requires the IFN-dependent association of P40763 with the P17181 chain of the IFN receptor . IFN-dependent NF-kappaB activation involves the sequential activation of a serine kinase cascade involving phosphatidylinositol 3-kinase ( PI-3K ) and Akt . Whereas constitutively active PI-3K and Akt induce NF-kappaB activation , Ly294002 ( a PI-3K inhibitor ) , dominant-negative PI-3K , and kinase-dead Akt block IFN-dependent NF-kappaB activation . Moreover , dominant-negative PI-3K blocks IFN-promoted degradation of kappaBox alpha . Ly294002 , a dominant-negative PI-3K construct , and kinase-dead Akt block IFN-promoted cell survival , enhancing apoptotic cell death . Therefore , P40763 , PI-3K , and Akt are components of an IFN signaling pathway that promotes cell survival through NF-kappaB activation . Poly(ADP-ribose) polymerase-1 mediates angiotensin II-induced expression of plasminogen activator inhibitor-1 and fibronectin in rat mesangial cells . OBJECTIVE : To investigate the effects of poly(ADP-ribose) polymerase-1 ( P09874 ) on angiotensin II ( Ang II ) -induced plasminogen activator inhibitor-1 ( P05121 ) and fibronectin ( FN ) in rat mesangial cells ( RMCs ) . METHODS : Followed by serum starvation for 16 h , RMCs were exposed to Ang II for an indicated time to examine the protein expression of P09874 . The cells were treated with or without Ang II for 12-24 h in the presence or absence of an inhibitor of PARP , N-(6-oxo-5,6-dihydrophenanthridin-2-yl)-N,N-dimethylacetamide hydrochloride ( PJ34 ) or small interfering RNA ( siRNA ) duplexes targeting P09874 . The mRNA and protein expressions of P09874 , P05121 and FN were determined by real-time RT-PCR and Western blot , respectively . The activity of P09874 was examined by colorimetric assay . RESULTS : Ang II did not only significantly induce P09874 expression and activity , but also increased P05121 and FN expression in RMCs . All these responses induced by Ang II were significantly inhibited by both the PARP inhibitor PJ34 and downregulating P09874 with the siRNA technique . CONCLUSIONS : Our data suggest that P09874 mediates Ang II-induced P05121 and FN in RMCs and may thus represent a potential therapeutic target in the treatment of glomerular disease . [ Biological function of fusion protein P39905 -PAI2CD ] . To express the fusion protein P39905 -PAI2CD ( urokinase-type plasminogen activator amino terminal fragment-plasminogen activator inhibitor type 2 with the region inter C and D helices deleted ) gene in E.coli and determine the biological characterization of fusion protein P39905 -PAI2CD , the cDNA fragment encoding P39905 -PAI2CD was cloned into the expression vector pLY-4 and transformed into E.coli JF1125 . After temperature induction , the expression amount of P39905 -PAI2CD account for 15 % of total bacterial protein . The result was confirmed by Western blot . P39905 -PAI2CD protein was isolated and purified by washing and solubilization of inclusion body , renaturation and ion exchange chromatography . The final product displayed a single band with a corresponding molecular weight 62 kD in SDS-PAGE . The purity was over 90 % , the protein yield was 50 % and the specific activity was 12 000 IU/mg . The P05121 activity was measured by chromogenic assay . The purified fusion protein inhibited urokinase-type plasminogen activator as measured by milk-agarose plate assay , and bound to human lung cancer cells via uPA receptor ( Q03405 ) , which was confirmed by radio competition experiments . The results indicate that the biological characteristics of P39905 -PAI2CD were very similar to those of the wide type P05120 ( or mutants P05120 , P05121 -2CD ) and to pro-uPA in binding to Q03405 -bearing cells . Interferon-γ promotes vascular remodeling in human microvascular endothelial cells by upregulating endothelin ( ET ) -1 and transforming growth factor ( TGF ) β2 . Systemic sclerosis ( SSc ) is a complex disease characterized by vascular alterations , activation of the immune system and tissue fibrosis . Previous studies have implicated activation of the interferon pathways in the pathogenesis of SSc . The goal of this study was to determine whether interferon type I and/or type II could play a pathogenic role in SSc vasculopathy . Human dermal microvascular endothelial cells ( HDMVECs ) and fibroblasts were obtained from foreskins of healthy newborns . The RT Profiler PCR Array System was utilized to screen for EndoMT genes . Treatment with IFN-α or IFN-γ downregulated Fli1 and P33151 . In contrast , IFN-α and IFN-γ exerted opposite effects on the expression of α-SMA , P29279 , ET-1 , and TGFβ2 , with IFN-α downregulating and IFN-γ upregulating this set of genes . Blockade of TGFβ signaling normalized IFN-γ-mediated changes in Fli1 , P33151 , P29279 , and ET-1 levels , whereas upregulation of α-SMA and TGFβ2 was not affected . DB00559 treatment was more effective than TGFβ blockade in reversing the actions of IFN-γ , including downregulation of α-SMA and TGFβ2 , suggesting that activation of the ET-1 pathway plays a main role in the IFN-γ responses in HDMECs . IFN-γ induced expression of selected genes related to endothelial-to-mesenchymal transition ( EndoMT ) , including Snail1 , P02751 , P05121 , Q15672 , P40763 , P41220 , and components of the WNT pathway . The effect of IFN-γ on EndoMT was mediated via TGFβ2 and ET-1 signaling pathways . This study demonstrates distinct effects of IFN-α and IFN-γ on the biology of vascular endothelial cells . IFN-γ may contribute to abnormal vascular remodeling and fibrogenesis in SSc , partially via induction of EndoMT . Candidate genetic markers and the risk of restenosis after coronary angioplasty . The aim of the present study was to test for possible associations between candidate gene polymorphisms and the risk of restenosis and recurrent restenosis after percutaneous transluminal coronary angioplasty ( PTCA ) without stenting . We followed up 511 PTCA patients , and restenosis and recurrent restenosis were defined according to angiographical criteria . Genotyping of the beta-fibrinogen -455 G/A , glycoprotein ( GP ) IIIa PlA1/PlA2 , plasminogen activator inhibitor-1 ( P05121 ) 4G/5G , factor V Leiden 1691 G/A , tumour necrosis factor alpha ( TNFalpha ) -238 G/A , TNFalpha -308 G/A , interleukin ( IL ) -1alpha -889 C/T , IL-1beta -511 C/T , methylenetetrahydrofolate reductase ( P42898 ) 677 C/T and endothelial nitric oxide synthase ( P29474 ) 4 b/a gene polymorphisms was performed by PCR and restriction-fragment-length-polymorphism-based techniques . One hundred and sixty patients ( 31.3 % ) developed restenosis and in 130 of these patients , of whom 123 were available for analysis , a second PTCA without stenting was performed . Of these patients , 35 ( 28.5 % ) developed recurrent restenosis . None of the investigated genotypes were associated with the risk of restenosis or recurrent restenosis after PTCA . The degree of stenosis before and immediately after PTCA and the severity of the lesion were independent predictors for restenosis after PTCA . In conclusion , there was no association between the beta-fibrinogen -455 G/A , GP IIIa PlA1/A2 , P05121 4G/5G , factor V Leiden 1691 G/A , TNFalpha -238 G/A , TNFalpha -308 G/A , IL-1alpha -889 C/T , the IL-1beta -511 C/T , P42898 677 C/T and P29474 4 b/a gene polymorphisms and the risk of restenosis after PTCA as well as recurrent restenosis after repeated PTCA . Q9Y2D1 polymorphisms influence P39905 function and response to treatment in children with childhood acute lymphoblastic leukemia . DB00023 is a standard and critical component in the therapy of childhood acute lymphoblastic leukemia . DB00174 synthetase ( P08243 ) and the basic region leucine zipper activating transcription factor 5 ( Q9Y2D1 ) and arginosuccinate synthase 1 ( P00966 ) have been shown to mediate the antileukemic effect of asparaginase and to display variable expression between leukemia cells that are resistant and sensitive to treatment . Fourteen polymorphisms in the regulatory and coding regions of these genes were investigated for an association with acute lymphoblastic leukemia outcome . Lower event-free survival ( O43281 ) was associated with Q9Y2D1 T1562C , tandem-repeat P08243 polymorphism , derived haplotype , and P00966 G1343T and G34T substitutions ( P ≤ .03 ) . Associations were limited to patients who received Escherichia coli asparaginase . Variations that sustained correction for multiple testing ( Q9Y2D1 T1562C , P = .005 ; P08243 tandem-repeat and related haplotype , P ≤ .01 ) were subsequently analyzed in the replication cohort . The E coli-dependent association of the Q9Y2D1 T1562 allele with reduced O43281 was confirmed ( P = .01 ) . A gene-reporter assay showed that the haplotype tagged by T1562 had higher promoter activity ( P ≤ .01 ) . The remaining regulatory polymorphisms also appeared to affect Q9Y2D1 function ; 2 additional high-activity haplotypes were identified ( P ≤ .02 ) and were further corroborated by quantitative mRNA analysis in lymphoblastoid cell lines . The Q9Y2D1 -regulated increase in P08243 expression in response to more efficacious E coli-induced asparagine depletion may explain our observed results . Endothelin receptor antagonists as disease modifiers in systemic sclerosis . Systemic sclerosis ( SSc ) is a multisystem connective tissue disease of unknown etiology that is characterized by inflammation , vascular dysfunction and fibrosis of the skin and visceral organs . SSc is clinically diverse both in terms of the burden of skin and organ involvement and the rate of progression of the disease . Recent studies indicate that the endothelin system , especially ET-1 and the P25101 and ETB receptors may play a key role in the pathogenesis of SSc . A new class of drugs , endothelin receptor antagonists has been introduced for treatment of patients with pulmonary arterial hypertension ( PAH ) . DB00559 , a dual endothelin receptor antagonist as well as DB06268 and DB06403 , selective blockers of the P25101 receptor have proven effective in SSc-PAH . This effect may be mediated through both a vasodilatory and antifibrotic effect , thus making these agents attractive as potential disease modifying agents for SSc . Anti-angiogenic effects of interleukin-12 delivered by a novel hyperthermia induced gene construct . PURPOSE : Interleukin-12 ( IL-12 ) is a pro-inflammatory cytokine possessing anti-cancer and anti-angiogenic properties . This study quantitatively assessed the anti-angiogenic effect of IL-12 delivered using an adenoviral vector with murine IL-12 placed under control of a heat shock promoter . This approach limits systemic toxicity by restricting IL-12 delivery locally to the tumour . The kinetics of the downstream cytokines interferon-gamma ( P01579 ) and interferon inducible protein-10 ( P02778 ) and other molecules affecting angiogenesis , vascular endothelial growth factor ( P15692 ) and plasminogen activator inhibitor-1 ( P05121 ) were also studied . MATERIALS AND METHODS : 4T1 tumours were grown in Balb/C mice and the AdhspmIL-12 construct was injected intra-tumourally . The tumours were heated after 24 h using a water bath . At various time points post-heating the tumours were collected and quantitatively assessed for cytokine production and vascularity . RESULTS : A significant reduction was seen in the tumour vasculature of the treated group vs. the control group mice . Systemic effects of IL-12 were limited to generalized immunostimulation . No hepatoxicity was noted . CONCLUSIONS : This study suggests that IL-12 can be effectively delivered using a gene-based approach with a heat shock promoter . This results in quantitatively measurable anti-angiogenesis and general immunostimulation . The complex inter-play of other pro- and anti-angiogenic factors ( P01579 , P02778 , P15692 and P05121 ) was also studied . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . [ Endothelial progenitor cells related gene expression changes before and early after revascularization in patients with acute myocardial infarction ] . OBJECTIVE : The purpose of this study was to observe the endothelial progenitor cells ( EPCs ) related gene expression changes before and early after revascularization in patients with acute myocardial infarction . METHODS : Peripheral blood samples were taken from patients with acute anterior myocardial infarction 6 hours and 7 days after P05154 and stenting . Mononuclear cells ( MNCs ) were isolated by Ficoll-density centrifugation and cultured in M-199 medium . After 14 days culture , attaching cells incorporated DiI-acetylated low-density lipoprotein ( EPCs ) were collected and RNA was isolated by Trizol for microarray analysis on 24 genes associated with permissibility/vessel tone ( angiotensin system : P12821 , AGTR-1 , AGTR-2 ; NO system : P29474 ; prostacyclin system : P35354 ; endothelin system : ET-1 , P25101 , ETB ; superoxide anions system : SOD-1 ) , angiogenesis ( adhesion molecule : P33151 ; growth factors and receptors : P17948 , P35968 , P15692 ) and endothelial cell activation ( adhesion molecules expression : P05362 , P13598 , P32942 , P16284 , E-Selectin , L-Selection , P19320 ; change phenotype from antithrombotic to prothrombotic : tPA , uPA , P05121 , P04275 ) . P35968 , P16284 and P33151 positive cells were identified by flow cytometry . RESULT : Eight gene expressions ( AGTR-1 , AGTR-2 , P35354 , P29474 , ET-1 , P25101 , P15692 ) were significantly downregulated 7 days post P05154 compared to pre- P05154 ( P < 0.05 ) . Flow cytometry results showed that P35968 positive cells were also significantly reduced post P05154 than that of before P05154 ( P < 0.05 ) . CONCLUSION : P05154 down-regulated endothelial progenitor cells related gene expressions in patients with acute myocardial infarction . Potent and selective inhibition of human nitric oxide synthases . Inhibition by non-amino acid isothioureas . DB02539 was a potent competitive inhibitor of human nitric oxide synthase ( NOS ) , with Ki values of 17 , 36 , and 29 nM for the inducible ( i ) , endothelial ( e ) , and neuronal ( n ) isozymes , respectively . Unlike some potent inhibitors of NOS , no time dependence was observed . DB02539 was not a detectable substrate for P29474 . DB02539 was also a potent inhibitor of mouse P35228 ( Ki value of 5.2 nM ) , and its binding perturbed the spectrum of P35228 consistent with its altering the environment of the bound heme . The optimum binding of S-ethyl- and S-isopropylisothiourea relative to 70 other analogs suggested that these alkyl substitutions fit into a small hydrophobic pocket . Most isothioureas were 2-6-fold selective for the human P35228 ( Ki for P35228 versus Ki for P29474 ) , with one being 19-fold selective . The cyclized mimics of S-ethylisothiourea , 2-NH2-thiazoline , and 2-NH2-thiazole , were also competitive inhibitors of human NOS . A third structural class of inhibitors , bisisothioureas , were , in general , the most selective in their inhibition of human P35228 . S,S'-(1,3-Phenylenebis(1,2-ethanediyl))bisisothiourea was 190-fold selective ( Ki value of 0.047 microM against P35228 versus 9.0 microM against P29474 ) . These results demonstrate that potent and selective inhibition of human NOS isozymes is achievable . Amelioration of nephropathy with apoA-1 mimetic peptide in apoE-deficient mice . BACKGROUND : There is mounting evidence that dyslipidaemia may contribute to development and progression of renal disease . For instance , hyperlipidaemia in apolipoprotein E-deficient ( apoE(-/-) ) mice is associated with glomerular inflammation , mesangial expansion and foam cell formation . ApoA-1 mimetic peptides are potent antioxidant and anti-inflammatory compounds which are highly effective in ameliorating atherosclerosis and inflammation in experimental animals . Given the central role of oxidative stress and inflammation in progression of renal disease , we hypothesized that apoA-1 mimetic peptide , D-4F , may attenuate renal lesions in apoE(-/-) mice . METHODS : Twenty-five-month-old female apoE(-/-) mice were treated with D-4F ( 300 µg/mL in drinking water ) or placebo for 6 weeks . Kidneys were harvested and examined for histological and biochemical characteristics . RESULTS : Compared with the control mice , apoE(-/-) mice showed significant proteinuria , tubulo-interstitial inflammation , mesangial expansion , foam cell formation and up-regulation of oxidative [ NAD(P)H oxidase subunits ] and inflammatory [ NF-κB , P13500 , P05121 and P35354 ] pathways . D-4F administration lowered proteinuria , improved renal histology and reversed up-regulation of inflammatory and oxidative pathways with only minimal changes in plasma lipid levels . CONCLUSIONS : The apoE(-/-) mice develop proteinuria and glomerular and tubulo-interstitial injury which are associated with up-regulation of oxidative and inflammatory mediators in the kidney and are ameliorated by the administration of apoA-1 mimetic peptide . These observations point to the role of oxidative stress and inflammation in the pathogenesis of renal disease in hyperlipidaemic animals and perhaps humans . Serotonin induces the expression of tissue factor and plasminogen activator inhibitor-1 in cultured rat aortic endothelial cells . Serotonin ( 5-hydroxytryptamine , or 5-HT ) , released from activated platelets , not only accelerates aggregation of platelets but also is known to promote mitosis , migration , and contraction of vascular smooth muscle cells ( VSMCs ) . These effects are considered to contribute to thrombus formation and atherosclerosis . The aim of this study was to investigate the effects of 5-HT on the expressions of coagulative and fibrinolytic factors in rat aortic endothelial cells . Endothelial cells were stimulated with various concentrations of 5-HT ( 0.1 approximately 10 microM ) , and the expressions of tissue factor ( TF ) , tissue factor pathway inhibitor ( P10646 ) , plasminogen activator inhibitor-1 ( P05121 ) , and tissue-type plasminogen activator ( TPA ) messenger RNAs ( mRNAs ) were evaluated by Northern blot analysis . The activities of TF and P05121 were also measured . TF and P05121 mRNA were increased significantly in a concentration- and time-dependent manner . However , P10646 and TPA mRNA expression did not change . The inductions of TF and P05121 mRNAs were inhibited by a 5-HT1/5-HT2 receptor antagonist ( methiothepin ) and a selective 5- Q13049 receptor antagonist ( D6RGH6 -9042 ) . These results indicate that 5-HT increases procoagulant activity and reduces fibrinolytic activities of endothelial cells through the 5- Q13049 receptor . It was concluded that the modulation of procoagulant and hypofibrinolytic activities of endothelial cells by 5-HT synergistically promotes thrombus formation at the site of vessel injury with the platelet aggregation , VSMC contraction , and VSMC proliferation . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . Effect of the R(-) and S(+) isomers of MDA and DB01454 on phosphatidyl inositol turnover in cultured cells expressing 5- Q13049 or P28335 receptors . The effect of the R(-) and S(+) isomers of 3,4-methylenedioxyamphetamine ( MDA ) and its N-methyl analog 3,4-methylenedioxymethamphetamine ( DB01454 ) on [3H]inositol monophosphate accumulation was studied in cells expressing either 5- Q13049 or P28335 receptors . The isomers of MDA produced a concentration dependent increase in phosphatidyl inositol ( PI ) hydrolysis at the 5- Q13049 receptors , with the R(-) isomer of MDA being more potent than the S(+) at the 5- Q13049 receptor . The R(-) and S(+) isomers of DB01454 were significantly less efficacious at the 5- Q13049 receptor as compared to MDA ; S(+) DB01454 had no effect . At the P28335 receptor , both R(-) and S(+)MDA were equipotent at stimulating PI hydrolysis , with the S(+) isomer of DB01454 being more efficacious at the P28335 receptor compared with the R(-) isomer . In all cases at both the 5- Q13049 and P28335 receptors , the affinities of the isomers of DB01454 and MDA were at least 2-3 orders of magnitude less than 5-HT . Despite the weak effect of these compounds at the 5- Q13049 and P28335 receptors , these substituted amphetamines do possess intrinsic activity which may contribute to their neurotoxic effects when administered at high doses . CCK1-receptor stimulation protects against gut mediator-induced lung damage during endotoxemia . BACKGROUND/AIMS : Cholecystokinin 1-receptor ( P32238 ) activation by long chain fatty acid ( LCFA ) absorption stimulates vago-vagal reflex pathways in the brain stem . The present study determines whether this reflex also activates the cholinergic anti-inflammatory pathway , a pathway known to modulate cytokine release during endotoxemia . METHODS : Mesenteric lymph was obtained from wild type ( WT ) and P32238 knockout ( P32238 (-/-) ) mice intraperitoneally challenged with Lipopolysaccharid ( LPS ) ( endotoxemic lymph , EL ) and intestinally infused with vehicle or LCFA-enriched solution . The lymph was analyzed for TNFα , P05231 and P22301 concentration and administered to healthy recipient mice via jugular infusion . Alveolar wall thickness , myeloperoxidase ( P05164 ) and TUNEL positive cells were determined in lung tissue of recipient mice . RESULTS : LCFA infusion in WT mice reduced TNFα concentration in EL by 49 % compared to vehicle infusion , but had no effect in P32238 (-/-) mice . EL significantly increased the alveolar wall thickness , the number of P05164 -positive and TUNEL-positive cells compared to control lymph administration . LCFA infusion in WT , but not in CCK1R(-/-) mice , significantly reduced these pathological effects of EL . CONCLUSION : During endotoxemia enteral LCFA absorption reduces TNFα release into mesenteric lymph and attenuates histomorphologic parameters of lung dysfunction . Failure to elicit this effect in CCK1R(-/-) mice demonstrates that anti-inflammatory properties of LCFAs are mediated through CCK1-Rs . Evaluation of pharmacological profile of meloxicam as an anti-inflammatory agent , with particular reference to its relative selectivity for cyclooxygenase-2 over cyclooxygenase-1 . We studied the anti-inflammatory activity of meloxicam on rat carrageenin-induced pleurisy and its toxicity for rat gastric mucosa , relative to its in vitro inhibitory potency against partially purified cyclooxygenase ( P36551 ) -1 and P35354 preparations in order to clarify the pharmacological profile of the compound as an anti-inflammatory agent . In rat carrageenin-induced pleurisy , the plasma exudation rate peaked at 5 h , at which time P35354 was detectable in cells from the pleural exudate . Meloxicam and piroxicam ( 1 and 3 mg/kg ) and NS-398 ( 3 mg/kg ) showed almost equal anti-inflammatory potency against 5-hour pleurisy . A single oral administration of the compounds caused a dose-dependent increase in the number of rats with gastric mucosal erosion . The ED50 value for meloxicam ( 5.92 mg/kg ) was significantly higher than that for piroxicam ( 1.76 mg/kg ) , indicating that meloxicam is safer . DB00328 showed intermediate safety ( 2.59 mg/kg ) . In in vitro experiments , indometacin inhibited P23219 about 1.7 times more potently than P35354 . NS-398 inhibited P35354 with an IC50 of 0.32 microM , but never affected P23219 activity , even at 100 microM . In the same assay system , meloxicam inhibited P35354 about 12 times more selectively than P23219 . Piroxicam , however , inhibited both isoforms almost equally . These results indicate that meloxicam is a potent anti-inflammatory agent with low gastric toxicity . One reason for its in vivo pharmacological profile may be related to its relative selectivity for P35354 over P23219 . Thus , meloxicam may belong to a group of P35354 selective anti-inflammatory agents with a better safety profile than conventional P23219 and P35354 nonselective anti-inflammatory agents . Cholecystokinin-related peptides , after systemic or central administration , prevent carbon monoxide-induced amnesia in mice . The neuroprotective actions of cholecystokinin ( CCK ) peptides were investigated in a mouse hypoxia model , in which the animals were successively exposed to CO gas . Working memory impairment 5 days after CO exposure was examined by using a Y-maze test ; delayed amnesia was examined 7 days after CO exposure , by using a step-down type passive avoidance test . DB00403 ( 1-100 micrograms/kg , given s.c. 30 min before CO exposure ) significantly prevented the CO-induced impairment of performance in both tests , the improvement being correlated with the severity of hypoxia . This severity was increased by maintaining the body temperature at 38 degrees C . DB00403 was less effective when injected immediately after a single CO exposure . The order of potency of the CCK-peptides administered systemically was : ceruletide > CCK-8S > CCK-8NS >> Q13308 . DB00403 ( 0.03-0.3 micrograms/mouse ) and CCK-8S ( 0.03-1 microgram/mouse ) prevented CO-induced amnesia after i.c.v. administration . Under all experimental conditions , dizocilpine [ MK-801 , (+)-5-methyl-10,11-dihydro-5H-dibenzo(a,d)cyclohepten-5,10-imine maleate , 500 micrograms/kg s.c. or 10 micrograms/mouse i.c.v. ] prevented completely the CO-induced amnesia . The protective effects of systemic ceruletide were blocked , partially but significantly , by the preadministration of L-364,718 ( 3S-(-)-N- [ 2,3-dihydro-1-methyl-2-oxo-S-phenyl-1H-1,4- benzodiazepine-3-yl ] -1H-indole-2-carboxamide , 1-10 mg/kg i.p. ) , a selective P32238 antagonist . L-365,260 ( [ 3R-(+)-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepine-3-yl ] -N ' - [3-methyl-phenyl]urea ) , a CCK-B antagonist , also decreased ceruletide-induced protection. ( ABSTRACT TRUNCATED AT 250 WORDS ) Detrimental role of endogenous nitric oxide in host defence against Sporothrix schenckii . We earlier demonstrated that nitric oxide ( NO ) is a fungicidal molecule against Sporothrix schenckii in vitro . In the present study we used mice deficient in inducible nitric oxide synthase ( P35228 -/- ) and C57BL/6 wild-type ( WT ) mice treated with Nomega-nitro-arginine ( Nitro- DB00125 -treated mice ) , an NOS inhibitor , both defective in the production of reactive nitrogen intermediates , to investigate the role of endogenous NO during systemic sporotrichosis . When inoculated with yeast cells of S. schenckii , WT mice presented T-cell suppression and high tissue fungal dissemination , succumbing to infection . Furthermore , susceptibility of mice seems to be related to apoptosis and high interleukin-10 and tumour necrosis factor-alpha production by spleen cells . In addition , fungicidal activity and NO production by interferon-gamma ( P01579 ) and lipopolysaccharide-activated macrophages from WT mice were abolished after fungal infection . Strikingly , P35228 -/- and Nitro- DB00125 -treated mice presented fungal resistance , controlling fungal load in tissues and restoring T-cell activity , as well as producing high amounts of P01579 Interestingly , macrophages from these groups of mice presented fungicidal activity after in vitro stimulation with higher doses of P01579 . Herein , these results suggest that although NO was an essential mediator to the in vitro killing of S. schenckii by macrophages , the activation of NO system in vivo contributes to the immunosuppression and cytokine balance during early phases of infection with S. schenckii . Leishmania mexicana amazonensis : ADP-ribosyltransferase antagonists specifically inhibit amastigote to promastigote differentiation . Leishmania mexicana amazonensis amastigotes were induced to differentiate by incubation at 27 C . Morphological transformation was studied both in untreated cultures and in cultures where DNA synthesis , and consequently the final stage in the production of promastigotes , was inhibited by hydroxyurea . DB03073 and other antagonists of ADP-ribosyltransferase ( P09874 ) specifically inhibited differentiation at a very early stage in both experimental systems . Cell proliferation ( in the absence of hydroxyurea ) was not inhibited by P09874 antagonists -- indeed greater multiplication of undifferentiated parasites was observed in the presence of these compounds . This indicated that the parasites were being diverted from differentiation to proliferation . Preincubation of the amastigotes with the P09874 antagonists was required to produce this effect , providing further evidence that ADP-ribosylation of proteins is required for the initiation of differentiation in Leishmania .

MH_train_1305

MH_train_1305

MH_train_1305

interacts_with DB00904?

multiple_choice

Local immunotherapy of glioma patients with a combination of 2 bispecific antibody fragments and resting autologous lymphocytes : evidence for in situ t-cell activation and therapeutic efficacy . After adoptive transfer of pre-activated lymphocytes into the operation cavity of glioma patients , tumor regression and improved survival have been reported in some patients . Results were most impressive when bispecific antibodies with tumor x CD3 specificity were also applied . In this study , we attempted to avoid time-consuming pre-activation procedures for adoptively transferred cells by using a combination of bispecific antibodies directed to the P01133 receptor ( P00533 ) on tumor cells and to CD3 and P10747 on T cells . Eleven patients with high-grade malignant glioma received 3 injections of 2 bispecific antibody fragments ( P00533 x CD3 and P00533 x P10747 ) together with freshly isolated autologous lymphocytes via an Ommaya reservoir . Intracavitary fluid aspirated during immunotherapy was examined for markers of T-cell activation . Increased levels of soluble P60568 receptor and P01375 were detected in the intracavitary fluid of all patients tested . Two of the 11 treated patients experienced a beneficial response to therapy as defined by a transient contrast enhancement in subsequent Q9BWK5 scans and prolonged survival . Side effects were transient and consisted of fever , nausea , headache and aggravation of pre-existing neurologic deficits . These adverse effects were most likely due to the antibody construct containing anti-CD3 specificity . Two patients developed cerebral edema and required steroid treatment . The role of the polyol pathway in acute kidney injury caused by hindlimb ischaemia in mice . The polyol pathway , a collateral glycolytic process , previously considered to be active in high glucose milieu , has recently been proposed to play a crucial role in ischaemia/reperfusion tissue injury . In this study , we explored the role of the polyol pathway in acute kidney injury ( AKI ) , a life-threatening condition , caused by hindlimb ischaemia , and determined if inhibition of the polyol pathway by aldose reductase ( AR ) inhibitor is beneficial for this serious disorder . Mice 8 weeks of age rendered hindlimb ischaemic for 3 h by the clipping of major supporting arteries revealed marked muscle necrosis with accumulation of sorbitol and fructose in ischaemic muscles . Serum concentrations of blood urea nitrogen ( BUN ) , creatinine phosphokinase ( CPK ) , creatinine , tumour necrosis factor ( P01375 ) -alpha as well as interleukin ( IL ) -6 were all elevated in these mice . Treatment with AR inhibitor ( Q9Y4X5 ) effectively suppressed muscle necrosis and accompanying inflammatory reactions and prevented renal failure . Similar to Q9Y4X5 -treated mice , AR-deficient mice were protected from severe ischaemic limb injury and renal failure , showing only modest muscle necrosis and significant suppression of serum markers of renal failure and inflammation . Thus , these findings suggest that the polyol pathway is implicated in AKI caused by ischaemic limb injury and that AR may be a potential therapeutic target for this condition . Targeted therapies for adrenocortical carcinoma : IGF and beyond . Standard chemotherapy for adrenocortical cancer currently is under evaluation in the context of the recently completed FIRM-ACT evaluating the combination of mitotane with either streptozocin or etoposide , cisplatin , and doxorubicin . New agents are eagerly sought by the ACC community that hopes to make progress against this deadly disease . Investigators have begun to dissect the molecular and genomic context of ACC with a goal of identifying potential novel therapeutic agents . One gene consistently overexpressed in ACC is insulin growth factor type 2 . Targeting its receptor P08069 has shown encouraging results in ACC cell lines and against murine xenografts . As a result , clinical trials to evaluate agents targeting the P08069 have been done including mitotane and DB05759 ( a monoclonal antibody ) and the GALACCTIC trial that has just completed accrual to evaluate OSI-906 , a small molecule P08069 antagonist . On the horizon are other agents targeting other tyrosine kinases , including P01133 and FGF , and novel strategies such as individualized tumor analysis to select treatment . Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . Anticytokine treatment prevents the increase in the activity of DB00171 -ubiquitin- and Ca(2+)-dependent proteolytic systems in the muscle of tumour-bearing rats . The ascites hepatoma Yoshida AH-130 induces loss of body weight and tissue waste . Tumour necrosis factor alpha ( P01375 ) plays a pivotal role in the pathogenesis of muscle wasting in this model system , but other cytokines , such as interleukin-6 , may be involved . In order to verify whether a combined anticytokine treatment may synergistically counteract muscle protein degradation , tumour bearing rats were treated with pentoxyfilline ( PTX , an inhibitor of P01375 synthesis ) , or with suramin ( Q09428 , an antiprotozoal drug blocking the peripheral action of several cytokines including P05231 and P01375 ) , or both the drugs , and the effects on muscle proteolytic systems were assessed . Muscle protein loss in the AH-130-bearing rats was associated with increased activity of both the DB00171 -ubiquitin- and the calpain- dependent proteolytic pathways ( 246 % and 230 % of controls , respectively ) . Both PTX and Q09428 , either alone or in combination , prevented the depletion of muscle mass and significantly reduced the activity of muscle proteolytic systems . In particular , treatment with Q09428 , either alone or with PTX , induced a decrease in enzymatic activities to values similar to those of controls . The results obtained in the present paper demonstrate that : ( i ) muscle depletion in this model is indeed associated with increased proteasome- and calpain-dependent proteolysis , as previously suggested by increased mRNA expression of molecules pertaining to both pathways ; ( ii ) anticytokine treatments effectively reduce muscle protein loss by down-regulating the activity of at least two major proteolitic systems ; ( iii ) Q09428 is more effective than PTX in reducing the activity of proteolytic systems , possibly because of its multiple anticytokine action . Determination of the class and isoform selectivity of small-molecule histone deacetylase inhibitors . The human HDAC ( histone deacetylase ) family , a well-validated anticancer target , plays a key role in the control of gene expression through regulation of transcription . While HDACs can be subdivided into three main classes , the class I , class II and class III HDACs ( sirtuins ) , it is presently unclear whether inhibiting multiple HDACs using pan-HDAC inhibitors , or targeting specific isoforms that show aberrant levels in tumours , will prove more effective as an anticancer strategy in the clinic . To address the above issues , we have tested a number of clinically relevant HDACis ( HDAC inhibitors ) against a panel of rhHDAC ( recombinant human HDAC ) isoforms . Eight rhHDACs were expressed using a baculoviral system , and a Fluor de Lystrade mark ( Biomol International ) HDAC assay was optimized for each purified isoform . The potency and selectivity of ten HDACs on class I isoforms ( rhHDAC1 , rhHDAC2 , rhHDAC3 and rhHDAC8 ) and class II HDAC isoforms ( rhHDAC4 , rhHDAC6 , rhHDAC7 and rhHDAC9 ) was determined . MS-275 was Q13547 -selective , DB05651 was Q13547 - and Q92769 -selective , apicidin was Q92769 - and O15379 -selective and valproic acid was a specific inhibitor of class I HDACs . The hydroxamic acid-derived compounds ( trichostatin A , DB00238 -LAQ824 , DB06603 , ITF2357 , vorinostat and belinostat ) were potent pan-HDAC inhibitors . The growth-inhibitory effect of the HDACis on HeLa cells showed that both pan-HDAC and class-I-specific inhibitors inhibited cell growth . The results also showed that both pan-HDAC and class-I-specific inhibitor treatment resulted in increased acetylation of histones , but only pan-HDAC inhibitor treatment resulted in increased tubulin acetylation , which is in agreement with their activity towards the Q9UBN7 isoform . Attenuation of the progression of adjuvant-induced arthritis by 3-aminobenzamide treatment . Rheumatoid arthritis ( RA ) is a disease that is still insufficiently controlled by current treatments . Poly(ADP-ribose) polymerase ( PARP ) inhibitors ameliorate immune-mediated diseases in several experimental models , including RA , colitis , experimental autoimmune encephalomyelitis and allergy . Together these findings showed that ADP-ribosylating enzymes , in particular P09874 , play a pivotal role in the regulation of immune responses and may represent a noble target for new therapeutic approaches in immune-mediated diseases . The effect of 3-aminobenzamide ( 3-AB ) , an inhibitor of poly(ADP-ribose) synthetase activity , was evaluated in a mouse model of adjuvant-induced arthritis ( AIA ) on pro-inflammatory cytokines , adhesion molecules , inflammatory mediators and chemokine production/expression in serum and knee joint . Histopathological examination was also done on joint section . Our data demonstrates that 3-AB , 10mg/kg , intraperitoneally ( i.p. ) significantly reduces pro-inflammatory cytokine ( Q16552 , P01375 -α and P60568 ) and chemokine ( P13500 and MIP-2 ) production/expression , accompanied by amelioration of the disease as indicated by reduced paw swelling and arthritic scores and was associated with a significant reduction of P19320 and P05362 expression in the knee joint . Moreover , the expression of inflammatory mediators ( P35228 , P35354 , P08253 , P14780 ) and joint histological inflammatory damage was also markedly decreased . The results of this study suggest that P09874 inhibitor may play a role in the inflammatory arthritic process after administration of 3-AB may be a beneficial therapeutic approach . In vitro cell death of activated lymphocytes in Omenn 's syndrome . Omenn 's syndrome ( OS ) is characterized by erythrodermia , hepatosplenomegaly , lymphadenopathy , hypereosinophilia and elevated IgE levels associated with increased susceptibility to severe infections . Peripheral blood T cells , though usually present in normal number , show an activated phenotype ( including an increased expression of CD95/Fas ) , a Th2 pattern of cytokine secretion and defective proliferative response to mitogens . In this report , we demonstrate that T cells from patients with OS undergo an excessive cell death in vitro resulting from two mechanisms . First , a substantial number of peripheral blood lymphocytes from OS patients die in unstimulated cultures ( p = 0.009 vs. healthy controls ) . This spontaneous apoptosis is associated with reduced expression of bcl-2 gene product ( p < 0.05 ) and can be prevented by addition of interleukin ( IL ) -2 ( which also prevents down-modulation of bcl-2 ) , while is independent from CD95 signaling . Second , lymphocytes from OS patients are highly susceptible to activation-induced cell death ( AICD ) induced with mitogens . This mechanism is largely independent from P60568 , while it can be significantly inhibited blocking CD95 with an IgG2b monoclonal antibody ( mAb ) . The dependence of AICD from signals transduced via CD95 was confirmed showing that cross-linking CD95 with an IgM mAb induces a higher cell death in purified P01730 + CD45R0+ cells from OS patients than in controls ( comparable for CD95 expression ) . Both mechanisms of cell death observed in this study result from lymphocyte hyperactivation occurring in vivo in these patients and may contribute to functional T cell defects of OS . Structure and function of eritadenine and its 3-deaza analogues : potent inhibitors of S-adenosylhomocysteine hydrolase and hypocholesterolemic agents . d- DB03769 ( DEA ) is a potent inhibitor of S-adenosyl-l-homocysteine hydrolase ( P23526 ) and has hypocholesterolemic activity . We have hypothesized that 3-deaza-DEA ( P01024 -DEA ) and its analogues retain high level of P23526 inhibitory activity and have resistance to deamination and glycosidic bond hydrolysis in vivo . Such P01024 -DEA analogues would have much higher hypocholesterolemic activity . P01024 -DEA , and its methyl ester ( P01024 -OMeDEA ) and its methyl amido ( P01024 -NMeDEA ) were synthesized to examine their P23526 inhibitory and hypocholesterolemic activities . A crystal structure of P23526 containing P01024 -DEA was determined and confirmed that DEA and P01024 -DEA bound to the same site of P23526 with the same binding mode . The P23526 inhibitory activities of P01024 -DEA ( K(I)=1.5 microM ) and P01024 -OMeDEA ( K(I)=1.5 microM ) are significantly lower than that of DEA ( K(I)=30 nM ) , while rats fed by P01024 -DEA and P01024 -OMeDEA decrease the total plasma cholesterol and phospholipids by 36-40 % and 23 % , respectively , which is similar to the level of reductions ( 42 % and 27 % ) by DEA . P01024 -NMeDEA lost most of the P23526 inhibitory activity ( K(I)=30 microM ) and dietary P01024 -NMeDEA does not decrease cholesterol and phospholipid in plasma but decreases the triacylglycerol level by 16 % . DEA and P01024 -DEA analogues are neither substrates nor inhibitors of adenosine deaminase . MR properties of excised neural tissue following experimentally induced inflammation . Changes in the MR parameters of inflamed neural tissue were measured in vitro . P01375 -alpha ( P01375 ) was injected into rat sciatic nerves to induce inflammation with negligible axonal loss and demyelination . The MR parameters , such as T1/ P24752 relaxation and magnetization transfer ( MT ) , were measured 2 days after P01375 injection and were found to be substantially different from those of normal nerves . The average T1/ P24752 relaxation times increased , whereas the MT ratio ( Q99707 ) and the quantitative MT parameter M0B ( which describes the semisolid pool of protons ) decreased . The MR parameters correlated very well with the extracellular volume fraction ( EM ) of neural tissue evaluated by quantitative histopathology . The multicomponent P24752 relaxation was shown to provide the best quantitative assessment of changes in neural tissue microstructure , and allowed us to distinguish between the processes of inflammation and demyelination . In comparison , the MT measurements were less successful due to competing contributions of demyelination and pH-sensitive changes in the MT effect . Functional methionine synthase deficiency ( cblE and cblG ) : clinical and biochemical heterogeneity . Functional methionine synthase deficiency is generally characterized by homocystinuria and hypomethioninemia in the absence of methylmalonic aciduria . Patients are divided into two classes , cblE and cblG , on the basis of complementation analysis . Presentation has usually been in the first 2 years of life , but one patient came to medical attention at age 21 years with symptoms initially diagnosed as multiple sclerosis . Common findings among 11 patients ( 4 with cblE and 7 with cblG ) have included megaloblastic anemia ( all patients ) and various neurological deficits including developmental retardation ( 10 patients ) , cerebral atrophy ( 8 patients ) , hypotonia ( 7 patients ) , EEG abnormalities ( 6 patients ) , and nystagmus ( 5 patients ) . Hypertonia , seizures , blindness , and ataxia were less frequent . All patients have responded to therapy with cobalamin with resolution of anemia and biochemical abnormalities ; neurological deficits resolved more slowly and in some cases incompletely . DB00200 has been more effective than cyanocobalamin . Fibroblasts from patients with cblE ( 5 patients ) and cblG ( 6 patients ) all showed decreased intracellular levels of methylcobalamin ( DB03614 ) and decreased incorporation of label from 5-methyltetrahydrofolate into macromolecules , suggesting decreased activity of the DB03614 -dependent enzyme methionine synthase . Q99707 specific activity in extracts of all cblE fibroblasts was normal or near-normal under standard reducing conditions ; synthase specific activity in extracts of 5 cblG patients was low but was high in a 6th patient measured in another laboratory . Thus , there is heterogeneity among patients with functional methionine synthase deficiency both in clinical presentation and in the results of biochemical studies of cultured cells . [ The importance of determination of interleukin-10 in the blood of patients with systemic lupus erythematosus ] . BACKGROUND : Hitherto , not very numerous investigations provided so far only few often controversial findings on the importance of interleukin-10 ( P22301 ) in systemic lupus erythematosus ( SLE ) . The objective of the present investigation was to assess whether there exist practically applicable relations between the serum level of P22301 , clinical and laboratory indicators of activity of the disease and serum levels of selected cytokines or their soluble receptors . METHODS AND RESULTS : The authors analyzed a group of 23 patients with SLE ( 23 women and 1 man , median age 37 years ) . P22301 and other cytokines were examined by the ELISA method , the clinical activity of the disease was evaluated by the ECLAM system ( European Consensus Lupus Activity Measurement ) . Elevated P22301 values ( > 5 pg/ml ) were assessed in 10 ( 43 % ) patients . Correlation analysis ( Pearson 's test , p < 0.05 ) revealed statistically significant relations between P22301 levels and the activity of the disease , values of antibody levels against dsDNA and levels of the soluble receptor P60568 ( sIL-2R ) in serum . Conversely , no relationship was revealed between values of P22301 and values of P01024 and C4 complement components , IL-1 , P60568 , P05231 , sIL-6R , P01375 , sTNFR-alpha and P27352 -gamma . CONCLUSIONS : Elevated P22301 serum levels in patients with SLE did not have , with the exception of the index of clinical activity of the disease , antibodies against dsDNA and sIL-2R any statistically significant relations to laboratory indicators of disease activity and levels of selected cytokines and their soluble receptors . Q14703 lyase in thymic perivascular spaces promotes egress of mature thymocytes via up-regulation of P21453 . DB03203 1-phosphate ( Q14703 ) and P21453 ( P21453 ) play an important role in the egress of mature P01730 or CD8 single-positive ( SP ) thymocytes from the thymus . DB08868 hydrochloride ( FTY720 ) , an P21453 functional antagonist , induced significant accumulation of CD62L(high) Q07108 (low) mature SP thymocytes in the thymic medulla . Immunohistochemical staining using anti- P21453 antibody revealed that P21453 is predominantly expressed on thymocytes in the thymic medulla and is strongly down-regulated even at 3h after FTY720 administration . 2-Acetyl-4-tetrahydroxybutylimidazole ( THI ) , an Q14703 lyase inhibitor , also induced accumulation of mature SP thymocytes in the thymic medulla with an enlargement of the perivascular spaces ( P15151 ) . At 6h after THI administration , P21453 -expressing thymocytes reduced partially as if to form clusters and hardly existed in the proximity of CD31-expressing blood vessels in the thymic medulla , suggesting Q14703 lyase expression in the cells constructing thymic medullary P15151 . To determine the cells expressing Q14703 lyase in the thymus , we newly established a mAb ( YK19-2 ) specific for mouse Q14703 lyase . Immunohistochemical staining with YK19-2 revealed that Q14703 lyase is predominantly expressed in non-lymphoid thymic stromal cells in the thymic medulla . In the thymic medullary P15151 , Q14703 lyase was expressed in ER-TR7-positive cells ( reticular fibroblasts and pericytes ) and CD31-positive vascular endothelial cells . Our findings suggest that Q14703 lyase expressed in the thymic medullary P15151 keeps the tissue Q14703 concentration low around the vessels and promotes thymic egress via up-regulation of P21453 . Compound heterozygous mutations in the Q09428 ( ABCC 8 ) subunit of pancreatic K( DB00171 ) channels cause neonatal diabetes by perturbing the coupling between Kir6.2 and Q09428 subunits . KATP channels regulate insulin secretion by coupling β-cell metabolism to membrane excitability . These channels are comprised of a pore-forming Kir6.2 tetramer which is enveloped by four regulatory Q09428 subunits . DB00171 acts on Kir6.2 to stabilize the channel closed state while ADP ( coordinated with Mg(2+) ) activates channels via the Q09428 domains . Aberrations in nucleotide-binding or in coupling binding to gating can lead to hyperinsulinism or diabetes . Here , we report a case of diabetes in a 7-mo old child with compound heterozygous mutations in Q09428 ( Q09428 [A30V] and Q09428 [G296R] ) . In unison , these mutations lead to a gain of KATP channel function , which will attenuate the β-cell response to increased metabolism and will thereby decrease insulin secretion . (86)Rb(+) flux assays on COSm6 cells coexpressing the mutant subunits ( to recapitulate the compound heterozygous state ) show a 2-fold increase in basal rate of (86)Rb(+) efflux relative to WT channels . Experiments on excised inside-out patches also reveal a slight increase in activity , manifested as an enhancement in stimulation by MgADP in channels expressing the compound heterozygous mutations or homozygous G296R mutation . In addition , the IC 50 for DB00171 inhibition of homomeric A30V channels was increased ~6-fold , and was increased ~3-fold for both heteromeric A30V+WT channels or compound heterozygous ( A30V +G296R ) channels . Thus , each mutation makes a mechanistically distinct contribution to the channel gain-of-function that results in neonatal diabetes , and which we predict may contribute to diabetes in related carrier individuals . Displacement of Q01826 -bound histone deacetylase 1 corepressor by the human immunodeficiency virus type 1 transactivator induces expression of interleukin-2 and its receptor in T cells . One hallmark of human immunodeficiency virus type 1 ( HIV-1 ) infection is the dysregulation of cytokine gene expression in T cells . Transfection of T cells with human T-cell leukemia type 1 or 2 transactivator results in the induction of the T-cell-restricted cytokine interleukin-2 ( P60568 ) and its receptor ( IL-2Ralpha ) . However , no T-cell-specific factor(s) has been directly linked with the regulation of P60568 and IL-2Ralpha transcription by influencing the promoter activity . Thymocytes from Q01826 ( special AT-rich sequence binding protein 1 ) knockout mice have been shown to ectopically express IL-2Ralpha , suggesting involvement of Q01826 in its negative regulation . Here we show that Q01826 , a T-cell-specific global gene regulator , binds to the promoters of human P60568 and IL-2Ralpha and recruits histone deacetylase 1 ( Q13547 ) in vivo . Q01826 also interacts with Tat in HIV-1-infected T cells . The functional interaction between HIV-1 Tat and Q01826 requires its PDZ-like domain , and the binding of the Q13547 corepressor occurs through the same . Furthermore , Tat competitively displaces Q13547 that is bound to Q01826 , leading to increased acetylation of the promoters in vivo . Transduction with Q01826 interaction-deficient soluble Tat ( Tat 40-72 ) and reporter assays using a transactivation-negative mutant ( C22G ) of Tat unequivocally demonstrated that the displacement of Q13547 itself is sufficient for derepression of these promoters in vivo . These results suggest a novel mechanism by which HIV-1 Tat might overcome Q01826 -mediated repression in T cells . Stereospecific interaction of a novel spirosuccinimide type aldose reductase inhibitor , DB05327 , with aldose reductase . P15121 ( AR ) is an NADPH-dependent enzyme implicated in diabetic complications . DB05327 [ ( R ) -(-)-2-(4-bromo-2-fluorobenzyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-4-spiro-3'-pyrrolidine-1,2',3,5'-tetrone ] is a structurally novel and potent Q9Y4X5 with an inhibitor constant ( K(i) = 10(-)(10) M ) 2000-fold lower than that of its optical antipode ( S-isomer ) . To elucidate the inhibition modes and the stereochemical differences in their inhibitory potencies , we examined the interaction of these R- and S-isomers with AR under physiological conditions . Enzyme kinetic analysis , which was performed by using physiological substrates at 37 degrees C , showed that both isomers selectively act on the E-NADP(+) complex in both the forward and reverse reactions of AR . However , fluorometric titration analysis demonstrated that the affinities of the isomers for the E-NADP(+) complex are about the same as those for the E-NADPH complex and the apoenzyme . These results suggested that the selective binding to the E-NADP(+) complex arises from the predominance of this enzyme form during steady-state turnover rather than from binding specificity . Both the competition with a known active site-directed Q9Y4X5 and the protective effect on AR inactivation by N-bromosuccinimide showed that the isomers bind to the active site of the enzyme , but the thermodynamic parameters for the binding to AR indicated that additional hydrogen bonds and/or van der Waals interactions contribute to the energetic stabilization in the E-R-isomer complex . Molecular modeling , together with the deductions from spectroscopic studies , suggested that the succinimide ring and the 4-bromo-2-fluorobenzyl group of the R-isomer are optimally located for formation of a hydrogen-bonding network with AR , and that the latter benzyl group is also effective for the differentiation between AR and aldehyde reductase ( a closely related enzyme ) . DB00428 -nicotinamide-induced diabetes in the rat . Characteristics of the experimental model . Administration of both streptozotocin ( Q11206 ) and nicotinamide ( NA ) has been proposed to induce experimental diabetes in the rat . Q11206 is well known to cause pancreatic B-cell damage , whereas NA is administered to rats to partially protect insulin-secreting cells against Q11206 . Q11206 is transported into B-cells via the glucose transporter P11168 and causes DNA damage leading to increased activity of poly(ADP-ribose) polymerase ( P09874 ) to repair DNA . However , exaggerated activity of this enzyme results in depletion of intracellular NAD(+) and DB00171 , and the insulin-secreting cells undergo necrosis . The protective action of NA is due to the inhibition of P09874 activity . NA inhibits this enzyme , preventing depletion of NAD(+) and DB00171 in cells exposed to Q11206 . Moreover , NA serves as a precursor of NAD(+) and thereby additionally increases intracellular NAD(+) levels . The severity of diabetes in experimental rats strongly depends on the doses of Q11206 and NA given to these animals . Therefore , in diabetic rats , blood glucose may be changed in a broad range -- from slight hyperglycemia to substantial hyperglycemia compared with control animals . Similarly , blood insulin may be only slightly decreased or substantial hypoinsulinemia may be induced . In vitro studies demonstrated that the insulin-secretory response to glucose is attenuated in Q11206 -NA-induced diabetic rats compared with control animals . This is due to reduced B-cell mass as well as metabolic defects in the insulin-secreting cells . Results of numerous experiments have demonstrated that this model of diabetes is useful in studies of different aspects of diabetes . Novel phenolic antioxidants as multifunctional inhibitors of inducible P19320 expression for use in atherosclerosis . A series of novel phenolic compounds has been discovered as potent inhibitors of P01375 -inducible expression of vascular cell adhesion molecule-1 ( P19320 ) with concurrent antioxidant and lipid-modulating properties . Optimization of these multifunctional agents led to the identification of 3a ( DB05399 ) as a clinical candidate with demonstrated efficacies in animal models of atherosclerosis and hyperlipidemia .

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interacts_with DB01016?

multiple_choice

DB00898 blocks leukocyte recruitment and endothelial cell activation in vivo and in vitro . Immune system impairment and increased susceptibility to infection among alcohol abusers is a significant but not well-understood problem . We hypothesized that acute ethanol administration would inhibit leukocyte recruitment and endothelial cell activation during inflammation and infection . Using LPS and carrageenan air pouch models in mice , we found that physiological concentrations of ethanol ( 1-5 g/kg ) significantly blocked leukocyte recruitment ( 50-90 % ) . Because endothelial cell activation and immune cell-endothelial cell interactions are critical regulators of leukocyte recruitment , we analyzed the effect of acute ethanol exposure on endothelial cell activation in vivo using the localized Shwartzman reaction model . In this model , ethanol markedly suppressed leukocyte accumulation and endothelial cell adhesion molecule expression in a dose-dependent manner . Finally , we examined the direct effects of ethanol on endothelial cell activation and leukocyte-endothelial cell interactions in vitro . DB00898 , at concentrations within the range found in human blood after acute exposure and below the levels that induce cytotoxicity ( 0.1-0.5 % ) , did not induce endothelial cell activation , but significantly inhibited P01375 -mediated endothelial cell activation , as measured by adhesion molecule ( P16581 , P05362 , P19320 ) expression and chemokine ( P10145 , P13500 , RANTES ) production and leukocyte adhesion in vitro . Studies exploring the potential mechanism by which ethanol suppresses endothelial cell activation revealed that ethanol blocked NF-kappaB nuclear entry in an P25963 -dependent manner . These findings support the hypothesis that acute ethanol overexposure may increase the risk of infection and inhibit the host inflammatory response , in part , by blocking endothelial cell activation and subsequent immune cell-endothelial cell interactions required for efficient immune cell recruitment . DB03203 1-phosphate inhibits nitric oxide production induced by interleukin-1beta in rat vascular smooth muscle cells . DB03203 1-phosphate ( Q14703 ) is a lipid mediator that exerts potent and diverse biological effects on several cardiovascular cells . We investigated the effect of Q14703 on interleukin ( IL ) -1beta-induced nitric oxide ( NO ) production and inducible NO synthase ( P35228 ) expression in rat vascular smooth muscle cells ( VSMCs ) . Q14703 inhibited NO production at concentrations higher than 0.1 muM ; this was associated with the inhibition of P35228 protein and mRNA expression . Q14703 also inhibited IL-1beta-induced P30793 ( GTPCH ) mRNA expression . Pertussis toxin ( PTX ) partially attenuated the inhibitory effects of Q14703 on NO production and P35228 protein induction , whereas it completely blocked the inhibitory effects on P35228 and GTPCH mRNA expression . Q14703 inhibited P35228 expression in Ca(2+)-depleted conditions ; PTX did not modify this effect . The Rho kinase inhibitor Y 27632 partially but significantly attenuated the inhibitory effect of Q14703 on P35228 expression in Ca(2+)-depleted condition but did not affect it in the presence of Ca(2+) . Q14703 significantly inhibited IL-1beta-induced persistent activation of extracellular signal-regulated kinase ( P29323 ) but had no effect in Ca(2+)-depleted conditions . Thus , Q14703 inhibits IL-1beta induction of NO production and P35228 expression in rat VSMCs through multiple mechanisms involving both PTX-sensitive and -insensitive G proteins coupled to Q14703 receptors . Furthermore , Ca(2+)-dependent P29323 inhibition and Ca(2+)-independent Rho kinase activation might be involved in the inhibitory mechanism of P35228 expression . Through its action on NO production by VSMCs , Q14703 may play an important role in the progression of local vascular injury associated with thrombosis , atherosclerosis , and hypertension . Enforced expression of GATA-3 in transgenic mice inhibits Th1 differentiation and induces the formation of a T1/ST2-expressing Th2-committed T cell compartment in vivo . The transcription factor GATA-3 is essential for early T cell development and differentiation of naive P01730 (+) T cells into Th2 effector cells . To study the function of GATA-3 during T cell-mediated immune responses in vivo , we investigated P06729 - P23771 -transgenic mice in which GATA-3 expression is driven by the P06729 locus control region . Both in the P01730 (+) and the CD8(+) T cell population the proportion of cells exhibiting a P16070 (high)CD45RB(low)CD62L(low) Ag-experienced phenotype was increased . In P06729 - P23771 -transgenic mice , large fractions of peripheral P01730 (+) T cells expressed the IL-1 receptor family member T1/ST2 , indicative of advanced Th2 commitment . Upon in vitro T cell stimulation , the ability to produce P60568 and P01579 was decreased . Moreover , P01730 (+) T cells manifested rapid secretion of the Th2 cytokines P05112 , P05113 , and P22301 , reminiscent of Th2 memory cells . In contrast to wild-type P01730 (+) cells , which lost GATA-3 expression when cultured under Th1-polarizing conditions , P06729 - P23771 -transgenic P01730 (+) cells maintained expression of GATA-3 protein . Under Th1 conditions , cellular proliferation of P06729 - P23771 -transgenic P01730 (+) cells was severely hampered , P01579 production was decreased and Th2 cytokine production was increased . Enforced GATA-3 expression inhibited Th1-mediated in vivo responses , such as Ag-specific IgG2a production or a delayed-type hypersensitivity response to DB05299 . Collectively , these observations indicate that enforced GATA-3 expression selectively inhibits Th1 differentiation and induces Th2 differentiation . The increased functional capacity to secrete Th2 cytokines , along with the increased expression of surface markers for Ag-experienced Th2-committed cells , would argue for a role of GATA-3 in Th2 memory formation . P30047 -dependent and -independent inhibitors of P30793 . P30047 ( P30047 ) mediates the feedback inhibition of P30793 activity by ( 6R ) -L-erythro- DB00360 ( BH4 ) through protein complex formation . Since guanine and BH4 have a common pyrimidine ring structure , we examined the inhibitory effect of guanine and its analogs on the enzyme activity . DB02377 , 8-hydroxyguanine , 8-methylguanine , and 8-bromoguanine inhibited the enzyme activity in a P30047 -dependent and pH-dependent manner and induced complex formation between P30793 and P30047 . The type of inhibition by this group is a mixed type . All these properties were shared with BH4 . In striking contrast , inhibition by DB01667 and 8-mercaptoguanine was P30047 -independent and pH-independent . The type of inhibition by DB01667 and 8-mercaptoguanine was a competitive type . The two compounds did not induce complex formation between the enzyme and P30047 . These results demonstrate that guanine compounds of the first group bind to the BH4-binding site of the P30793 / P30047 complex , whereas DB01667 and 8-mercaptoguanine bind to the active site of the enzyme . Finally , the possible implications in Lesch-Nyhan syndrome and Parkinson diseases of the inhibition of P30793 by guanine and 8-hydroxyguanine are discussed . DB01016 -induced apoptosis is specifically enhanced by expression of the sulfonylurea receptor isoform Q09428 but not by expression of SUR2B or the mutant Q09428 (M1289T) . Q09428 ( Q09428 ) is the regulatory subunit of the pancreatic DB00171 -sensitive K+ channel ( K( DB00171 ) channel ) , which is essential for triggering insulin secretion via membrane depolarization . Sulfonylureas , such as glibenclamide and tolbutamide , act as K( DB00171 ) channel blockers and are widely used in diabetes treatment . These antidiabetic substances are known to induce apoptosis in pancreatic beta-cells or beta-cell lines under certain conditions . However , the precise molecular mechanisms of this sulfonylurea-induced apoptosis are still unidentified . To investigate the role of Q09428 in apoptosis induction , we tested the effect of glibenclamide on recombinant human embryonic kidney 293 cells expressing either Q09428 , the smooth muscular isoform SUR2B , or the mutant Q09428 (M1289T) at which a single amino acid in transmembrane helix 17 ( TM17 ) was exchanged by the corresponding amino acid of SUR2 . By analyzing cell detachment , nuclear condensation , DNA fragmentation , and caspase-3-like activity , we observed a Q09428 -specific enhancement of glibenclamide-induced apoptosis that was not seen in SUR2B , Q09428 (M1289T) , or control cells . Coexpression with the pore-forming Kir6.2 subunit did not significantly alter the apoptotic effect of glibenclamide on Q09428 cells . In conclusion , expression of Q09428 , but not of SUR2B or Q09428 (M1289T) , renders cells more susceptible to glibenclamide-induced apoptosis . Therefore , Q09428 as a pancreatic protein could be involved in specific variation of beta-cell mass and might also contribute to the regulation of insulin secretion at this level . According to our results , TM17 is essentially involved in Q09428 -mediated apoptosis . This effect does not require the presence of functional Kir6.2-containing K( DB00171 ) channels , which points to additional , so far unknown functions of Q09428 . P13726 -independent effects of recombinant factor VIIa on hemostasis . The molecular mechanisms responsible for the hemostatic efficacy of recombinant activated factor VII ( DB00036 ; NovoSeven , Novo Nordisk , Bagsvaerd , Denmark ) in platelet-related bleeding disorders remain unclear . The general concept is that DB00036 locally enhances thrombin generation at the site of injury , where tissue factor ( TF ) has become exposed . However , a growing amount of evidence shows that DB00036 is also able to exert its activity in a manner independent of TF . Using an in vitro flow model , we recently showed that TF-independent thrombin generation is responsible for increased platelet deposition onto injured vessels following DB00036 administration . Furthermore , it has been shown that DB00036 can restore platelet aggregation in Glanzmann 's thrombasthenia ( GT ) patients via TF-independent thrombin generation . However , the mechanism behind TF-independent thrombin generation remains to be elucidated . It is postulated that , in vivo , both the TF-dependent and TF-independent thrombin generation induced by DB00036 contribute to the control of hemorrhage in patients with platelet-related bleeding disorders and , perhaps , other causes of hemorrhagic diatheses . Involvement of the mitogen-activated protein kinase c-Jun NH2-terminal kinase 1 in thrombus formation . The involvement of the mitogen-activated protein kinase c-Jun NH2-terminal kinase-1 ( P45983 ) has never been investigated in hemostasis and thrombosis . Using two JNK inhibitors ( SP600125 and 6o ) , we have demonstrated that P45983 is involved in collagen-induced platelet aggregation dependent on ADP . In these conditions , P45983 activation requires the coordinated signaling pathways of collagen receptors ( alpha2beta1 and glycoprotein (GP)VI ) and ADP . In contrast , P45983 is not required for platelet adhesion on a collagen matrix in static or blood flow conditions ( 300-1500 s(-1) ) involving collagen receptors ( alpha2beta1 and Q9HCN6 ) . Importantly , at 1500 s(-1) , P45983 acts on thrombus formation on a collagen matrix dependent on GPIb- P04275 ( P04275 ) interaction but not ADP receptor activation . This is confirmed by the involvement of P45983 in shear-induced platelet aggregation at 4000 s(-1) . We also provide evidence during rolling and adhesion of platelets to P04275 that platelet GPIb- P04275 interaction triggers alphaIIbbeta3 activation in a P45983 -dependent manner . This was confirmed with a Glanzmann thrombastenic patient lacking alphaIIbbeta3 . Finally , in vivo , P45983 is involved in arterial but not in venular thrombosis in mice . Overall , our in vitro studies define a new role of P45983 in thrombus formation in flowing blood that is relevant to thrombus development in vivo . A protective role of hydrogen sulfide against oxidative stress in rat gastric mucosal epithelium . We investigated effect of hydrogen sulfide ( H(2)S ) on oxidative stress-caused cell death in gastric mucosal epithelial cells . In rat normal gastric epithelial RGM1 cells , NaHS , a H(2)S donor , at 1.5mM strongly suppressed hydrogen peroxide ( H(2)O(2) ) -caused cell death , while it slightly augmented the H(2)O(2) toxicity at 0.5-1mM . The protective effect of NaHS was abolished by inhibitors of MEK or JNK , but not of p38 Q96HU1 kinase . NaHS at 1.5mM actually phosphorylated P29323 and JNK in RGM1 cells . DB01016 , an DB00171 -sensitive K(+) ( K( DB00171 )(+) ) channel inhibitor , did not affect the protective effect of NaHS , although mRNAs for K( DB00171 )(+) channel subunits , Kir6.1 and Q09428 , were detected in RGM1 cells . In anesthetized rats , oral administration of NaHS protected against gastric mucosal lesion caused by ischemia-reperfusion . These results suggest that NaHS/H(2)S may protect gastric mucosal epithelial cells against oxidative stress through stimulation of Q96HU1 kinase pathways , a therapeutic dose range being very narrow . Met326Ile aminoacid polymorphism in the human p85 alpha gene has no major impact on early insulin signaling in type 2 diabetes . Class I alpha phosphatidylinositol ( PI ) 3-kinase is an important enzyme in the early insulin signaling cascade , and plays a key role in insulin-mediated glucose transport . Despite extensive investigation , the genes responsible for the development of the common forms of type 2 diabetes remain unknown . This study was performed to identify variants in the coding region of p85 alpha , the regulatory subunit of PI 3-kinase . Fibroblasts from skin biopsies from type 2 diabetics and controls were established to address this issue . P85 alpha cDNA was sequenced , and a single point mutation at codon 326 was found . This mutation resulted in a homozygous missense amino acid change DB00134 --> DB00167 in one subject with type 2 diabetes and heterozygous variant in two other diabetic patients and one with severe insulin resistance . Interestingly , those patients revealed an impaired insulin-mediated insulin receptor substrate ( P41252 ) -1 binding to p85 alpha without any alteration in Q9Y4H2 /p85 alpha association . Furthermore , P35568 , Q9Y4H2 , p85 alpha and MAPK protein contents were not significantly changed , and neither were MAPK or Akt phosphorylation . We conclude from our data that this variant may have only minor impact on signaling events ; however , in combination with variants in other genes encoding signaling proteins , this may have a functional impact on early insulin signaling . P07550 mediated P29323 activation is regulated by interaction with Q5TCQ9 . The beta-2 adrenergic receptor ( beta2AR ) has a carboxyl terminus motif that can interact with P78352 /discs-large/ Q07157 homology ( PDZ ) domain-containing proteins . In this paper , we identified membrane-associated guanylate kinase inverted-3 ( Q5TCQ9 ) as a novel binding partner of beta2AR . The carboxyl terminus of beta2AR binds with high affinity to the fifth PDZ domain of Q5TCQ9 , with the last four amino acids ( D-S-L-L ) of the receptor being the key determinants of the interaction . In cells , the association of full-length beta2AR with Q5TCQ9 occurs constitutively and is enhanced by agonist stimulation of the receptor . Our data also demonstrated that beta2AR-stimulated extracellular signal-regulated kinase-1/2 ( P27361 /2 ) activation was substantially retarded by Q5TCQ9 expression . These data suggest that Q5TCQ9 regulates beta2AR-mediated P29323 activation through the physical interaction between beta2AR and Q5TCQ9 . DB01016 exerts an antitumor activity through reactive oxygen species-c-jun NH2-terminal kinase pathway in human gastric cancer cell line MGC-803 . DB01016 , a blocker of DB00171 -sensitive potassium ( K( DB00171 ) ) channels , can suppress progression of many cancers , but the involved mechanism is unclear . Herein we reported that MGC-803 cells expressed the K( DB00171 ) channels composed of Kir6.2 and Q09428 subunits . DB01016 induced cellular viability decline , coupled with cell apoptosis and reactive oxygen species ( ROS ) generation in MGC-803 cells . Meanwhile , glibenclamide increased NADPH oxidase catalytic subunit gp91(phox) expression and superoxide anion ( O2- ) generation , and caused mitochondrial respiration dysfunction in MGC-803 cells , suggesting that glibenclamide induced an increase of ROS derived from NADPH oxidase and mitochondria . DB01016 could also lead to loss of mitochondrial membrane potential , release of cytochrome c and apoptosis-inducing factor ( O95831 ) , and activation of c-jun NH2-terminal kinase ( JNK ) in MGC-803 cells . Pretreatment with antioxidant N-acetyl-l-cysteine ( Q9C000 ) prevented glibenclamide-induced JNK activation , apoptosis and cellular viability decline . Furthermore , glibenclamide greatly decreased the cellular viability , induced apoptosis and inhibited Akt activation in wild-type mouse embryonic fibroblast ( MEF ) cells but not in P45983 -/- or P45984 -/- MEF cells . Taken together , our study reveals that glibenclamide exerts an antitumor activity in MGC-803 cells by activating ROS-dependent , JNK-driven cell apoptosis . These findings provide insights into the use of glibenclamide in the treatment of human gastric cancer . Influence of genetic polymorphisms on the pharmacokinetics and pharmaco-dynamics of sulfonylurea drugs . Sulfonylurea drugs including chlorpropamide , gliclazide , tolbutamide , glipizide , glibenclamide ( glyburide ) and glimepiride are the most widely used oral hypoglycaemic agents in people with type 2 diabetes . This review investigates the impact of genetic polymorphisms on the pharmacokinetics and pharmacodynamics of sulfonylurea drugs . P11712 is the major enzyme involved in sulfonylurea drug metabolism . P11712 variant allele carriers have significant lower apparent clearance of these medicines . P33261 genotype is more influential for gliclazide pharmacokinetics when compared to P11712 . Sulfonylurea pharmacodynamics is affected by several genes . Q09428 ( Q09428 , Q09428 gene ) and K+ inward rectifier Kir6.2 ( Q14654 ) have been correlated to significant variation in sulfonylurea response . Diabetics with the Q09428 exon 33 G allele are more sensitive to gliclazide and the rs5210 variant of the Q14654 gene was associated with improved clinical efficacy of gliclazide . Carriers of Q9NQB0 ( Q9NQB0 ) variants are more likely to fail sulfonylurea therapy . On the other hand , patients with HNF-1alpha mutations had a significant greater response to gliclazide when compared to those with type 2 diabetes . The Arg972 polymorphism of insulin receptor substrate 1 ( P35568 ) may lead to secondary failure of sulfonylurea therapy . Calpain 10 gene ( Q9HC96 ) polymorphism has also been linked to sulfonylurea drug response . Despite the available evidence , larger population studies that investigate the pharmacokinetics and pharmacodynamics of sulfonylurea drugs are needed to investigate the influence of key SNPs amidst all potential contributing factors to variability in response to these which inturn will provide information to optimise sulfonylurea use in people with diabetes . Ras-dependent P29323 activation by the human G(s)-coupled serotonin receptors Q13639 (b) and P34969 (a) . Receptor tyrosine kinases activate mitogen-activated protein ( Q96HU1 ) kinases through Ras , P04049 , and MEK . Receptor tyrosine kinases can be transactivated by G protein-coupled receptors coupling to G(i) and G(q) . The human G protein-coupled serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) couple to G(s) and elevate intracellular DB02527 . Certain G(s)-coupled receptors have been shown to activate Q96HU1 kinases through a protein kinase A- and Rap1-dependent pathway . We report the activation of the extracellular signal-regulated kinases ( ERKs ) 1 and 2 ( Q8TCB0 and Q8NFH3 Q96HU1 kinase ) through the human serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) in COS-7 and human embryonic kidney HEK293 cells . In transfected HEK293 cells , 5-HT-induced activation of P27361 /2 is sensitive to H89 , which indicates a role for protein kinase A . The observed activation of P27361 /2 does not require transactivation of epidermal growth factor receptors . Furthermore , 5-HT induced activation of both Ras and Rap1 . Whereas the presence of P47736 did not influence the 5-HT-mediated activation of P27361 /2 , the activation of P27361 /2 was abolished in the presence of dominant negative Ras ( RasN17 ) . P27361 /2 activation was reduced in the presence of " dominant negative " Raf1 ( RafS621A ) and slightly reduced by dominant negative B-Raf , indicating the involvement of one or more Raf isoforms . These findings suggest that activation of P27361 /2 through the human G(s)-coupled serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) in HEK293 cells is dependent on Ras , but independent of Rap1 . Pharmacological treatment of irritable bowel syndrome -- from concept to sales . Functional gastrointestinal disorders are characterised by central and peripheral physiological changes , associated with psychological factors . Successful drug development has been hindered by lack of adequate characterisation of the nature of symptoms and their physiological and psychological correlates . Animal models of chronic stress are lacking . High levels of drug safety are now demanded for treating non-life threatening conditions . Once close to market , patient pressure groups , health care providers and insurers , government , and the internet can all influence a drug 's success . Serotonin-modifying drugs have been the main recent focus of development , with mixed results . Cisapride has been withdrawn because of concerns related to QT prolongation and cardiac arrhythmias . The 5- Q9H205 antagonists have been developed on the questionable assumption that they modify visceral sensation in patients . Problems have arisen with alosetron being associated with ischaemic colitis and a high incidence of constipation . The Q13639 agonists have their major effect on inducing peristalsis , and may modify gut secretion and sensory function . DB01079 and prucalopride show promise in patients with constipation and related symptoms . 5-HT1 agonists may play a role in treating functional dyspepsia , partly by improving impaired gastric accommodation to a meal . Antidepressants , often found to be clinically beneficial in these disorders , also affect serotonin metabolism . Past successes , such as loperamide or the somatostatin analogue octreotide , involved targeting end organ receptors influencing motor function or secretion . Modifying sensory function is much more challenging . Future research with novel compounds need to keep these recent lessons in mind . The oncogenic P41212 / P09619 beta fusion protein induces cell death through JNK/SAPK pathway . The P41212 / P09619 beta ( T/P ) fusion protein isolated from patients bearing a t(5;12) translocation is transforming when expressed in haematopoietic cells . To examine the signal transduction events activated by this protein , we measured the effect of T/P on activation of the c-Jun N-terminal kinase/stress-activated protein kinase ( JNK/SAPK ) in mouse bone marrow-derived Ba/ P13726 cells . Significant increase in the activity of JNK/ P45983 was observed in transient transfection as well as in Ba/ P13726 cells stably expressing T/P . This activation was abrogated when the T/P-expressing cells were treated with a specific inhibitor of the P09619 beta tyrosine kinase , indicating that the activity of the P09619 beta part of the fusion protein was involved in JNK/SAPK activation . Expression of a dominant negative mutant of mitogen-activated protein kinase kinase 4 ( P45985 ) , a direct activator of JNK/SAPK , prevented T/P-induced JNK/SAPK activation . In addition , inhibition of phosphoinositide-3 OH kinase ( P19957 kinase ) , a promoting survival factor , potentiated the effect of T/P on JNK/SAPK activation . Interestingly , expression of T/P was shown to initiate an apoptotic response that was enhanced by treatment of cells with the P19957 kinase inhibitor LY294002 , suggesting that T/P mediated cell death through activation of JNK/SAPK signalling pathway . Consistent with this hypothesis , expression of the dominant negative mutant of P45985 decreased T/P-mediated apoptosis , while a dominant-negative mutant of P19957 kinase enhances cell death . These findings indicate that activation of JNK/SAPK by T/P is related to apoptosis rather than cell proliferation and transformation . Inhibitors of DB00171 -binding cassette transporters suppress interleukin-12 p40 production and major histocompatibility complex II up-regulation in macrophages . DB00171 -binding cassette ( DB01048 ) transporters are a large family of proteins whose role is to translocate various substances across biological membranes . They include the Tangier disease protein ABC1 , sulfonylurea receptors ( Q09428 ) , multidrug resistance protein ( MDR ) , and cystic fibrosis transmembrane regulator ( P13569 ) . In the current study , we investigated the involvement of ABC transporters in the regulation of lipopolysaccharide ( LPS ) and/or interferon ( IFN ) -gamma-induced interleukin ( IL ) -12 p40 and tumor necrosis factor ( P01375 ) -alpha production , nitric oxide formation , as well as major histocompatibility complex II up-regulation in macrophages . The general ABC transporter inhibitor glibenclamide suppressed both IL-12 p40 and nitric oxide production . However , glibenclamide failed to affect the production of P01375 . The selective ABC1 inhibitors 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid and sulfobromophthalein mimicked the suppressive effect of glibenclamide on IL-12 p40 production . On the other hand , both the MDR inhibitor verapamil and P13569 blocker 2,2'-iminodibenzoic acid failed to suppress the production of IL-12 p40 . Furthermore , selective inhibitors and activators of SURs were without effect . In agreement with the pharmacological data , macrophages expressed mRNA for ABC1 , but not SURs or P13569 . Intracellular levels of IL-12 p40 were decreased by glibenclamide , suggesting that glibenclamide does not affect IL-12 p40 secretion . The effect of glibenclamide did not involve an interference with the activation of the p38 and Q8NFH3 /44 mitogen-activated protein kinases or c-Jun kinase . DB01016 also suppressed P01579 -induced up-regulation of major histocompatibility complex II . Taken together , our results indicate that ABC proteins regulate LPS and/or P01579 -induced macrophage activation . Inhibition of P04275 -mediated platelet activation and thrombosis by the anti- P04275 A1-domain aptamer DB05202 . BACKGROUND : P04275 ( P04275 ) has a role in both hemostasis and thrombosis . Platelets adhere to damaged arteries by interactions between the P04275 A1-domain and glycoprotein Ib receptors under conditions of high shear . This initial platelet binding event stimulates platelet activation , recruitment , and activation of the clotting cascade , promoting thrombus formation . OBJECTIVE : To characterize the inhibitory activity of a P04275 inhibitory aptamer . METHODS : Using in vitro selection , aptamer stabilization , and conjugation to a 20-kDa poly ( ethylene glycol ) , we generated a nuclease-resistant aptamer , DB05202 , that binds to the P04275 A1-domain with high affinity ( K(D) approximately 2 nM ) . The aptamer was assessed for inhibition of P04275 -induced platelet aggregation . In vitro inhibition of platelet adhesion was assessed on collagen-coated slides and injured pig aortic segments . In vivo activity was assessed in a cynomolgus monkey carotid electrical injury thrombosis model . RESULTS AND CONCLUSION : DB05202 inhibited botrocetin-induced platelet aggregation ( IC90 approximately 300 nM ) and shear force-induced platelet aggregation ( IC95 approximately 400 nM ) . It reduced adhesion of platelets to collagen-coated matrices and formation of platelet thrombi on denuded porcine arteries . DB05202 also inhibited the formation of occlusive thrombi in cynomolgus monkeys . We have discovered a novel anti- P04275 aptamer that could have therapeutic use as an anti- P04275 agent in the setting of P04275 -mediated thrombosis . New concepts in anti-tumor necrosis factor therapy for inflammatory bowel disease . Crohn 's disease is a T helper type 1 response immune disease characterized by increased production of interleukin-12 tumor necrosis factor-a ( P01375 ) , and interferon-g . Clinical trials have demonstrated that inhibition of P01375 is effective for the treatment of Crohn 's disease . Adverse events reported in patients treated with anti- P01375 agents include immunogenicity , acute infusion reactions , delayed hypersensitivity-type reactions , autoimmune diseases including drug-induced lupus and demyelination , and infection . This article reviews new concepts in the treatment of Crohn 's disease and ulcerative colitis with a variety of anti- P01375 biologic therapies : infliximab , adalimumab , DB08904 , CDP571 , etanercept , and onercept . P22309 *28 is associated with greater decrease in serum K⁺ levels following oral intake of procaterol . BACKGROUND AND OBJECTIVE : DB01366 is a potent β2-agonist frequently used for the management of asthma and chronic obstructive pulmonary disease . The efficacy and adverse effects of β2-agonists are heterogeneous in individual patients , which may be partly caused by genetic variations in metabolizing enzymes and receptor molecules . The present study was designed to analyze the relationship between gene polymorphisms and physiological effects of procaterol in healthy subjects . METHODS : Ninety-two non-smoking healthy volunteers were given 1 µg/kg body weight ( max 50 µg ) of procaterol as a dry syrup preparation , and the serum concentrations of procaterol , serum K(+) , and the physical responses were monitored for 240 min . We genotyped β2-adrenergic receptor ( P07550 ) ( Arg16Gly and Gln27Glu ) , cytochrome P450 3A4 ( rs2246709 , rs4646437 ) , and uridine diphosphate glucuronosyltransferase 1A1 ( P22309 ) ( rs4148323 [ allele A , *6 ] , rs12479045 , rs4148328 , rs4663971 , rs12052787 , rs4148329 , A (TA)6/7 TAA [ seven-repeat allele , *28 ] ) . DB01366 concentrations in serum were measured by liquid chromatography-tandem mass spectrometry . RESULTS : No gene polymorphisms affected serum procaterol concentrations . Meanwhile , overall serum K(+) level changes were significantly lower in carriers of P22309 *28 than in non-carriers after correcting for strong effects of serum procaterol concentrations and baseline K(+) levels . No other polymorphisms were associated with serum K(+) levels . None of polymorphisms of P07550 were associated with any physical responses . CONCLUSION : The present study indicates that significant hypokalemia may occur in carriers of P22309 *28 by systemic administration of procaterol and potentially by other β2-agonists metabolized in the liver .

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interacts_with DB01296?

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DB06366 for the treatment of human epidermal growth factor receptor type 2-positive metastatic breast cancer . PURPOSE : The pharmacology , pharmacokinetics , clinical efficacy , safety , and administration of pertuzumab in patients with metastatic human epidermal growth factor receptor type 2 ( P04626 ) -positive breast cancer are reviewed . SUMMARY : Disease progression in P04626 -positive breast cancer is often due to resistance to or a lack of efficacy of trastuzumab-based anti- P04626 therapy . DB06366 is the first humanized monoclonal antibody in a new class of drugs , the HER dimerization inhibitors , approved by the Food and Drug Administration for the first-line treatment of patients with metastatic P04626 -positive breast cancer who have not received prior anti- P04626 therapy or chemotherapy for metastatic disease . Since pertuzumab binds to a different epitope than trastuzumab , combination therapy with pertuzumab and trastuzumab results in a more complete blockade of P04626 signaling than trastuzumab monotherapy . The efficacy of adding pertuzumab to trastuzumab-docetaxel dual therapy was demonstrated in a pivotal randomized multicenter Phase III trial , which showed a significant benefit in terms of progression-free survival , with improved overall survival , in favor of the triple therapy as an initial regimen in treatment-naive patients with metastatic P04626 -positive breast cancer . The combination of pertuzumab and trastuzumab has been found to have a tolerable toxicity profile . As clinical trials of pertuzumab for adjuvant , neoadjuvant , and metastatic-disease treatment continue , its role in the treatment of P04626 -positive breast cancer will continue to evolve . CONCLUSION : DB06366 , a novel P04626 dimerization inhibitor , has been shown to be effective in the treatment of metastatic P04626 -positive breast cancer when used in combination with trastuzumab and docetaxel and is recommended for first-line therapy . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . Fyn is a novel target of DB03823 gallate in the inhibition of JB6 Cl41 cell transformation . The cancer preventive action of DB03823 gallate ( EGCG ) , found in green tea , is strongly supported by epidemiology and laboratory research data . However , the mechanism by which EGCG inhibits carcinogenesis and cell transformation is not clear . In this study , we report that EGCG suppressed epidermal growth factor ( P01133 ) -induced cell transformation in JB6 cells . We also found that EGCG inhibited P01133 -induced Fyn kinase activity and phosphorylation in vitro and in vivo . Fyn was implicated in the process because P01133 -induced JB6 cell transformation was inhibited by small interfering RNA ( siRNA ) -Fyn-JB6 cells . With an in vitro protein-binding assay , we found that EGCG directly bound with the Q86UG4 -Fyn-SH2 domain but not the Q86UG4 -Fyn-SH3 domain . The K(d) value for EGCG binding to the Fyn SH2 domain was 0.367 +/- 0.122 microM and B(max) was 1.35 +/- 0.128 nmol/mg . Compared with control JB6 Cl41 cells , P01133 -induced phosphorylation of p38 Q96HU1 kinase ( p38 MAPK ) ( Thr180/Tyr182 ) , P39905 -2 ( Thr71 ) and signal transducer and activator of transcription 1 ( P42224 ) ( Thr727 ) was decreased in siRNA-Fyn-JB6 cells . EGCG could inhibit the phosphorylation of p38 MAPK , P39905 -2 , and P42224 . The DNA binding ability of AP-1 , P42224 , and P39905 -2 was also decreased in siRNA-Fyn-JB6 cells . Overall , these results demonstrated that EGCG interacted with Fyn and inhibited Fyn kinase activity and thereby regulated P01133 -induced cell transformation . Inhibition of Fyn kinase activity is a novel and important mechanism that may be involved in EGCG-induced inhibition of cell transformation . NT-702 ( parogrelil hydrochloride , DB05505 ) , a novel and potent phosphodiesterase inhibitor , improves reduced walking distance and lowered hindlimb plantar surface temperature in a rat experimental intermittent claudication model . NT-702 ( parogrelil hydrochloride , DB05505 ) , 4-bromo-6-[3-(4-chlorophenyl)propoxy]-5-[(pyridin-3-ylmethyl)amino]pyridazin-3(2H)-one hydrochloride , a novel phosphodiesterase ( PDE ) inhibitor synthesized as a potent vasodilatory and antiplatelet agent , is being developed for the treatment of intermittent claudication ( IC ) in patients with peripheral arterial disease . We assessed the efficacy of NT-702 in an experimental IC model as compared with cilostazol and additionally investigated the pharmacological property in vitro and ex vivo . NT-702 selectively inhibited PDE3 ( IC(50)=0.179 and 0.260 nM for Q14432 and 3B ) more potently than cilostazol ( IC(50)=231 and 237 nM for Q14432 and 3B ) among recombinant human PDE1 to PDE6 . NT-702 inhibited in vitro human platelet aggregation induced by various agonists ( IC(50)=11 to 67 nM ) and phenylephrine-induced rat aortic contraction ( IC(50)=24 nM ) . Corresponding results for cilostazol were 4.1 to 17 microM and 1.0 microM , respectively . NT-702 ( 3 mg/kg or more ) significantly inhibited ex vivo rat platelet aggregation after a single oral dose . For cilostazol , 300 mg/kg was effective . In a rat femoral artery ligation model , NT-702 at 5 and 10 mg/kg repeated oral doses twice a day ( P55957 ) for 13 days significantly improved the reduced walking distance while the lowered plantar surface temperature was improved at 2.5 mg/kg and more . DB01166 also improved the walking distance and surface temperature at 300 mg/kg P55957 but significant difference was only observed for surface temperature on day 8 . These results suggest that NT-702 can be expected to have therapeutic advantage for IC . [ Detection of point mutations of C-Ki-ras oncogene in colon cancer by PCR using specific oligonucleotide probes ] . Activation of C-Ki-ras oncogene by point mutation within codon 12,13 was determined by Polymerase Chain Reaction ( PCR ) using specific oligonucleotide probes . In 9 of 42 colon cancer specimens point mutations in codon 12 of C-Ki-ras oncogene were found . The point mutations identified were P19440 ( DB00145 ) ---- Q6IB77 ( DB00128 ) ( 4 cases ) , P19440 ( DB00145 ) ---- TGT ( DB00151 ) ( 3 cases ) and P19440 ( DB00145 ) ---- GTT( DB00161 ) ( 2 cases ) , respectively . Two types of point mutations ( P19440 ---- Q6IB77 , P19440 ---- TGT ) were found simultaneously in one specimen . The results showed that there was no relationship between the point mutation of C-Ki-ras oncogene and patient 's sex , age , state of metastasis or prognosis . DB00495 is effective against human multiple myeloma : a new use for an old drug ? DB00495 ( DB00495 ) is an antiretroviral drug that affects cell proliferation , apoptosis , and the NF-κB pathway . As multiple myeloma ( MM ) presents with constitutive activation of NF-κB , we analyzed the effect of DB00495 on human MM cell lines . We evaluated the cytotoxic effect of DB00495 in human MM cell lines sensitive ( 8226/S ) or resistant to doxorubicin ( 8226/DX5 ) and human T cell lymphoblast-like cells , uterine sarcoma cells , and HUVEC using MTT assay . Cytotoxicity was also evaluated in vivo in nude mice xenografted with 8226/S tumor . The effect of DB00495 on the expression of genes involved in cell proliferation , apoptosis , angiogenesis , and the NF-κB pathway was analyzed in the xenografts using real-time polymerase chain reaction . DB00495 was effective against both 8226/S and 8226/DX5 cells in a dose and time-dependent manner ( p = 0.02 ) in vitro and promoted cell cycle arrest in S phase in these cells . The tumor volume was lower in mice treated with DB00495 compared to untreated mice ( p = 0.0003 ) . DB00495 down-regulated the pro-proliferative genes encoding P31749 , MYC , P42224 , P45983 , P45984 , DB00833 -3 , Bcl-3 , and cyclin D2 ; pro-angiogenenic genes encoding P15692 and P10145 ; and genes involved in cell adhesion ( P05362 and P02751 ) and the NF-κB pathway . DB00495 up-regulated the expression of tumor suppressor gene Q9H334 and the pro-apoptotic genes encoding P55957 , O95999 , and caspase-8 . Thus , we demonstrated the cytotoxic effect of DB00495 in human MM cell lines for the first time . Our data may provide the rationale for future clinical trials of DB00495 for treating MM . Agents with selective estrogen receptor ( ER ) modulator activity induce apoptosis in vitro and in vivo in ER-negative glioma cells . Tamoxifen , a member of the selective estrogen receptor modulator ( SERM ) family , is widely used in the treatment of estrogen receptor ( ER ) -expressing breast cancer . It has previously been shown that high-dose tamoxifen has cytotoxic activity against glioma cells , but whether this effect is drug specific or represents a general property of SERMs is unknown . In this study , we demonstrate that tamoxifen and DB05487 , a novel benzopyranone with SERM activity , induce glioma cell apoptosis in a dose- and time-dependent manner . Moreover , administration of tamoxifen and DB05487 suppresses tumor growth in vivo and extends animal survival in glioma xenograft models . None of the eight glioma cell lines examined express either P03372 or -beta , suggesting the mechanism for tamoxifen- and DB05487 -induced glioma cell apoptosis is independent of the ER signaling pathway . Complementary DNA microarray expression profiling allowed us to identify a subset of genes specifically regulated by tamoxifen and DB05487 , and not by other apoptotic stimuli , including nuclear factor ( NF ) -kappaB with its target genes IEX-3 , P04179 , P05231 , and P10145 . We demonstrate that suppression of NF-kappaB activation markedly enhances SERM-induced apoptosis , suggesting a role for NF-kappaB in protecting glioma cells from SERM-induced cytotoxicity . These findings demonstrate for the first time that a SERM other than tamoxifen can induce glioma cell apoptosis in vitro and in vivo and that the clinical efficacy of SERMs for the treatment of malignant gliomas could potentially be enhanced by simultaneous inhibition of the NF-kappaB pathway . P35610 of the union : dynamics of distinct tumor-associated macrophage subsets governed by P42224 . The tumor stroma has long been ignored as therapeutic target , but it has become clear that several stromal cell types play a nonredundant role during tumor progression . In particular , macrophages possess the capacity to stimulate tumor growth and metastasis via multiple mechanisms . In this issue of the European Journal of Immunology , a study by Tymoszuk et al . Eur . J . Immunol . 2014 . 44 : 2247-2262 demonstrates that both monocyte recruitment and local macrophage proliferation determines the tumor-associated macrophage ( TAM ) pool size in P04626 /Neu-driven mammary carcinomas . These tumors contain two main TAM subsets -- MHC class II ( MHC-II ) (lo) F4/80(hi) and MHC-II(hi) F4/80(lo) -- similar to what was observed in other tumor models . Interestingly , only the MHC-II(lo) F4/80(hi) subset is largely absent in a P42224 -deficient background . P42224 induces the expression of P09603 , which in turn drives TAM proliferation and possibly also the M2 gene signature of MHC-II(lo) F4/80(hi) TAM . Conversely , P42224 deficiency upregulates M2 gene expression in MHC-II(hi) F4/80(lo) TAM , demonstrating that both TAM subsets are differentially regulated , probably as a consequence of their distinct intratumoral localization . In this Commentary , we place these findings in the context of current knowledge and propose new avenues for future research . Protein kinase C-associated kinase regulates NF-κB activation through inducing IKK activation . Activation of the transcription factor NF-κB induced by extracellular stimuli requires IKKα and IKKβ kinase activity . How IKKα and IKKβ are activated by various upstream signaling molecules is not fully understood . We previously showed that protein kinase C-associated kinase ( P03952 , also known as P57078 /RIP4 ) , which belongs to the receptor-interacting protein ( RIP ) kinase family , mediates the B cell activating factor of the P01375 family ( Q9Y275 ) -induced NF-κB activation in diffuse large B cell lymphoma ( DLBCL ) cell lines . Here we have investigated the mechanism underlying NF-κB activation regulated by P03952 . Our results suggest that P03952 can activate both the classical and the alternative NF-κB activation pathways . P03952 associates with IKKα and IKKβ in mammalian cells and induces activation of both IKKα and IKKβ via phosphorylation of their serine residues 176/180 and 177/181 , respectively . Unlike other members of the RIP family that activate NF-κB through a kinase-independent pathway , P03952 appears to activate IKK and NF-κB mainly in a kinase-dependent manner . Suppression of P03952 expression by RNA interference inhibits phosphorylation of IKKα and IKKβ as well as activation of NF-κB in human cancer cell lines . Thus , P03952 regulates NF-κB activation by modulating activation of IKKα and IKKβ in mammalian cells . We propose that P03952 may provide a critical link between IKK activation and various upstream signaling cascades , and may represent a potential target for inhibiting abnormal NF-κB activation in human cancers . Homocysteine increases methionine synthase mRNA level in Caco-2 cells . BACKGROUND : Q99707 ( Q99707 ) synthesizes methionine from homocysteine , using cobalamin as a cofactor and 5-methyltetrahydrofolate as a cosubstrate . AIM : To determine the influence of homocystine ( Hcy , dimer of homocysteine ) in the presence of either cobalamin or methionine on the transcription and the activity of methionine synthase in Caco-2 , a human adenocarcinoma cell line . METHODS : Q99707 activity and quantification of its mRNA by real-time RT-PCR were determined in cells cultivated under four differents conditions : Hcy with cobalamin ( Hcy+ Cbl+ ) , Hcy with methionine ( Hcy+ DB00134 + ) , methionine with Cbl ( DB00134 + Cbl+ ) and methionine only ( DB00134 + ) . RESULTS : Activity ( nmol/h/mg protein ) was maximal in cells cultivated in Hcy+Cbl+ ( 2.45 +/- 0.35 ) , compared to cells cultivated in Hcy+ DB00134 + ( 0.18 +/- 0.01 , p < 0.001 ) , in DB00134 + Cbl+ ( 1.60 +/- 0.06 , p < 0.05 ) , and in DB00134 + ( 0.40 +/- 0.05 , p < 0.001 ) , suggesting an adaptation of the cells to requirement in synthesized methionine . The mRNA level of Q99707 in Hcy+ Cbl+ and Hcy+ DB00134 + ( 2.82 +/-0.49 and 3.33 +/- 0.48 AU , respectively ) was about 2.5 / 3.0-fold higher than that in DB00134 + Cbl+ and in DB00134 + ( 1.00 +/-0.13 and 1.20 +/-0.20 AU , respectively , p < 0.001 ) . CONCLUSION : Q99707 expression of Caco-2 cell is under a transcriptional control influenced by Hcy . A comparison of the upper gastrointestinal mucosal effects of valdecoxib , naproxen and placebo in healthy elderly subjects . BACKGROUND : In long-term outcomes studies , cyclooxygenase P35354 specific inhibitors spare P23219 at supratherapeutic doses and therefore demonstrate improved gastrointestinal safety over nonspecific nonsteroidal anti-inflammatory drugs ( NSAIDs ) . However , in clinical practice , anti-inflammatory drugs are often used for short-term treatment of pain . AIM : To compare the short-term upper gastrointestinal mucosal effects of naproxen with the new P35354 specific inhibitor , valdecoxib , or placebo , in elderly subjects . METHODS : In this multicentre , double-blind , randomized , study , elderly subjects ( 65-76 years old ) , with a normal baseline esophagogastroduodenoscopy ( EGD ) , received oral valdecoxib ( a supratherapeutic 40 mg b.d. dosage , n = 62 ) , naproxen ( 500 mg b.d. , n = 62 ) , or placebo ( n = 62 ) for 6.5 days . Upper gastrointestinal mucosal injury was evaluated post-treatment by EGD ( day 7 ) . RESULTS : Subjects receiving naproxen ( 11/60 , 18 % ) had significantly more gastroduodenal ulcers post-treatment than those receiving placebo ( 2/61 , 3 % ; P < 0.01 ) or valdecoxib ( 0/60 , 0 % ; P < 0.001 ) . A similar significant finding was observed for gastric ulcer rates . All treatments had similar adverse event rates and clinical laboratory findings . CONCLUSIONS : DB00580 , even at supratherapeutic doses , was associated with an ulcer rate significantly lower than naproxen but similar to placebo in healthy elderly subjects , despite the short duration of therapy ( 6.5 days ) . Naproxen and valdecoxib were as well tolerated as placebo . MR properties of excised neural tissue following experimentally induced inflammation . Changes in the MR parameters of inflamed neural tissue were measured in vitro . P01375 -alpha ( P01375 ) was injected into rat sciatic nerves to induce inflammation with negligible axonal loss and demyelination . The MR parameters , such as T1/ P24752 relaxation and magnetization transfer ( MT ) , were measured 2 days after P01375 injection and were found to be substantially different from those of normal nerves . The average T1/ P24752 relaxation times increased , whereas the MT ratio ( Q99707 ) and the quantitative MT parameter M0B ( which describes the semisolid pool of protons ) decreased . The MR parameters correlated very well with the extracellular volume fraction ( EM ) of neural tissue evaluated by quantitative histopathology . The multicomponent P24752 relaxation was shown to provide the best quantitative assessment of changes in neural tissue microstructure , and allowed us to distinguish between the processes of inflammation and demyelination . In comparison , the MT measurements were less successful due to competing contributions of demyelination and pH-sensitive changes in the MT effect . Regulation of Con A-dependent cytokine production from P01730 + and CD8+ T lymphocytes by autosecretion of histamine . OBJECTIVES : Previously we have shown that both P01730 + T cells and CD8+ T cells produce histamine when activated with Con A . The aim of this study was to examine whether cytokine production by these cells is regulated by autosecretion of histamine . MATERIALS : P01730 + and CD8+ T cells were separated from spleen cells of C57BL/6 mice and mice lacking the H1 receptor ( P35367 ) or P25021 , using anti- P01730 +- and anti-CD8+-coupled magnetic beads , respectively . RESULTS : Depletion of the P35367 resulted in decreases in the release of P60568 and P22301 from both P01730 + and CD8+ cells and increases in the release of P05112 from P01730 + T cells and P01579 from CD8+ cells . Mice lacking the P25021 showed up-regulation of P01579 secretion from CD8+ cells and of P05112 from P01730 + and CD8+ T cells . Release of P60568 and P22301 from P01730 + as well as CD8+ cells was down-regulated in these mice . Both P01730 + and CD8+ T cell fractions synthesized histamine , which was enhanced in the P35367 -deficient CD8+ T cells . Treatment of the cells with alpha-fluoromethyl-histidine , a specific inhibitor of HDC , or histaminase increased P01579 from CD8+ cells , whereas it had no appreciable effect on P05112 secretion from P01730 + cells . CONCLUSION : These results suggest that cytokine production by P01730 + and CD8+ T lymphocytes is regulated by autosecretion of histamine . DB05311 ( DX-88 ) , a plasma kallikrein inhibitor for the treatment of hereditary angioedema and the prevention of blood loss in on-pump cardiothoracic surgery . BACKGROUND : P03952 plays a major role in the contact ( kallikrein-kinin ) cascade producing bradykinin . Bradykinin is a vasodilator , which increases vascular permeability , activates inflammation and produces pain . P03952 is also crosslinked to the coagulation system and the complement cascade . OBJECTIVE : DB05311 ( DX-88 ) is a potent and specific inhibitor of plasma kallikrein . DB05311 is a recombinantly produced and engineered small protein based on the first Kunitz domain of human tissue factor pathway inhibitor . It was identified through phage display technology . METHODS : The search terms ' ecallantide ' , ' DX-88 ' and ' hereditary angioedema ' were entered into Pubmed/Medline , ClinicalTrials and Google . RESULTS/CONCLUSION : At present , the drug is being studied for two major indications . First , the results for the treatment of hereditary angioedema are promising . Second , a prospective randomised multi-centre trial for the reduction of blood loss during on-pump cardiothoracic surgery will be terminated in October 2008 . An oligonucleotide blocks interferon-gamma signal transduction . Interferon ( IFN ) -gamma is an important mediator of transplant graft rejection . It induces endothelial cell expression of HLA-DR and intercellular adhesion molecule-1 , which render transplant grafts more susceptible to rejection by the host . Oligonucleotide 5'-GGG GTT P19440 TGT GTT GGG TGT TGT GT- Q96MN2 ( oligo I ) blocks multiple P01579 effects in human K562 cell cultures . A systematic approach revealed that oligo I has a novel , and potentially important , mode of action -- it blocks the binding of P01579 to its receptor , thus preventing activation of the P01579 signal transduction pathway . The results are consistent with an aptamer mechanism of action , because oligo I exerts its inhibitory effects by interacting with protein , not intracellular nucleic acid targets , such as mRNA or genomic DNA . P25021 mediated relaxation of buffalo ( Bubalus bubalus ) ureter . On the buffalo ureter , histamine did not elicit any direct effect . However , it caused concentration-dependent relaxation of the tissues precontracted by carbachol , phenylephrine , norepinephrine , KCI or BaCl2 and also inhibited the contractile effect of carbachol . DB08805 selectively antagonised the relaxation and inhibition of contractile response but mepyramine did not show this effect . Isoprenaline , dobutamine , salbutamol , verapamil and papaverine neither produced any direct effect nor relaxed the carbachol-contracted tissues ; norepinephrine and epinephrine had contractile effects . Hence , the histamine-induced relaxation was mediated through the activation of H2 receptors and not through adrenergic mechanisms or blockade of Ca(2+)-channels or inhibition of cyclic nucleotide phosphodiesterase . DB01296 improves cardiac function following trauma-hemorrhage by increased protein O-GlcNAcylation and attenuation of NF-{kappa}B signaling . We have previously demonstrated that in a rat model of trauma-hemorrhage ( T-H ) , glucosamine administration during resuscitation improved cardiac function , reduced circulating levels of inflammatory cytokines , and increased tissue levels of O-linked N-acetylglucosamine ( O-GlcNAc ) on proteins . The mechanism(s) by which glucosamine mediated its protective effect were not determined ; therefore , the goal of this study was to test the hypothesis that glucosamine treatment attenuated the activation of the nuclear factor-kappaB ( NF-kappaB ) signaling pathway in the heart via an increase in protein O-GlcNAc levels . Fasted male rats were subjected to T-H by bleeding to a mean arterial blood pressure of 40 mmHg for 90 min followed by resuscitation . DB01296 treatment during resuscitation significantly attenuated the T-H-induced increase in cardiac levels of P01375 and P05231 mRNA , IkappaB-alpha phosphorylation , NF-kappaB , NF-kappaB DNA binding activity , P05362 , and P05164 activity . LPS ( 2 microg/ml ) increased the levels of IkappaB-alpha phosphorylation , P01375 , P05362 , and NF-kappaB in primary cultured cardiomyocytes , which was significantly attenuated by glucosamine treatment and overexpression of O-GlcNAc transferase ; both interventions also significantly increased O-GlcNAc levels . In contrast , the transfection of neonatal rat ventricular myocytes with O15294 small-interfering RNA decreased O-GlcNAc transferase and O-GlcNAc levels and enhanced the LPS-induced increase in IkappaB-alpha phosphorylation . DB01296 treatment of macrophage cell line RAW 264.7 also increased O-GlcNAc levels and attenuated the LPS-induced activation of NF-kappaB . These results demonstrate that the modulation of O-GlcNAc levels alters the response of cardiomyocytes to the activation of the NF-kappaB pathway , which may contribute to the glucosamine-mediated improvement in cardiac function following hemorrhagic shock . The glial cell modulator and phosphodiesterase inhibitor , DB05066 ( ibudilast ) , attenuates prime- and stress-induced methamphetamine relapse . Stress and renewed contact with drug ( a " slip " ) have been linked to persisting relapse of methamphetamine abuse . Human brain microglial activation has been linked with methamphetamine abuse , and inhibitors of glial cell activation , certain phosphodiesterase ( PDE ) inhibitors , and glial cell derived neurotrophic factor ( P39905 ) have been reported to modulate drug abuse effects . Our objective was to determine whether the glial cell attenuator , 3-isobutyryl-2-isopropylpyrazolo-[1,5-a]pyridine ( DB05066 , ibudilast ) , a non-selective PDE inhibitor and promoter of P39905 , could reduce stress- and methamphetamine prime-induced reinstatement of methamphetamine-seeking behavior . Male Long-Evans hooded rats were trained to lever press reinforced with 0.1 mg/kg i.v. methamphetamine infusion according to fixed-ratio 1 ( FR1 ) reinforcement schedules during daily , 2-hour experimental sessions . After performance had stabilized , lever pressing was extinguished for 12 consecutive sessions and doses of 0 ( vehicle ) , 2.5 and 7.5 mg/kg DB05066 were then administered intraperitoneally b.i.d. on the last 2 days of extinction and then once on the testday to separate groups of 12 rats . During testing , the rats were given 15 min of intermittent footshock or a 1 mg/kg i.p. methamphetamine prime followed by a 2-hour reinstatement test session . DB05066 significantly reduced response levels of footshock-induced ( 2.5 and 7.5 mg/kg ) and prime-induced ( 7.5 mg/kg ) reinstatement of extinguished methamphetamine-maintained responding . DB05066 has properties consistent with the ability to attenuate relapse precipitated by stress and methamphetamine " slips " during abstinence . These results thus reinforce interest in atypical neurobiological mechanisms which could be exploited for developing novel medications for treating drug abuse disorders . Nonsteroidal anti-inflammatory drugs ( NSAID ) sparing effects of glucosamine hydrochloride through N-glycosylation inhibition ; strategy to rescue stomach from NSAID damage . Gastrointestinal or cardiovascular complications limit nonsteroidal anti-inflammatory drugs ( NSAID ) prescription . DB01296 hydrochloride ( GS-HCl ) alternatively chosen , but debates still exist in its clinical efficiency . P35354 instability through inhibiting P35354 N-glycosylation of GS-HCl raised the possibility of NSAID sparing effect . Study was done to determine whether combination treatment of glucosamine and NSAID contributes to gastric safety through NSAID sparing effect . IEC-6 cells were stimulated with P01375 -α and compared the expressions of inflammatory mediators after indomethacin alone or combination of indomethacin and GS-HCl by Western blotting and RT-PCR . C57BL/6 mice injected with type II collagen to induce arthritis were treated with indomethacin alone or combination of reduced dose of indomethacin and GS-HCl after 3 weeks . P01375 -α increased the expression of P35354 , P35228 and inflammatory cytokines , but GS-HCl significantly attenuated P01375 -α-induced P35354 expression . Decreased P35354 after GS-HCl was caused by N-glycosylation inhibition as much as tunicamycin . Combination of reduced dose of indomethacin and GS-HCl significantly reduced the expressions of P05362 , P19320 , P10145 , IL-1β , P08253 , P09237 , P14780 , and P24347 mRNA as well as NF-κB activation better than high dose indomethacin alone . These NSAID sparing effect of GS-HCl was further proven in collagen-induced arthritis model . Combination of GS-HCl and 2.5 mg/kg indomethacin showed significant protection from gastric damages as well as efficacious anti-arthritic effect . Taken together , P35354 N-glycosylation inhibition by GS-HCl led to indomethacin sparing effects , based on which combination of GS-HCl and reduced dose of NSAID can provide the strategy to secure stomach from NSAID-induced gastric damage as well as excellent anti-arthritic effects .

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interacts_with DB01197?

multiple_choice

DB04873 ( SB 207266 ) , a selective Q13639 receptor antagonist , reduces serotonin potentiation of neurally-mediated contractile responses of human detrusor muscle . The aim of this study is to evaluate the potency of piboserod ( SB 207266 ) , a selective 5-HT(4) receptor antagonist , at inhibiting the 5-HT(4)-mediated potentiating effect of serotonin ( 5-HT ) on the neurally-mediated contractile responses of human detrusor strips to electrical field stimulations ( O43281 ) . Strips of human detrusor muscle were mounted in Krebs-HEPES buffer under a resting tension of 500 mg and O43281 ( 20 Hz , 1 ms duration at 300 mA for 5 s ) was applied continuously at 1 min intervals . After stabilization of the O43281 -induced contractions , concentration-response curves to 5-HT ( 0.1 nM-100 microM ) were constructed in the absence or presence of 1 or 100 nM of piboserod . The experiments were performed in the presence of methysergide ( 1 microM ) and ondansetron ( 3 microM ) to block 5HT(1)/5HT(2) and 5-HT(3) receptors , respectively . 5-HT potentiated the contractile responses to O43281 of human bladder strips in a concentration-dependent manner , with a maximum mean of 60.0+/-19.9 % of the basal O43281 -evoked contractions . DB04873 did not modify the basal contractions but concentration-dependently antagonized the ability of 5-HT to enhance bladder strip contractions to O43281 . In presence of 1 and 100 nM of piboserod , the maximal 5-HT-induced potentiations were reduced to 45.0+/-7.9 and 38.7+/-8.7 % , respectively . A mean apparent antagonist dissociation constant value ( K(B) ) of 0.56+/-0.09 nM was determined . These data show the ability of piboserod to antagonize with high potency the enhancing properties of 5-HT on neurally-mediated contractions of isolated human bladder strips . Therefore , the 5-HT(4) receptor might represent an attractive pharmacological target for the treatment of overactive bladder . [ The effect of bunazosin vs captopril on hemodynamic and neurohumoral parameters in patients with congestive heart failure ] . The hemodynamic parameters ( right atrial pressure , mean pulmonary artery pressure , pulmonary capillary wedge pressure , cardiac index , heart rate , blood pressure ) and neurohumoral responses ( alpha- P01160 , plasma renin activity , aldosterone , angiotensin II ) of DB01197 , oral P12821 inhibitor , and Bunazosin , oral alpha 1-blocker , were investigated in 28 patients with congestive heart failure at rest and after exercise . These data were analysed in both acute and chronic phases . 1 ) Acute effect . DB01197 produced significant improvement of neurohumoral factors at rest and also after exercise . Bunazosin reduced alpha- P01160 , but other neurohumoral factors did not change . Bunazosin produced significant hemodynamic improvement both at rest and after exercise . 2 ) Chronic effect . DB01197 produced significant hemodynamic improvement both at rest and after exercise . Improvement of neurohumoral factors in acute phase was also preserved at chronic phase . On Bunazosin , improvement of hemodynamics at acute phase was also preserved at chronic phase without deterioration of neurohumoral factors . Genetic variants and multiple myeloma risk : IMMEnSE validation of the best reported associations -- an extensive replication of the associations from the candidate gene era . BACKGROUND : Genetic background plays a role in multiple myeloma susceptibility . Several single-nucleotide polymorphisms ( SNP ) associated with genetic susceptibility to multiple myeloma were identified in the last years , but only a few of them were validated in independent studies . METHODS : With the aim to conclusively validate the strongest associations so far reported , we selected the polymorphisms rs2227667 ( P05121 ) , rs17501108 ( P14210 ) , rs3136685 ( P32248 ) , rs16944 ( P01584 ) , rs12147254 ( Q13114 ) , rs1805087 ( Q99707 ) , rs1800629 ( P01375 -α ) , rs7516435 ( P55211 ) , rs1042265 ( Q07812 ) , rs2234922 ( mEH ) , and rs1801133 ( P42898 ) . We genotyped them in 1,498 multiple myeloma cases and 1,934 controls ascertained in the context of the International Multiple Myeloma rESEarch ( IMMEnSE ) consortium , and meta-analyzed our results with previously published ones . RESULTS : None of the selected SNPs were significantly associated with multiple myeloma risk ( P value range , 0.055-0.981 ) , possibly with the exception of the SNP rs2227667 ( P05121 ) in women . CONCLUSIONS : We can exclude that the selected polymorphisms are major multiple myeloma risk factors . Q9P2X3 : Independent validation studies are crucial to identify true genetic risk factors . Our large-scale study clarifies the role of previously published polymorphisms in multiple myeloma risk . A P43220 agonist liraglutide inhibits endothelial cell dysfunction and vascular adhesion molecule expression in an ApoE-/- mouse model . The glucagon like peptide-1 receptor ( P43220 ) agonist liraglutide attenuates induction of plasminogen activator inhibitor type-1 ( P05121 ) and vascular adhesion molecule ( VAM ) expression in human vascular endothelial cells ( hVECs ) in vitro and may afford protection against endothelial cell dysfunction ( O95905 ) , an early abnormality in diabetic vascular disease . Our study aimed to establish the dependence of the in vitro effects of liraglutide on the P43220 and characterise its in vivo effects in a mouse model of O95905 . In vitro studies utilised the human vascular endothelial cell line P10144 - Q8IWL8 and enzyme-linked immunosorbent assays ( ELISA ) for determination of P05121 and VAM expression . In vivo studies of vascular reactivity and immunohistochemical analysis were performed in the ApoE(-/-) mouse model . In vitro studies demonstrated P43220 -dependent liraglutide-mediated inhibition of stimulated P05121 and VAM expression . In vivo studies demonstrated significant improvement in endothelial function in liraglutide treated mice , a P43220 dependent effect . DB06655 treatment also increased endothelial nitric oxide synthase ( P29474 ) and reduced intercellular adhesion molecule-1 ( P05362 ) expression in aortic endothelium , an effect again dependent on the P43220 . Together these studies identify in vivo protection , by the P43220 agonist liraglutide , against O95905 and provide a potential molecular mechanism responsible for these effects . Expression of Q07812 in cell nucleus after experimentally induced apoptosis revealed by immunogold and embedment-free electron microscopy . The unique combination of immunocytochemistry with embedment-free electron microscopy was applied for precise and specific localisation of Q07812 in the human colon adenocarcinoma COLO 205 cell line stimulated to undergo apoptosis by camptothecin ( P11387 inhibitor ) . DB04690 -induced apoptosis was associated with redistribution of Q07812 from cytosol to organelle membranes : mitochondria , Golgi apparatus , endoplasmic reticulum and via nuclear envelope pores to the nucleus , occurring within 60-180 min of cell exposure to the drug . An increase in Q07812 immunoreactivity on fine filaments and the lamina-pore complex of the nuclear matrix was also observed . The increase in Q07812 expression in the nuclear area of camptothecin-treated COLO 205 cells was confirmed by quantitative analysis using laser scanning cytometry . The subcellular translocations of Q07812 preceded the appearance of any morphological symptoms of apoptosis . Angiotensin-converting-enzyme inhibitors suppress synthesis of tumour necrosis factor and interleukin 1 by human peripheral blood mononuclear cells . Administration of angiotensin-converting-enzyme ( P12821 ) inhibitors reduce vascular proliferation following endothelial injury as well as progression of renal disease in various animal models . These effects might be due to interference with cytokines such as interleukin 1 ( IL-1 ) or tumour necrosis factor alpha ( P01375 ) since they have been implicated in regulating the effects of vascular cell growth factors such as fibroblast- and platelet-derived growth factors . We investigated the in vitro synthesis of IL-1 and P01375 from human peripheral blood mononuclear cells ( PBMC ) in the presence of various P12821 -inhibitors . DB01197 dose-dependently suppressed the P01584 -induced synthesis of P01375 by 74 % ( P < 0.01 ) and the P01584 -induced synthesis of P01583 by 60 % ( P < 0.01 ) . Cytokine synthesis induced by lipopolysaccharide was less affected . At concentrations suppressing P01375 and IL-1 , captopril did not reduce the synthesis of complement P01024 in the same cells . Enalapril and cilazapril also suppressed cytokine-induced cytokine synthesis . Ramipril , lisinopril , perindopril and spirapril had no significant effect on P01375 synthesis suggesting that the effect was not related specifically to the inhibition of P12821 . Accumulation of mRNA for IL-1 and P01375 were not affected by captopril , suggesting a posttranscriptional effect . We conclude that certain P12821 -inhibitors suppress IL-1 and P01375 synthesis at a posttranscriptional level and might therefore influence cytokine-mediated cell growth . [ DB09043 ( Eperzan ) : a new once-weekly agonist of glucagon-like peptide-1 receptors ] . DB09043 ( Eperzan ) is a new once-weekly agonist of Glucagon-Like Peptide-1 ( P0C6A0 ) receptors that is indicated in the treatment of type 2 diabetes . Two doses are available , 30 mg and 50 mg , to be injected subcutaneously once a week . It has been extensively evaluated in the HARMONY programme of eight large randomised controlled trials that were performed at different stages of type 2 diabetes , in comparison with placebo or an active comparator . The endocrine and metabolic effects of albiglutide are similar to those of other P43220 agonists : stimulation of insulin secretion ( incretin effect ) and inhibition of glucagon secretion , both in a glucose-dependent manner , retardation of gastric emptying and increase of satiety . These effects lead to a reduction in glycated haemoglobin ( HbA(1c) ) levels , combined with a weight reduction . The overall tolerance profile is good . DB09043 is currently reimbursed in Belgium after failure ( HbA(1c) > 7.5 % ) of and in combination with a dual therapy with metformin and a sulfonylurea as well as in combination with a basal insulin ( with or without oral antidiabetic drugs ) . To avoid hypoglycaemia , a reduction in the dose of sulfonylurea or insulin may be recommended . A once-weekly administration should increase patient 's acceptance of injectable therapy and improve compliance . DB01954 attenuates bile duct ligation-induced liver injury in rats : a potential pathogenic role of DB05876 . Anti-inflammatory and antifibrotic effects of the broad spectrum phosphodiesterase ( PDE ) inhibitor pentoxifylline have suggested an important role for cyclic nucleotides in the pathogenesis of hepatic fibrosis ; however , studies examining the role of specific PDEs are lacking . Endotoxemia and O00206 ( O00206 ) -mediated inflammatory and profibrotic signaling play a major role in the development of hepatic fibrosis . Because DB02527 -specific DB05876 critically regulates lipopolysaccharide ( LPS ) - O00206 -induced inflammatory cytokine expression , its pathogenic role in bile duct ligation-induced hepatic injury and fibrogenesis in Sprague-Dawley rats was examined . Initiation of cholestatic liver injury and fibrosis was accompanied by a significant induction of P27815 , B , and D expression and activity . Treatment with the DB05876 -specific inhibitor rolipram significantly decreased liver DB05876 activity , hepatic inflammatory and profibrotic cytokine expression , injury , and fibrosis . At the cellular level , in relevance to endotoxemia and inflammatory cytokine production , Q07343 was observed to play a major regulatory role in the LPS-inducible tumor necrosis factor ( P01375 ) production by isolated Kupffer cells . Moreover , DB05876 expression was also involved in the in vitro activation and transdifferentiation of isolated hepatic stellate cells ( HSCs ) . Particularly , P27815 , B , and D upregulation preceded induction of the P19526 activation marker α-smooth muscle actin ( α-SMA ) . In vitro treatment of HSCs with rolipram effectively attenuated α-SMA , collagen expression , and accompanying morphologic changes . Overall , these data strongly suggest that upregulation of DB05876 expression during cholestatic liver injury plays a potential pathogenic role in the development of inflammation , injury , and fibrosis . Effects of a Topical Angiotensin-Converting Enzyme Inhibitor and a Selective P35354 Inhibitor on the Prevention of Hypertrophic Scarring in the Skin of a Rabbit Ear .  P12821 ( P12821 ) inhibitors have been reported to inhibit fibrogenesis , and cyclooxygenase-2 ( P35354 ) inhibitors , to reduce scarring by reducing the initial inflammation . The authors reasoned that the topical application of these 2 agents may have a complementary effect on scar reduction . METHODS : DB01197 ( P12821 inhibitor ) , celecoxib ( P35354 inhibitor ) , or a combination of captopril and celecoxib were topically applied to a skin wound in a rabbit ear , and investigated for the effects on scar formation . RESULTS : The level of scar elevation decreased in the captopril group and the level of infiltration of inflammatory cells decreased in the celecoxib group . In the group where a combination of the 2 drugs was used , the level of scar elevation decreased the most , and collagen deposition and organization returned to normal most rapidly . Celecoxib was found to inhibit the initial inflammation in the ear wound of the rabbit , and captopril inhibited scar elevation . CONCLUSION : Clinical application of these drugs will require further studies with regard to adverse events and their absorptivity as topical agents . However , these findings suggest that the combined topical administration of an P12821 inhibitor and P35354 inhibitor to a skin wound could be an effective treatment for the prevention of hypertrophic scarring.  . Chronic atrial fibrillation alters the functional properties of If in the human atrium . INTRODUCTION : Despite the evidence that the hyperpolarization-activated current ( If ) is highly modulated in human cardiomyopathies , no definite data exist in chronic atrial fibrillation ( cAF ) . We investigated the expression , function , and modulation of If in human cAF . METHODS AND RESULTS : Right atrial samples were obtained from sinus rhythm ( SR , n = 49 ) or cAF ( duration > 1 year , n = 31 ) patients undergoing corrective cardiac surgery . Among f-channel isoforms expressed in the human atrium ( O60741 , 2 and 4 ) , Q9Y3Q4 mRNA levels measured by RT-PCR were significantly reduced . However , protein expression was preserved in cAF compared to SR ( +85 % for Q9Y3Q4 ) ; concurrently , miR-1 expression was significantly reduced . In patch-clamped atrial myocytes , current-specific conductance ( gf ) was significantly increased in cAF at voltages around the threshold for If activation ( -60 to -80 mV ) ; accordingly , a 10-mV rightward shift of the activation curve occurred ( P < 0.01 ) . β-Adrenergic and Q13639 receptor stimulation exerted similar effects on If in cAF and SR cells , while the P01160 -mediated effect was significantly reduced ( P < 0.02 ) , suggesting downregulation of natriuretic peptide signaling . CONCLUSIONS : In human cAF modifications in transcriptional and posttranscriptional mechanisms of HCN channels occur , associated with a slight yet significant gain-of-function of If , which may contribute to enhanced atrial ectopy . Secretome profiling reveals the signaling molecules of apoptotic HCT116 cells induced by the dietary polyacetylene gymnasterkoreayne B . Dietary polyacetylenes from various foods have been receiving attention as promising cancer chemopreventive agents . However , until now , the detailed molecular mechanism and the regulatory proteins underlying these effects have not been elucidated . We investigated the effects of gymnasterkoreayne B ( GKB ) , a model dietary polyacetylene from wild vegetables , on the programmed cell death of HCT116 human colorectal cancer cells . GKB inhibited HCT116 cell proliferation by inducing apoptotic cell death . GKB treatment resulted in ROS accumulation , leading to the activation of both intrinsic and extrinsic apoptotic pathway . We also found that P02751 , P01137 , P05067 , P05121 , P10809 , P00441 , P10599 , and O43707 may act as secretory signaling molecules during GKB-induced apoptotic cell death using LC-MS/MS identification followed by spectrum counting , statistical calculation , and gene ontology analysis . The secretory proteins suggested in this study may be promising candidates involved in apoptotic cell death of cancer cells induced by GKB that warrant further functional study . Clinical and pathologic perspectives on aspirin sensitivity and asthma . DB00945 and other nonsteroidal anti-inflammatory drugs that inhibit P23219 induce unique nonallergic reactions , consisting of attacks of rhinitis and asthma . These hypersensitivity reactions occur in a subset of asthmatic subjects , thus identifying them as having this exclusive clinical presentation . We refer to these patients as having aspirin-exacerbated respiratory disease , a disease process that produces devastating eosinophilic inflammation of both the upper and lower respiratory tracts . This review focuses on a description of patients with aspirin-exacerbated respiratory disease , methods available to diagnose their condition , the unique ability of all nonsteroidal anti-inflammatory drugs that inhibit P23219 to cross-react with aspirin , an update on pathogenesis , and current thoughts about treatment . Q92847 agonist ( DB05657 ) accelerates gastric emptying in adults with diabetes and symptomatic gastroparesis . BACKGROUND : DB05657 is a synthetic , selective ghrelin agonist in development for gastroparesis . AIM : To assess safety and effects of DB05657 in diabetes patients with symptomatic gastroparesis . METHODS : Adults with type 1 or type 2 diabetes mellitus received placebo and DB05657 ( 80 , 160 , 320 or 600 microg/kg ) infusions in a cross-over manner following a radiolabelled meal . Blood glucose levels were stabilized using a hyperinsulinemic-euglycemic clamp . Primary endpoints were gastric half emptying and latency times . Secondary measures included assessment of gastroparesis symptoms and endocrine responses . RESULTS : Ten patients with type 1 ( n = 7 ) or 2 ( n = 3 ) diabetes , moderate-to-severe gastroparesis symptoms and > or =29 % retention 4 h after a radiolabelled solid meal were enrolled . DB05657 produced significant reductions in solid meal half-emptying ( 20 % , P = 0.043 ) and latency ( 34 % , P = 0.037 ) times vs. placebo . Reductions in overall postmeal symptom intensity ( 24 % ) and postprandial fullness ( 37 % ) following DB05657 infusion were not statistically significant . Most adverse events were mild and self-limiting and there were no identifiable differences in numbers or types of adverse events between DB05657 and placebo . CONCLUSIONS : This proof-of-concept study demonstrates that the ghrelin agonist DB05657 is well-tolerated in diabetes patients with moderate-to-severe chronic gastroparesis and shows statistically significant improvements in gastric emptying . The use of a cyclooxygenase-2 inhibitor ( DB06802 ) in an ocular and metastatic animal model of uveal melanoma . The expression of cyclooxygenase-2 ( P35354 ) has been reported as an indicator of poor prognosis in a wide variety of human tumors , including colon , breast and uveal melanoma ( UM ) . P35354 inhibitors have shown promise in controlling the malignancy of several types of tumors . Previous studies have demonstrated the efficacy of a P35354 inhibitor on the proliferation rates of human UM cells . The goal of this experiment was to investigate the efficiency of DB06802 , a topically administered P35354 inhibitor , in a rabbit model of UM . The animals were divided into two groups of 14 animals for the duration of the 12-week experiment . One animal per group was killed each week to evaluate disease progression and for histopathological studies . The experimental group received drops containing 0.3 % DB06802 solution . Intraocular tumor growth was evaluated weekly by fundoscopic examination and each animal was weighed prior to examination . Blood samples were taken weekly from all rabbits to detect circulating malignant cells ( CMCs ) throughout the experiment . After the second week of inoculation , the experimental group weighed significantly more than the control group . The control group developed more intraocular tumors and presented with metastases and higher detectable levels of CMCs before the treated group . These results indicate that the topical administration of a P35354 inhibitor delayed the progression of this malignancy in our animal model . A clinical trail using an anti- P35354 inhibitor for patients with UM should be considered . A pleiotropic antiatherogenic action of ibuprofen . Ibuprofen is a cyclooxygenase ( P23219 and P35354 ) inhibitor known to reduce the production of prostaglandins that play prominent role in inflammation . Other properties of the drug , aside from its anti-inflammatory effects , have been recently studied . In this paper we shall discuss several properties of ibuprofen that making the drug interesting for treatment of conditions associated with atherosclerosis . Ibuprofen exerts pleiotropic effects such as inhibition of adhesion and transendothelial migration of leukocytes , suppressing intracellular production of reactive oxygen species and oxidative modification of LDL . Interestingly , ibuprofen increased HDL cholesterol levels and reduced the level of triglicerides . Ibuprofen can also modulate efficiency of fibrynolisis by inhibiting production of plasminogen activator inhibitor ( P05121 ) . This properties of ibuprofen may be due to changing the activity of transcription factors . Ibuprofen inhibits the activation of NF-kB and activates PPARa and PPARg . DB00197 increases IL-1beta induced cyclooxygenase-2 and inducible nitric oxide synthase expression via enhanced phosphorylation of P25963 in vascular smooth muscle cells from Wistar-Kyoto rats and spontaneously hypertensive rats . P37231 ( PPARgamma ) agonists of the thiazolidinedione class are widely used for the treatment of type 2 diabetes subjects due to their ability to improve insulin resistance . DB00197 and ciglitazone belong to the PPARgamma agonists of thiazolidinediones . We report here that troglitazone but not ciglitazone increased IL-1beta induced cyclooxygenase-2 ( P35354 ) and inducible nitric oxide synthase ( P35228 ) expression in vascular smooth muscle cell ( VSMC ) from Wistar-Kyoto rats ( WKY ) and spontaneously hypertensive rats ( SHR ) . Potentiated expression of P35354 and P35228 by troglitazone was inhibited by MG-132 , a specific inhibitor of inhibitory factor kappaB ( IkappaB ) activation . DB00197 treatment of these cells also resulted in a dose-dependent increase in IL-1beta induced P25963 phosphorylation . These data suggest that troglitazone is capable of increasing IL-1beta induced P35354 and P35228 expression through an P25963 dependent mechanism in VSMC from WKY and SHR . DB01197 reduced plasminogen activator inhibitor activity in patients with acute myocardial infarction . Recent clinical trials have demonstrated that the administration of angiotensin-converting enzyme ( P12821 ) inhibitors to patients with myocardial infarction reduces the incidence of recurrent myocardial infarction . It has also been reported that an elevated level of plasminogen activator inhibitor ( P05121 ) appears to constitute a marker of the risk of recurrent coronary thrombosis . To determine whether the P12821 inhibitor captopril reduces plasma P05121 inhibitor activity , we measured changes in plasma P05121 activity ( IU/ml ) , tissue plasminogen activator ( t-PA ) antigen ( ng/ml ) , and serum P12821 activity ( IU/L ) in 14 survivors of myocardial infarction receiving captopril therapy ( 37.5 mg daily ) and compared them with the values in 15 placebo-treated patients chosen at random . Blood sampling was performed at 07.00 h . In the captopril-treated group , serum P12821 activity decreased significantly , from 14.0 +/- 0.8 to 11.5 +/- 1.2 IU/L 24 h after captopril therapy ( p < 0.01 ) , and those of P05121 activity and t-PA antigen also decreased significantly-from 11.9 +/- 2.8 to 5.5 +/- 2.2 IU/ml ( p < 0.02 ) and from 9.9 +/- 1.0 to 7.5 +/- 0.9 ng/ml ( p < 0.05 ) , respectively 48 h after captopril therapy . However , the levels of P12821 activity , P05121 activity , and t-PA antigen remained unchanged during the study period in the placebo group . Thus , our data indicate that the administration of captopril to patients with acute myocardial infarction may result in a reduced frequency of recurrent coronary thrombosis by increasing fibrinolytic capacity . DB05025 , a coinducer of heat shock proteins for the potential treatment of amyotrophic lateral sclerosis . Recent years have seen an explosion of research into increasingly prevalent neurodegenerative diseases . DB05025 ( BRX-220 ) , being developed by CytRx Corp , is an oral therapeutic candidate for the treatment of amyotrophic lateral sclerosis ( P35858 ) , the most common form of motor neuron disease . P35858 is a fatal , incurable disorder , which can present as sporadic ( 90 to 95 % of cases ) or familial ( 5 to 10 % of cases ) forms . The etiology of sporadic P35858 remains unknown and much of the understanding of P35858 pathogenesis has been derived through study of its familial forms ; in particular , through study of autosomal dominant mutations in the P00441 ( copper/zinc superoxide dismutase ) gene , which cause approximately 20 % of familial P35858 cases . Under conditions of excessive stress , arimoclomol induces amplification of the cytoprotective heat shock response in order to protect motor neurons from death . Comprehensive in vivo and in vitro studies demonstrated its effect in the prevention of neuronal loss and promotion of motor neuron survival , even after symptom onset . Clinical trials have reported good tolerability and safety . This paper discusses the rationale for arimoclomol use in P35858 , the preclinical and clinical evidence collected to date , the likelihood of its promising preclinical results translating to humans , and the relevance of this research for neurodegeneration as a whole . Ghrelin protects against renal damages induced by angiotensin-II via an antioxidative stress mechanism in mice . We explored the renal protective effects by a gut peptide , Ghrelin . Daily peritoneal injection with Ghrelin ameliorated renal damages in continuously angiotensin II ( AngII ) -infused C57BL/6 mice as assessed by urinary excretion of protein and renal tubular markers . AngII-induced increase in reactive oxygen species ( ROS ) levels and senescent changes were attenuated by Ghrelin . Ghrelin also inhibited AngII-induced upregulations of transforming growth factor-β ( TGF-β ) and plasminogen activator inhibitor-1 ( P05121 ) , ameliorating renal fibrotic changes . These effects were accompanied by concomitant increase in mitochondria uncoupling protein , P55851 as well as in a key regulator of mitochondria biosynthesis , PGC1α . In renal proximal cell line , HK-2 cells , Ghrelin reduced mitochondria membrane potential and mitochondria-derived ROS . The transfection of P55851 siRNA abolished the decrease in mitochondria-derived ROS by Ghrelin . Ghrelin ameliorated AngII-induced renal tubular cell senescent changes and AngII-induced TGF-β and P05121 expressions . Finally , Q92847 , growth hormone secretagogue receptor ( Q92847 ) -null mice exhibited an increase in tubular damages , renal ROS levels , renal senescent changes and fibrosis complicated with renal dysfunction . Q92847 -null mice harbored elongated mitochondria in the proximal tubules . In conclusion , Ghrelin suppressed AngII-induced renal damages through its P55851 dependent anti-oxidative stress effect and mitochondria maintenance . Ghrelin/ Q92847 pathway played an important role in the maintenance of ROS levels in the kidney .

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The somatostatin immunoregulatory circuit present at sites of chronic inflammation . Somatostatin is part of an immunoregulatory circuit that helps limit interferon-gamma ( IFNgamma ) production at sites of chronic inflammation . In murine schistosomiasis. parasite eggs induce focal , chronic granulomatous inflammation in the liver and intestines . These granulomas produce somatostatin 1-14 and express somatostatin receptor subtype number 2 ( P30874 ) , which is the exclusive somatostatin receptor present in this inflammation . Granuloma and splenic macrophages as well as macrophage cell lines make somatostatin . There appears to be no other inflammatory cell source of the peptide . Various inflammatory mediators induce this expression , whereas DB05875 inhibits somatostatin production . Somatostatin can suppress P01579 secretion from T cells via interaction with the P30874 receptor expressed on these cells . Other cells within the granuloma also display P30874 . The effect of somatostatin on these other cell types remains unknown . The thymus of normal mice has a complete somatostatin regulatory circuit . The thymic epithelial and dendritic cells make somatostatin . Like the granulomas of murine schistosomiasis , the thymus expresses only P30874 . Somatostatin likely has an important role in thymic T cell education and selection . Endothelin receptor antagonists as disease modifiers in systemic sclerosis . Systemic sclerosis ( SSc ) is a multisystem connective tissue disease of unknown etiology that is characterized by inflammation , vascular dysfunction and fibrosis of the skin and visceral organs . SSc is clinically diverse both in terms of the burden of skin and organ involvement and the rate of progression of the disease . Recent studies indicate that the endothelin system , especially ET-1 and the P25101 and ETB receptors may play a key role in the pathogenesis of SSc . A new class of drugs , endothelin receptor antagonists has been introduced for treatment of patients with pulmonary arterial hypertension ( PAH ) . DB00559 , a dual endothelin receptor antagonist as well as DB06268 and DB06403 , selective blockers of the P25101 receptor have proven effective in SSc-PAH . This effect may be mediated through both a vasodilatory and antifibrotic effect , thus making these agents attractive as potential disease modifying agents for SSc . P51955 mediates P00352 -dependent drug resistance in multiple myeloma . We reported previously that increased expression of aldehyde dehydrogenase 1 ( P00352 ) in multiple myeloma ( MM ) is a marker of tumor-initiating cells ( TICs ) that is further associated with chromosomal instability ( Q96GD0 ) . Here we demonstrate that member A1 of the P00352 family of proteins , P00352 , is most abundantly expressed in myeloma . Enforced expression of P00352 in myeloma cells led to increased clonogenicity , tumor formation in mice , and resistance to myeloma drugs in vitro and in vivo . The mechanism underlying these phenotypes included the P00352 -dependent activation of drug-efflux pump , P08183 , and survival proteins , AKT and P10415 . Over expression of P00352 in myeloma cells led to increased mRNA and protein levels of NIMA-related kinase 2 ( P51955 ) , whereas shRNA-mediated knock down of P51955 decreased drug efflux pump activity and drug resistance . The activation of P51955 in myeloma cells relied on the P00352 -dependent generation of the retinoid X receptor α ( RXRα ) ligand , 9-cis retinoic acid ( 9CRA ) - not the retinoic acid receptor α ( RARα ) ligand , all-trans retinoic acid ( DB00755 ) . These findings implicate the P00352 -RXRα- P51955 pathway in drug resistance and disease relapse in myeloma and suggest that specific inhibitors of P00352 are worthy of consideration for clinical development of new approaches to overcome drug resistance in myeloma . P01579 -dependent induction of human intercellular adhesion molecule-1 gene expression involves activation of a distinct P35610 protein complex . In response to interferon gamma ( IFNgamma ) , intercellular adhesion molecule-1 ( P05362 ) is expressed on human keratinocytes , a cell type that is critically involved in cutaneous inflammation . An P05362 5' regulatory region palindromic response element , pIgammaRE , has been shown to confer IFNgamma-dependent transcription enhancement . By electrophoretic mobility shift assays ( EMSA ) , pIgammaRE forms a distinct complex with proteins from IFNgamma-treated human keratinocytes , termed gamma response factor ( P01286 ) . Binding of P01286 is tyrosine phosphorylation-dependent , and mutations of pIgammaRE that disrupt the palindromic sequence or alter its spatial relationship abrogate P01286 binding . Supershift EMSAs using antibodies to characterized P35610 proteins suggest that P01286 contains a Stat1alpha-like protein ; however , non- P05362 IFNgamma-responsive elements ( REs ) known to bind Stat1alpha homodimers fail to compete for P01286 binding in EMSA , and pIgammaRE does not cross-compete with these REs that complex with homodimeric stat1alpha . The pIgammaRE x P01286 complex also displays a distinctly different electrophoretic mobility compared to that of IFNgammaREs complexed to homodimeric Stat1alpha . These findings indicate that a distinct complex containing a Stat1alpha-like protein mediates IFNgamma-induced P05362 gene transcription and identifies a subset of IFNgamma-responsive genes that appear to be regulated by this complex . Expression of retinoic acid receptor beta in dermatofibrosarcoma protuberans . BACKGROUND : P10826 ( RAR beta ) has been shown to act as a tumor suppressor in many solid human tumors . To investigate the putative role of RAR beta in dermatofibrosarcoma protuberans ( DFSP ) , we examined the expression of RAR beta in DFSPs and analyzed the correlation of expression patterns between RAR beta and cyclooxygenase ( P36551 ) -2 as well as clinicopathological variables . METHODS : Using tissue microarray and immunohistochemistry , we evaluated nuclear RAR beta staining and cytoplasm P35354 staining in 53 DFSPs . RESULTS : 48 DFSPs ( 90.58 % ) were immunopositive for RAR beta , while 32 DFSPs ( 60.38 % ) were immunopositive for P35354 . RAR beta staining was significantly inversely correlated with P35354 staining ( p < 0.001 ; r =-0.668 ) . CONCLUSIONS : Our data indicated that RAR beta expressed in DFSPs and correlated with P35354 expression . RAR beta may be a potential therapeutic target for unresectable DFSP cases . Expression of vitamin D3 receptor and retinoid receptors in human breast cancer : identification of potential heterodimeric receptors . DB00169 ( VD ) and all-trans-retinoic acid ( DB00755 ) have been postulated as a novel treatment option for breast carcinoma . Since the combined effects of retinoids and VD derivatives are attributed to heterodimeric interactions between members of the nuclear receptor family , the expression patterns of the heterodimers formed by vitamin D3 receptor ( P11473 ) and the retinoid receptors RARs ( P10276 , P10826 and P13631 ) and RXRs ( RXR-alpha , RXR-beta and RXR-gamma ) have been studied by immunohistochemistry in benign and malignant breast tissues . Present results revealed that immunoexpressions to all receptor types studied were higher in both in situ and infiltrative carcinomas than in benign breast diseases . In a variable number of cases of infiltrative carcinoma , immunostaining appeared in the nucleus , whereas in the other two disorders immunostaining was only cytoplasmic . The correlation established between P11473 and the different isoforms of retinoid receptors revealed that P11473 seems to select mainly P10276 to form heterodimers and to exert their properties as transcription factor . The results of this study suggest that this heterodimer plays a critical role in cancer malignancy , and its presence indicates those patient groups presenting a better response to adjuvant therapies based on the combination of vitamin D and DB00755 . Expression of the tyrosine phosphatase P12931 homology 2 domain-containing protein tyrosine phosphatase 1 determines T cell activation threshold and severity of experimental autoimmune encephalomyelitis . Experimental autoimmune encephalomyelitis ( EAE ) is a P01730 Th1-mediated inflammatory demyelinating disorder of the CNS and a well-established animal model for multiple sclerosis . Src homology 2 domain-containing protein tyrosine phosphatase 1 ( Q15466 -1 ) is a cytosolic tyrosine phosphatase that is involved in regulating the T cell activation cascade from signals initiated through the TCR . To study the role of Q15466 -1 in EAE pathogenesis , we immunized B10.PL mice heterozygous for deletion of the Q15466 -1 gene ( me(v+/-) ) and B10.PL wild-type mice with the immunodominant epitope of myelin basic protein ( MBP Ac1-11 ) . T cell proliferation and P01579 production were significantly increased in me(v+/-) mice after immunization with MBP Ac1-11 . The frequency of MBP Ac1-11-specific P01730 T cells , analyzed by staining with fluorescently labeled tetramers ( P11226 -11[4Y] : I-A(u) complexes ) , was increased in the draining lymph node cells of me(v+/-) mice compared with wild-type mice . In addition , me(v+/-) mice developed a more severe course of EAE with epitope spreading to proteolipid protein peptide 43-64 . Finally , expansion of MBP Ac1-11-specific T cells in response to Ag was enhanced in me(v+/-) T cells , particularly at lower Ag concentrations . These data demonstrate that the level of Q15466 -1 plays an important role in regulating the activation threshold of autoreactive T cells . The role of the adenosinergic system in lung fibrosis . DB00640 ( Q96SZ5 ) is a retaliatory metabolite that is expressed in conditions of injury or stress . During these conditions DB00171 is released at the extracellular level and is metabolized to adenosine . For this reason , adenosine is defined as a " danger signal " for cells and organs , in addition to its important role as homeostatic regulator . Its physiological functions are mediated through interaction with four specific transmembrane receptors called P30542 , P29274 , P29275 and P0DMS8 . In the lungs of mice and humans all four adenosine receptors are expressed with different roles , having pro- and anti-inflammatory roles , determining bronchoconstriction and regulating lung inflammation and airway remodeling . DB00640 receptors can also promote differentiation of lung fibroblasts into myofibroblasts , typical of the fibrotic event . This last function suggests a potential involvement of adenosine in the fibrotic lung disease processes , which are characterized by different degrees of inflammation and fibrosis . Idiopathic pulmonary fibrosis ( IPF ) is the pathology with the highest degree of fibrosis and is of unknown etiology and burdened by lack of effective treatments in humans . P22888 expression in leiomyomatosis peritonealis disseminata . BACKGROUND : Leiomyomatosis peritonealis disseminata has been attributed to estrogen stimulation and is seen only rarely in postmenopausal women . In such cases , pathogenesis is uncertain . CASE : Leiomyomatosis peritonealis disseminata tumors were resected from a postmenopausal woman . She was receiving tamoxifen therapy for breast cancer and had bilateral ovarian Brenner tumors . Estrogen and progesterone receptors were detected . Immunohistochemical analysis indicated that LH receptors were present . CONCLUSION : DB00044 receptors were identified in leiomyomatosis peritonealis disseminata in one woman . Levels of DB00094 and LH increase after menopause , and immunohistochemical analysis showed the presence of LH receptors , so gonadotropin rather than estrogen stimulation might have contributed to development of leiomyomatosis peritonealis disseminata in this uncommon case . Genome-wide association studies identify P30532 /3 and Q13639 in the development of airflow obstruction . RATIONALE : Genome-wide association studies ( GWAS ) have identified loci influencing lung function , but fewer genes influencing chronic obstructive pulmonary disease ( P48444 ) are known . OBJECTIVES : Perform meta-analyses of GWAS for airflow obstruction , a key pathophysiologic characteristic of P48444 assessed by spirometry , in population-based cohorts examining all participants , ever smokers , never smokers , asthma-free participants , and more severe cases . METHODS : Fifteen cohorts were studied for discovery ( 3,368 affected ; 29,507 unaffected ) , and a population-based family study and a meta-analysis of case-control studies were used for replication and regional follow-up ( 3,837 cases ; 4,479 control subjects ) . Airflow obstruction was defined as Q99581 (1) and its ratio to FVC ( Q99581 (1)/FVC ) both less than their respective lower limits of normal as determined by published reference equations . MEASUREMENTS AND MAIN RESULTS : The discovery meta-analyses identified one region on chromosome 15q25.1 meeting genome-wide significance in ever smokers that includes A2RU49 , P48200 , and P30532 / P32297 genes . The region was also modestly associated among never smokers . Gene expression studies confirmed the presence of P30532 /3 in lung , airway smooth muscle , and bronchial epithelial cells . A single-nucleotide polymorphism in Q13639 , a gene previously related to Q99581 (1)/FVC , achieved genome-wide statistical significance in combined meta-analysis . Top single-nucleotide polymorphisms in Q9H013 , P10826 , O14495 , and Q8TE59 were nominally replicated in the P48444 meta-analysis . CONCLUSIONS : These results suggest an important role for the P30532 /3 region as a genetic risk factor for airflow obstruction that may be independent of smoking and implicate the Q13639 gene in the etiology of airflow obstruction . DB00169 derivatives with adamantane or lactone ring side chains are cell type-selective vitamin D receptor modulators . The vitamin D receptor ( P11473 ) mediates the biological actions of 1,25-dihydroxyvitamin D(3) [ 1,25(OH)(2)D(3) ] , the active form of vitamin D , which regulates calcium homeostasis , immunity , cellular differentiation , and other physiological processes . We investigated the effects of three 1,25(OH)(2)D(3) derivatives on P11473 function . AD47 has an adamantane ring and LAC67a and LAC67b have lactone ring substituents at the side chain position . These vitamin D derivatives bind to P11473 but do not stabilize an active cofactor conformation . In a P11473 transfection assay , AD47 and LAC67b act as partial agonists and all three compounds inhibit P11473 activation by 1,25(OH)(2)D(3) . The derivatives enhanced the heterodimerization of P11473 with the retinoid X receptor , an effect unrelated to agonist/antagonist activity . AD47 and LAC67b weakly induced recruitment of the Q15788 cofactor to P11473 , and all three derivatives inhibited the recruitment of P52701 family cofactors to P11473 induced by 1,25(OH)(2)D(3) . It is noteworthy that AD47 induced Q15648 recruitment as effectively as 1,25(OH)(2)D(3) , whereas LAC67a and LAC67b were not effective . We examined the expression of endogenous P11473 target genes and the nuclear protein levels of P11473 and cofactors in several cell lines , including cells derived from intestine , bone , and monocytes , and found that the vitamin D(3) derivatives act as cell type-selective P11473 modulators . The data indicate that side chain modification is useful in the development of P11473 antagonists and tissue-selective modulators . Further elucidation of the molecular mechanisms of action of selective P11473 modulators will be essential for their clinical application . Association between IFNA genotype and the risk of sarcoidosis . Sarcoidosis is known to be a systemic granulomatous disorder characterized by a cell-mediated Th1-type inflammatory response . To identify a key genetic factor in the pathogenesis of sarcoidosis , we investigated single nucleotide polymorphisms within 10 candidate genes involved in type 1 immune process ( P01571 , P01574 , P01579 , P15260 , P38484 , P29460 , P42701 , Q99665 , P25101 -1 , and P49279 ) in an association-based study of 102 Japanese patients with sarcoidosis , 114 with tuberculosis , and 110 control subjects . After correction for multiple testing , an P01571 polymorphism ( 551T --> G ) was found to be associated with susceptibility to sarcoidosis ( odds ratio 3.27 [ 95 % CI : 1.44-7.46 ] , P=0.004 , P(c)=0.04 ) , but not to tuberculosis . We observed no significant associations with the other polymorphisms of the Th1-related genes . We further typed another IFNA polymorphism ( P01566 60T --> A ) and confirmed two major haplotypes of the IFNA gene , viz. , allele 1 : P01566 [ 60T ] - P01571 [ 551T ] and allele 2 : P01566 [ 60A ] - P01571 [ 551G ] , in the Japanese population . In healthy subjects , IFNA allele 2 , which is over-represented in patients with sarcoidosis , was significantly associated with increased IFN-alpha and IL-12p70 production induced by Sendai virus in vitro . This study suggests that possession of the IFNA allele with higher levels of IFN-alpha significantly increases the risk of sarcoidosis . A hemolytic assay for the estimation of functional mannose-binding lectin levels in human serum . A simple assay was developed to estimate functional mannose-binding lectin ( P11226 ) levels in serum based on the principle of yeast-induced bystander lysis of chicken erythrocytes ( ChE ) . The assay is sensitive to inhibition by ethylene glycol bis- ( beta-aminoethyl ether ) -N,N,N',N'-tetraacetic acid ( EGTA ) ( which allows alternative pathway activation ) , ethylene diamine tetraacetic acid ( DB00974 ) , mannose , N-acetylglucosamine and P05155 ( DB05341 ) , whereas it was not inhibited by galactose . A high-titer human anti-mannan antibody-containing serum with 0.06 microg P11226 /ml gave a functional signal corresponding to 0.12 microg equivalents P11226 /ml , indicating that anti-mannan antibodies are poorly hemolytic in the assay . The assay is well suited for the large-scale testing of patient samples for a functional P11226 pathway of complement activation . PEGylation of growth hormone-releasing hormone ( P01286 ) analogues . Synthetically produced GRF1-29 ( DB00010 ) has an amino acid composition identical to the N-terminal 29 amino acids sequence of the natural hypothalamic GHRH1-44 ( Figure 1 ) . It maintains bioactivity in vitro and is almost equally effective in eliciting secretion of endogenous growth hormone in vivo . The main drawbacks associated with the pharmaceutical use of hGRF1-29 relate to its short half-life in plasma , about 10-20 min in humans , which is caused mostly by renal ultrafiltration and enzymatic degradation at the N terminus . PEGylation has been considered as one valid approach to obtain more stable forms of the peptide , with a longer in vivo half-life and ultimately with increased pharmacodynamic response along the somatotropic axis ( endogenous GH , DB01277 levels ) . Different PEGylated P01286 conjugates were obtained and their bioactivity was tested in vitro and in vivo by monitoring endogenous growth hormone ( GH ) serum levels after intravenous ( i.v. ) injection in rats , and intravenous and subcutaneous ( s.c. ) injection in pigs . It was found that P01286 -PEG conjugates are able to bind and activate the human Q02643 , although with different potency . The effect of PEG molecular weight , number of PEG chains bound and position of PEGylation site on P01286 activity were investigated . Mono-PEGylated isomers with a PEG5000 polymer chain linked to Lys 12 or Lys 21 residues , showed high biological activity in vitro , which is similar to that of hGRF1-29 , and a higher pharmacodynamic response as compared to unmodified P01286 molecule . Activity of retinoic acid receptor-gamma selectively binding retinoids alone and in combination with interferon-gamma in breast cancer cell lines . Retinoids modulate several cell functions and especially inhibit the growth of a wide variety of cells including breast cancer . Retinoic acid receptor-gamma ( P13631 ) has been shown to mediate the antiproliferative activity of retinoids . To further test this hypothesis we examined the effects of different P13631 selectively binding retinoids ( CD2325 , CD2247 , CD666 and CD437 ) on breast cancer cell lines . With exception of CD2247 , all retinoids inhibited proliferation of MCF-7 , SKBR-3 , T47D and ZR-75-1 breast cancer cell lines , similar to the natural compound all-trans retinoic acid ( DB00755 ) . In addition , all 4 compounds were able to act synergistically with interferon-gamma ( P01579 ) in all breast cancer cell lines including the retinoid-resistant BT-20 and 734-B lines . In functional transactivation assays we demonstrated that only in the MCF-7 cell line , TPA-mediated AP-1 activity was suppressed only by DB00755 and CD2325 , whereas in SKBR-3 , another RA-sensitive breast cancer cell line , it was not . The synergistic antiproliferative activity involving retinoids and P01579 could not be explained by an enhanced anti-AP-1 activity . No correlation was found between expression of RARs and cellular retinoic acid binding proteins ( CRABPs ) and antiproliferative effects of the retinoids . P13631 selectively binding retinoids are potent inhibitors of breast cancer cell proliferation , alone and in combination with P01579 . For this reason and because of a possible low toxicity , as compared with retinoic acid , we speculate that these P13631 selective binding retinoids might be of clinical importance . In vitro expression of hard metal dust ( WC-Co ) -- responsive genes in human peripheral blood mononucleated cells . Hard metals consist of tungsten carbide ( WC ) and metallic cobalt ( Co ) particles and are important industrial materials produced for their extreme hardness and high wear resistance properties . While occupational exposure to metallic Co alone is apparently not associated with an increased risk of cancer , the WC-Co particle mixture was shown to be carcinogenic in exposed workers . The in vitro mutagenic/apoptogenic potential of WC-Co in human peripheral blood mononucleated cells was previously demonstrated by us . This study aimed at obtaining a broader view of the pathways responsible for WC-Co induced carcinogenicity , and in particular genotoxicity and apoptosis . We analyzed the profile of gene expression induced in vitro by WC-Co versus control ( 24 h treatment ) in human PBMC and monocytes using microarrays . The most significantly up-regulated pathways for WC-Co treated PBMC were apoptosis and stress/defense response ; the most down-regulated was immune response . For WC-Co treated monocytes the most significantly up- and down-regulated pathways were nucleosome/chromatin assembly and immune response respectively . Quantitative RT-PCR data for a selection of the most strongly modulated genes ( P09601 , P0DMV8 , P34931 , Q12983 , O60238 , P29275 , P25713 , Q13093 , P98066 ) , and some additionally chosen apoptosis related genes ( P10415 , Q07812 , FAS , P48023 , TNFalpha ) , confirmed the microarray data after WC-Co exposure and demonstrated limited differences between the Co-containing compounds . Overall , this study provides the first analysis of gene expression induced by the WC-Co mixture showing a large profile of gene modulation and giving a preliminary indication for a hypoxia mimicking environment induced by WC-Co exposure . P08246 inhibitors as treatment for P48444 . Chronic obstructive pulmonary disease , characterised by a slowly progressive , irreversible airways limitation , is a major worldwide cause of chronic morbidity and mortality . The imbalance between human neutrophil elastase and endogenous antiproteases may cause excess human neutrophil elastase in pulmonary tissues , which may be considered a major pathogenic factor in chronic obstructive pulmonary disease . Great effort has been devoted to finding a method to restore the balance , resulting in the discovery of potent two-typed small-molecular-weight human neutrophil elastase inhibitors . In the application of chronic obstructive pulmonary disease therapy , the human neutrophil elastase inhibitors mainly focused upon include ONO-5046 , MR-889 , L-694,458 , CE-1037 , GW-311616 and TEI-8362 as the acyl-enzyme inhibitors ; and DB03925 , AE-3763 , FK-706 , ICI-200,880 , ZD-0892 and ZD-8321 as the transition-state inhibitors . In this review , various problems that remain to be solved in the clinical use of human neutrophil elastase inhibitors are discussed . DB08895 . DB08895 ( CP-690,550 ; CP-690550 ; CP690550 ) , an orally active immunosuppressant , is being developed by Pfizer for the treatment of rheumatoid arthritis , inflammatory bowel disease , dry eyes , ankylosing spondylitis , psoriasis , psoriatic arthritis , and for the prevention of transplant rejection . DB08895 specifically inhibits Janus activated kinase 3 ( P52333 ) , which has a pivotal role in cytokine signal transduction that governs lymphocyte survival , proliferation , differentiation , and apoptosis . This review discusses the key development milestones and therapeutic trials of this drug . Niflumic acid and MSI-2216 reduce P01375 -induced mucin expression in human airway mucosa . BACKGROUND : Human chloride channel , calcium-activated 1 ( A8K7I4 ) has been shown to induce mucin ( MUC ) gene expression and mucus production in bronchial epithelial cells . Objective To investigate whether blocking A8K7I4 decreases mucus production . METHODS : Expression of A8K7I4 and mucus was stimulated with P01375 in human upper airway mucosal explant tissue . P98088 mRNA and mucus protein expression was blocked by inhibiting A8K7I4 by using channel blockers ( niflumic acid [ DB04552 ] and MSI-2216 ) without and with P01375 stimulation . Expression of P98088 , A8K7I4 , and P35354 mRNA was quantified by using real-time PCR . Mucus protein was assessed by periodic acid Schiff staining . Laser capture microdissection was performed to quantify A8K7I4 and P98088 mRNA expression in epithelial cells derived from mucosal explant tissue . RESULTS : P01375 significantly increased P98088 and A8K7I4 mRNA as well as mucus and A8K7I4 protein expression in the mucosal explant tissue ( P < .05 ) . Inhibition of A8K7I4 with DB04552 or MSI-2216 showed a significant dose-dependent reduction of mucus production for both blockers in the mucosal explant tissue ( P < .05 ) . P98088 mRNA was also decreased by both blockers in the whole mucosal tissue and in laser-captured mucosa epithelial cells . CONCLUSIONS : Unstimulated and P01375 -induced mucin expression could be decreased by DB04552 and MSI-2216 . Inhibiting A8K7I4 may be a potential new approach to reduce mucus overproduction . Refined chromosomal localization of the mismatch repair and hereditary nonpolyposis colorectal cancer genes P43246 and P52701 . The genomic loci for the mismatch repair genes P43246 and P52701 were mapped by fluorescence in situ hybridization , analysis of radiation hybrid panel markers , and linkage analysis of syntenic chromosome regions between human and mouse . Both genes were localized to chromosome 2p21 , adjacent to the luteinizing hormone/choriogonadotropin receptor gene ( P22888 ; 2p21 ) , telomeric to the D2S123 polymorphic marker , and centromeric to the calmodulin-2 gene ( CALM-2 ; 2p22-21 ) and son-of-sevenless gene ( SOS ; 2p22-21 ) . The genomic locations of P43246 and P52701 appears to be within 1 Mb of each other because they could not be separated by interphase fluorescence in situ hybridization . These results clarify the position of the chromosome 2 hereditary nonpolyposis colorectal cancer locus , which was originally reported to be associated with an adjacent region ( chromosome 2p14-16 ) . Novel synthesis of various orthogonally protected Cα-methyllysine analogues and biological evaluation of a vapreotide analogue containing ( S ) -α-methyllysine . Prochiral malonic diesters containing a quaternary carbon center have been successfully transformed into a diverse set of (t)Boc-Fmoc-α(2,2)-methyllysine-OH analogues through chiral malonic half-ester intermediates obtained via enzymatic ( Pig Liver Esterase , PLE ) hydrolysis . The variety of chiral half-ester intermediates , which vary from 1 to 6 methylene units in the side chain , are achieved in moderate to high optical purity and in good yields . The PLE hydrolysis of malonic diesters with various side chain lengths appears to obey the Jones 's PLE model according to the stereochemical configurations of the resulting chiral half-esters . The established synthetic strategy allows the construction of both enantiomers of α(2,2)-methyllysine analogues , and a ( S ) -β(2,2)-methyllysine analogue from a common synthon by straightforward manipulation of protecting groups . Two different straightforward and cost effective synthetic strategies are described for the synthesis of α(2,2)-methyllysine analogues . The described strategies should find significant usefulness in preparing novel peptide libraries with unnatural lysine analogues . A DB04894 analogue incorporating ( S ) -α(2,2)-methyllysine was prepared . However , the DB04894 analogue with ( S ) -α-methyl-α-lysine is found to lose its specific binding to somatostatin receptor subtype 2 ( P30874 ) . Requirement for distinct Janus kinases and P35610 proteins in T cell proliferation versus P01579 production following IL-12 stimulation . While IL-12 is known to activate O60674 and P29597 and induce the phosphorylation of Q14765 and P40763 , little is known regarding how the activation of these signaling molecules is related to the biologic effects of IL-12 . Using an IL-12-responsive T cell clone ( 2D6 ) , we investigated their requirements for proliferation and P01579 production of 2D6 cells . 2D6 cells could be maintained with either IL-12 or P60568 . 2D6 lines maintained with IL-12 ( 2D6(IL-12) ) or P60568 ( 2D6( P60568 ) ) exhibited comparable levels of proliferation , but produced large or only small amounts of P01579 , respectively , when restimulated with IL-12 after starvation of either cytokine . 2D6(IL-12) induced P29597 and Q14765 phosphorylation . In contrast , their phosphorylation was marginally induced in 2D6( P60568 ) . The reduced Q14765 phosphorylation was due to a progressive decrease in the amount of Q14765 protein along with the passages in P60568 -containing medium . 2D6(IL-12) and 2D6( P60568 ) similarly proliferating in response to IL-12 induced comparable levels of O60674 activation and P42229 phosphorylation . O60674 was associated with P42229 , and IL-12-induced P42229 phosphorylation was elicited in the absence of P52333 activation . These results indicate that IL-12 has the capacity to induce/maintain Q14765 and P42229 proteins , and that P29597 and O60674 activation correlate with Q14765 phosphorylation/ P01579 induction and P42229 phosphorylation/cellular proliferation , respectively .

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interacts_with DB00203?

multiple_choice

Transporters in cholelithiasis . Gallstones are a common and costly disease with a projected increase in prevalence due to the increasing ageing population . Numerous endogenous and environmental factors are aetiologically related to this multifactorial disease , and genetic studies continue to unravel the pathobiological mechanisms related to gallstone formation . In particular , variants of genes encoding hepatobiliary transporters have been implicated in gallstone disease and , given their ability to influence biliary lipid composition , have undergone considerable investigation . Here we summarize the role of enterohepatic transporters in cholelithogenesis with a particular focus on pertinent DB00171 -binding cassette transporters ( P21439 , O95342 , P13569 , and Q9H222 / Q9UBA6 ) . Expression cloning screen for modifiers of amyloid precursor protein shedding . Ectodomain shedding of the amyloid precursor protein ( P05067 ) is a key regulatory step in the generation of the amyloid beta peptide ( Abeta ) , which is thought to provoke the pathogenesis of Alzheimer 's disease . To better understand the cellular processes that regulate ectodomain shedding of P05067 we used human embryonic kidney 293 cells and applied a sib-selection expression cloning approach . In addition to a known activator of P05067 shedding -- protein kinase A -- the following cDNAs were identified : the endocytic proteins endophilin A1 and A3 , the metabotropic glutamate receptor 3 ( Q14832 ) , palmitoyl-protein thioesterase 1 ( P50897 ) , Numb-like and the kinase Q9Y2U5 . Endophilins A1 and A3 , as well as Q14832 activated P05067 shedding relatively specifically . They had little or no effect on the shedding of the unrelated membrane proteins P01375 receptor 2 and P16109 glycoprotein ligand-1 . In contrast , Q9Y2U5 and PKA also increased shedding of P01375 receptor 2 , suggesting that these kinases contribute to a general program regulating ectodomain shedding . The strongest activator of P05067 shedding , endophilin A3 , reduced the rate of P05067 endocytosis and specifically increased P05067 shedding by the protease alpha-secretase , as measured in an antibody uptake assay and by immunoblot analysis . This suggests that endophilin A3 is a novel modulator of P05067 trafficking affecting access of P05067 to alpha-secretase . In summary , this study shows that expression cloning is a suitable way to identify proteins controlling ectodomain shedding of membrane proteins . Further evidence for a functional role of the glutamate receptor gene Q14832 in schizophrenia . In recent years , evidence has been accumulating indicating a major role of glutamate in the pathogenesis and pathophysiology of schizophrenia . Of particular importance in this regard are the metabotropic glutamate receptors ( GRM ) . Thus , a recently published trial of the amino acid analogue DB05096 , which exerts its effects through the activation of the glutamate receptors Q14832 / Q14416 , showed an improvement of positive and negative symptoms comparable to treatment with olanzapine . A functional variant of Q14832 has been described which modulates synaptic glutamate levels . We assessed whether this functional variant rs6465084 is related to schizophrenia in a large sample of patients and controls . We found an increased frequency of the A allele ( p=0.027 ) and the AA genotype ( p=0.024 ) in schizophrenia patients . Moreover , in an assessment of schizophrenia endophenotypes , patients of the AA genotype performed poorly in the digit symbol test , a measure of attention ( p=0.008 ) . Our results provide further evidence for the potential importance of the glutamate receptor Q14832 in schizophrenia , and indicate that the novel antipsychotic DB05096 may actually be targeting a pathogenic pathway of schizophrenia . DB04540 and plant sterol absorption : recent insights . The recent discovery of transporters in the intestinal mucosa and the canalicular membrane has given new insights into the regulation of intestinal absorption as well as the biliary output of cholesterol and plant sterols . The 2 adenosine triphosphate ( DB00171 ) -binding cassette ( DB01048 ) half-transporters Q9H222 and Q9H221 are expressed in the mucosa cells and the canalicular membrane , and they resecrete sterols , especially absorbed plant sterols , back into the intestinal lumen and from the liver into bile . Defects of either of these cotransporters lead to the rare inherited disease of phytosterolemia , which is clinically defined by hyperabsorption and diminished biliary excretion of plant sterols . Furthermore , it has been recently demonstrated that the Niemann-Pick C1-Like 1 ( Q9UHC9 ) transporter is most likely responsible for the transport of cholesterol and plant sterols from the brush border membrane into the intestinal mucosa . DB00973 interferes with Q9UHC9 , reducing the intestinal uptake of cholesterol and plant sterols . These new findings contribute to our understanding of cholesterol and plant sterol concentrations in serum , and the effect of dietary and drug intervention to reduce serum concentrations of sterols . PP2Cdelta ( Ppm1d , O15297 ) , an endogenous inhibitor of p38 MAPK , is regulated along with Trp53 and Cdkn2a following p38 MAPK inhibition during mouse preimplantation development . Preimplantation embryos utilize mitogen-activated protein kinase signaling ( MAPK ) pathways to relay signals from the external environment to prepare appropriate responses and adaptations to a changing milieu . It is therefore important to investigate how MAPK pathways are regulated during preimplantation development . This study was conducted to investigate whether PP2Cdelta ( Ppm1d , O15297 ) is expressed during mouse preimplantation development and to determine the influences of p38 MAPK inhibition on expression of Trp53 ( p53 ) , Ppm1d , ( O15297 ) , and Cdkn2a ( p16 ) during mouse preimplantation development . Our results indicate that Trp53 , Ppm1d , and Cdkn2a mRNAs and TRP53 and PP2Cdelta proteins are expressed throughout mouse preimplantation development . Treatment of 2-cell embryos with DB04338 ( potent inhibitor of p38 MAPK alpha/beta/ Q16539 /11 ) significantly increased Trp53 , Ppm1d and Cdkn2a and Mapk14 mRNA levels at 12 and 24 hr . Treatment of 8-cell embryos with DB04338 for 12 hr increased Trp53 , Ppm1d , and Cdkn2a mRNA levels , but not Mapk14 mRNA levels . Treatment of 8-cell embryos for 24 hr increased Trp53 , and Ppm1d mRNA levels , but decreased Cdkn2a and Mapk14 mRNA levels . Therefore , blockade of p38 MAPK activity is associated with embryo stage specific influences on Trp53 , Ppm1d , Cdkn2a , and Mapk14 expression during mouse preimplantation development . These results define downstream targets of p38 MAPK during preimplantation development and indicate that the p38 MAPK pathway regulates Trp53 , Ppm1d , and Cdkn2a expression . This study increases our understanding of the mechanisms controlling preimplantation development and of the interactions between preimplantation embryos and their culture environments . Estrogen stimulates Th2 cytokine production and regulates the compartmentalisation of eosinophils during allergen challenge in a mouse model of asthma . BACKGROUND : The observation that asthma becomes more prevalent following puberty in females suggests estrogen potentiates the development of this disease . However , most studies examining the role of estrogen in rodent models of asthma are complicated by their reliance on ovariectomised mice in which hormones other than estrogen are also attenuated . METHODS : We aimed to understand the influence of estrogen on allergic airway disease by using type I ( tamoxifen ) or type II ( DB00947 ) antagonists in female mice or delivering estradiol to male mice during aeroallergen challenge . RESULTS : The antagonists showed that estrogen promoted both the mobilisation of bone marrow eosinophils and egression of eosinophils to the airway lumen . These findings were corroborated in male mice treated with estradiol , which increased eosinophil numbers in both blood and airways . Estrogen stimulated goblet cell hyperplasia and baseline lung resistance , but had little effect on the number of eosinophils in the bronchial submucosa or methacholine-induced airway hyperreactivity . P03372 α was expressed by P01730 + T cells from allergic mice , and estrogen promoted the production of P05113 and P35225 , and suppressed the production of the eicosanoid 12-HETE by mediastinal lymph node cells . CONCLUSIONS : These data show that during aeroallergen challenge , estrogen stimulates Th2 cytokine production , which may be linked to its ability to suppress 12-HETE . Lung resistance at baseline , goblet cell hyperplasia and the compartmentalisation of eosinophils was also influenced by estrogen . However , estrogen does not play a major role in stimulating enhanced sensitivity to methacholine-induced lung resistance . DB00203 inhibits calcineurin/ Q13469 -mediated cyclin A expression in pulmonary artery smooth muscle cells . AIMS : To examine whether calcineurin/NFAT signaling pathway leads to proliferation of pulmonary artery smooth muscle cells ( PASMCs ) by regulating cell cycle proteins and whether the phosphodiesterase-5 ( O76074 ) inhibitor sildenafil affects calcineurin/NFAT-induced cell proliferation . MAIN METHODS : A [(3)H]thymidine incorporation assay was used to examine DNA synthesis ( cell proliferation ) ; cyclin A and Q13469 expressions were determined by Western blot . P24941 ( P24941 ) activity was measured with an in vitro kinase activity assay , and calcineurin and NFAT activity were evaluated using a calcineurin assay kit and a luciferase activity assay , respectively . A chemical inhibitor or siRNA transfection was used to inhibit calcineurin/NFAT signaling pathway . KEY FINDINGS : Serotonin dose-dependently stimulated cyclin A expression in PASMCs . This effect was accompanied by dose-dependent increases in P24941 activity and the rate of DNA synthesis . At the same time , PASMCs treated with serotonin showed dose-dependent activation of calcineurin/NFAT signaling pathway . Inhibition of calcineurin activity by cyclosporine A or loss of Q13469 protein by siRNA transfection abolished serotonin-induced cyclin A expression and consequent P24941 activation and DNA synthesis . We further found that pretreatment of cells with sildenafil suppressed serotonin-triggered activation of calcineurin/ Q13469 signaling pathway and resultant cyclin A expression , P24941 activation and cell proliferation , while the presence of DT-3 [ a specific protein kinase G ( PKG ) peptide inhibitor ] reversed the effects of sildenafil on PASMCs . SIGNIFICANCE : Our study suggests that enhanced PKG activity suppresses calcineurin/ Q13469 cascade-mediated cyclin A expression , P24941 activation and PASMC proliferation to contribute to the overall effects of sildenafil in the treatment of pulmonary hypertension . FIPSDock : a new molecular docking technique driven by fully informed swarm optimization algorithm . The accurate prediction of protein-ligand binding is of great importance for rational drug design . We present herein a novel docking algorithm called as FIPSDock , which implements a variant of the Fully Informed Particle Swarm ( FIPS ) optimization method and adopts the newly developed energy function of AutoDock 4.20 suite for solving flexible protein-ligand docking problems . The search ability and docking accuracy of FIPSDock were first evaluated by multiple cognate docking experiments . In a benchmarking test for 77 protein/ligand complex structures derived from GOLD benchmark set , FIPSDock has obtained a successful predicting rate of 93.5 % and outperformed a few docking programs including particle swarm optimization ( Q9P0Z9 )@AutoDock , SODOCK , AutoDock , DOCK , Glide , GOLD , FlexX , Surflex , and MolDock . More importantly , FIPSDock was evaluated against Q9P0Z9 @AutoDock , SODOCK , and AutoDock 4.20 suite by cross-docking experiments of 74 protein-ligand complexes among eight protein targets ( P24941 , P03372 , F2 , Q16539 , P22894 , P45452 , Q07343 , and O76074 ) derived from Sutherland-crossdock-set . Remarkably , FIPSDock is superior to Q9P0Z9 @AutoDock , SODOCK , and AutoDock in seven out of eight cross-docking experiments . The results reveal that FIPS algorithm might be more suitable than the conventional genetic algorithm-based algorithms in dealing with highly flexible docking problems . A cell-intrinsic inhibitor of HIV-1 reverse transcription in P01730 (+) T cells from elite controllers . HIV-1 reverse transcription represents the predominant target for pharmacological inhibition of viral replication , but cell-intrinsic mechanisms that can block HIV-1 reverse transcription in a clinically significant way are poorly defined . We find that effective HIV-1 reverse transcription depends on the phosphorylation of viral reverse transcriptase by host cyclin-dependent kinase ( CDK ) 2 at a highly conserved DB00156 residue . P24941 -dependent phosphorylation increased the efficacy and stability of viral reverse transcriptase and enhanced viral fitness . Interestingly , P38936 , a cell-intrinsic CDK inhibitor that is upregulated in P01730 (+) T cells from " elite controllers , " potently inhibited P24941 -dependent phosphorylation of HIV-1 reverse transcriptase and significantly reduced the efficacy of viral reverse transcription . These data suggest that P38936 can indirectly block HIV-1 reverse transcription by inhibiting host cofactors supporting HIV-1 replication and identify sites of viral vulnerability that are effectively targeted in persons with natural control of HIV-1 replication . Oral leucine supplementation is sensed by the brain but neither reduces food intake nor induces an anorectic pattern of gene expression in the hypothalamus . DB00149 activates the intracellular mammalian target of the rapamycin ( P42345 ) pathway , and hypothalamic P42345 signaling regulates food intake . Although central infusion of leucine reduces food intake , it is still uncertain whether oral leucine supplementation is able to affect the hypothalamic circuits that control energy balance . We observed increased phosphorylation of p70s6k in the mouse hypothalamus after an acute oral gavage of leucine . We then assessed whether acute oral gavage of leucine induces the activation of neurons in several hypothalamic nuclei and in the brainstem . DB00149 did not induce the expression of Fos in hypothalamic nuclei , but it increased the number of Fos-immunoreactive neurons in the area postrema . In addition , oral gavage of leucine acutely increased the 24 h food intake of mice . Nonetheless , chronic leucine supplementation in the drinking water did not change the food intake and the weight gain of ob/ob mice and of wild-type mice consuming a low- or a high-fat diet . We assessed the hypothalamic gene expression and observed that leucine supplementation increased the expression of enzymes ( P54687 , O15382 and O14874 ) that metabolize branched-chain amino acids . Despite these effects , leucine supplementation did not induce an anorectic pattern of gene expression in the hypothalamus . In conclusion , our data show that the brain is able to sense oral leucine intake . However , the food intake is not modified by chronic oral leucine supplementation . These results question the possible efficacy of leucine supplementation as an appetite suppressant to treat obesity . DB00203 promotes adipogenesis through a PKG pathway . DB00203 is the first oral O76074 inhibitor for the treatment of erectile dysfunction and pulmonary arterial hypertension . In the present study , we investigated the effect of sildenafil on adipogenesis in 3T3L1 preadipocytes . Treatment with sildenafil for 8 days significantly promoted adipogenesis characterized by increased lipid droplet and triglyceride content in 3T3L1 cells . Meanwhile , sildenafil induced a pronounced up-regulation of the expression of adipocyte-specific genes , such as aP2 and P14672 . The results by RT-PCR and Western blotting further showed that sildenafil increased the sequential expression of P17676 , Q07869 gamma and P49715 . Additionally , we found that the other two O76074 inhibitors ( vardenafil and tadalafil ) and the cGMP analog 8-pCPT-cGMP also increased adipogenesis . Likewise , 8-pCPT-cGMP could up-regulate the expression of adipogenic and adipocyte-specific genes . Importantly , the PKG inhibitor Rp-8-pCPT-cGMP was able to inhibit both sildenafil and 8-pCPT-cGMP-induced adipogenesis . Furthermore , sildenafil promoted basal and insulin-mediated glucose uptake in 3T3L1 cells , which was counteracted by Rp-8-pCPT-cGMP . These results indicate that sildenafil could promote adipogenesis accompanied by increased glucose uptake through a PKG pathway at least partly . Anti- P05231 -receptor-alpha ( tocilizumab ) does not inhibit human monocyte-derived dendritic cell maturation or alloreactive T-cell responses . Significant comorbidites and lethality complicate GVHD and its treatment . Targeting the cytokine milieu may improve GVHD control ; and P05231 is an attractive candidate , given its role in dendritic cell activation and T-cell differentiation . DB06273 is a humanized mAb to P05231 -receptor-α ( P08887 -α ) , which is Food and Drug Administration-approved for treatment of rheumatoid arthritis . Mouse transplant models have demonstrated that P05231 blockade also improves GVHD scores and survival . Definitive immunologic effects of P05231 inhibition have not emerged given inconsistent alterations in regulatory T cells ( Tregs ) and suppression of T-cell proliferation . Despite on-target suppression of P08887 -α signaling in human monocyte-derived dendritic cells ( moDCs ) and T cells , our data show no effect on moDC maturation/activation , alloreactive T-cell proliferation , Treg expansion , or allogeneic Th1/Th17 responses in vitro . These findings merit attention in any clinical trials of tocilizumab for GVHD prevention or treatment and provide a rationale for evaluating more specific inhibitors of downstream O60674 / P40763 signaling as well . [ Effect of shu di-huang on the transmitter and receptor of amino acid in brain and learning and memory of dementia model ] . OBJECTIVE : To observe the mechanism of SHU-Dihuang on the function of learning and memory . METHOD : On the dementia model mouse caused by AlCl3 and the rats model damaged thalamic arcuate nucleus with MSG , we observed the function of learning and memory by step down task and Morris water maze task , mensurated the content of glutamic acid and gamma-aminobutyric acid by TLC , and observed the expression of Q05586 and GABAR in hippocampi by immunohistochemical means . RESULT : Shu Di Huang could decrease the times of mistakes and prolong the incubation period in step down task , and shorten the incubation period of seeking the platform in Morris water maze task . Shu Di Huang could adjust the content of DB00142 and GABA in brain , and increase the expression of hippocampal Q05586 and GABAR as well . CONCLUSION : Shu Di Huang can improve the function of learning and memory of dementia animal model , and its mechanism may be related to the adjustment of the content of DB00142 and GABA in brain , and increase of the expression of hippocampal Q05586 and GABAR . O76074 inhibitors enhance celecoxib killing in multiple tumor types . The present studies determined whether clinically relevant phosphodiesterase 5 ( O76074 ) inhibitors interacted with a clinically relevant NSAID , celecoxib , to kill tumor cells . Celecoxib and O76074 inhibitors interacted in a greater than additive fashion to kill multiple tumor cell types . Celecoxib and sildenafil killed ex vivo primary human glioma cells as well as their associated activated microglia . Knock down of O76074 recapitulated the effects of O76074 inhibitor treatment ; the nitric oxide synthase inhibitor L-NAME suppressed drug combination toxicity . The effects of celecoxib were P35354 independent . Over-expression of O15519 -s or knock down of CD95/ Q13158 significantly reduced killing by the drug combination . CD95 activation was dependent on nitric oxide and ceramide signaling . CD95 signaling activated the JNK pathway and inhibition of JNK suppressed cell killing . The drug combination inactivated P42345 and increased the levels of autophagy and knock down of Beclin1 or Q9H1Y0 strongly suppressed killing by the drug combination . The drug combination caused an ER stress response ; knock down of IRE1α/ P17861 enhanced killing whereas knock down of eIF2α/ P18848 / P35638 suppressed killing . DB00203 and celecoxib treatment suppressed the growth of mammary tumors in vivo . Collectively our data demonstrate that clinically achievable concentrations of celecoxib and sildenafil have the potential to be a new therapeutic approach for cancer . PPARgamma activation abolishes LDL-induced proliferation of human aortic smooth muscle cells via SOD-mediated down-regulation of superoxide . Native LDL would be a mitogenic and chemotactic stimulus of VSMC proliferation and differentiation in the atherosclerotic lesion where endothelial disruption occurred . In previous studies , our group investigated the molecular mechanisms by which LDL induces P10145 production and by which PPARalpha activation abolishes LDL effects in human aortic SMCs ( hAoSMCs ) . Herein is the first report of PPARgamma activation by troglitazone ( TG ) exerting its inhibitory effects on LDL-induced cell proliferation via generation not of H(2)O(2) , but of O2(.-) , and the subsequent activation of Erk1/2 in hAoSMCs . Moreover , in this study TG abolished the LDL-accelerated G(1)-S progression to control levels via down-regulation of active cyclinD1/ P11802 and cyclinE/ P24941 complexes and up-regulation of P38936 (Cip1) expression . TG exerted its anti-proliferative effects through the up-regulation of basal superoxide dismutase ( SOD ) expression . This data suggests that the regulation of O2(.-) is located at the crossroads between LDL signaling and cell proliferation . p35 deficiency accelerates HMGB-1-mediated neuronal death in the early stages of an Alzheimer 's disease mouse model . The activities of Q00535 and p35 are thought to be important in the pathogenesis of neurodegenerative diseases , including Alzheimer 's disease ( AD ) . We studied the effect of p35 deletion in Tg2576 mice , which is an AD animal model . To obtain the desired mice , we crossed p35(-/-) with Tg2576 mice . The resulting p35(-/-)/Tg2576 ( KO/Tg ) mice displayed higher mortality rates and exhibited impaired spatial learning and memory at 6 months of age . Using immunohistochemical and biochemical approaches , we observed a reduction in the expression of pre- and post-synaptic markers such as Q05586 , synaptophysin and GluR1 . In addition , the intensity of P11137 -positive dendrites extending from neuronal cell bodies was significantly decreased in KO/Tg mice compared with KO/WT and WT/Tg mice . We also detected increased neuronal cell death in the hippocampus , along with thinned and collapsed morphological changes in the alveus region and a dramatic increase in the number of microglial cells . Microglial infiltration in the hippocampus could result in the increased secretion of the soluble high mobility group box-1 protein ( HMGB-1 ) . The secretion of HMGB-1 is increased by Aβ , and secretion of HMGB-1 promotes neuronal cell death . Moreover , we found that HMGB-1 secretion induced by Aβ in KO/Tg mice gave rise to ER-mediated cell death . In summary , during the stages of KO/Tg mice model , the microglial infiltration and secretion of soluble HMGB-1 were significantly increased in the hippocampus . These conditions promote neuronal death , synaptic destruction and behavioral deficits . SAR and biological evaluation of analogues of a small molecule histone deacetylase inhibitor N-(2-aminophenyl)-4-((4-(pyridin-3-yl)pyrimidin-2-ylamino)methyl)benzamide ( DB05651 ) . Analogues of the clinical compound DB05651 ( A ) were designed and synthesized . These compounds inhibit recombinant human Q13547 with IC(50) values in the sub-micromolar range . In human cancer cells growing in culture these compounds induce hyperacetylation of histones , cause expression of the tumor suppressor protein P38936 ( P38936 /CIP1) , and inhibit cellular proliferation . Lead molecule of the series , compound 25 is metabolically stable , possesses favorable pharmacokinetic characteristics and is orally active in vivo in different mouse tumor xenograft models . DB00203 attenuates inflammation and oxidative stress in pelvic ganglia neurons after bilateral cavernosal nerve damage . Erectile dysfunction is a common complication for patients undergoing surgeries for prostate , bladder , and colorectal cancers , due to damage of the nerves associated with the major pelvic ganglia ( P29372 ) . Functional re-innervation of target organs depends on the capacity of the neurons to survive and switch towards a regenerative phenotype . O76074 inhibitors ( PDE5i ) have been successfully used in promoting the recovery of erectile function after cavernosal nerve damage ( BCNR ) by up-regulating the expression of neurotrophic factors in P29372 . However , little is known about the effects of PDE5i on markers of neuronal damage and oxidative stress after BCNR . This study aimed to investigate the changes in gene and protein expression profiles of inflammatory , anti-inflammatory cytokines and oxidative stress related-pathways in P29372 neurons after BCNR and subsequent treatment with sildenafil . Our results showed that BCNR in Fisher-344 rats promoted up-regulation of cytokines ( interleukin- 1 ( IL-1 ) β , P05231 , P22301 , transforming growth factor β 1 ( TGFβ1 ) , and oxidative stress factors ( DB02701 adenine dinucleotide phosphate ( NADPH ) oxidase , P05164 ( P05164 ) , inducible nitric oxide synthase ( P35228 ) , P01375 receptor superfamily member 5 ( P25942 ) that were normalized by sildenafil treatment given in the drinking water . In summary , PDE5i can attenuate the production of damaging factors and can up-regulate the expression of beneficial factors in the P29372 that may ameliorate neuropathic pain , promote neuroprotection , and favor nerve regeneration . Phosphodiesterase-5 inhibitor sildenafil prevents neuroinflammation , lowers beta-amyloid levels and improves cognitive performance in P05067 / P49768 transgenic mice . Memory deficit is a marker of Alzheimer 's disease ( AD ) that has been highly associated with the dysfunction of cyclic GMP ( cGMP ) signaling and an ongoing inflammatory process . Phosphodiesterase-5 ( O76074 ) inhibitors prevent the breakdown of cGMP and are currently studied as a possible target for cognitive enhancement . However , it is still unknown whether inhibition of O76074 reversed β-amyloid peptide ( Aβ ) -induced neuroinflammation in P05067 / P49768 transgenic ( Tg P05067 / P49768 ) mice . The present study evaluated the cognitive behaviors , inflammatory mediators , and cGMP/PKG/pCREB signaling in 15-month-old Tg P05067 / P49768 mice and age-matched wild-type ( WT ) mice that were treated with O76074 inhibitor sildenafil and the inhibitor of cGMP-dependent protein kinase Rp-8-Br-PET-cGMPS . In comparison with WT mice , Tg P05067 / P49768 mice were characterized by impaired cognitive ability , neuroinflammatory response , and down-regulated cGMP signaling . DB00203 reversed these memory deficits and cGMP/PKG/pCREB signaling dysfunction ; it also reduced both the soluble Aβ1-40 and Aβ1-42 levels in the hippocampus . These effects of sildenafil were prevented by intra-hippocampal infusion of the Rp-8-Br-PET-cGMPS . These results suggest that sildenafil could restore cognitive deficits in Tg P05067 / P49768 mice by the regulation of PKG/pCREB signaling , anti-inflammatory response and reduction of Aβ levels . Activation of intestinal peroxisome proliferator-activated receptor-α increases high-density lipoprotein production . AIMS : Peroxisome proliferator-activated receptor ( Q07869 ) -α is a transcription factor controlling lipid metabolism in liver , heart , muscle , and macrophages . Peroxisome proliferator-activated receptor-α activation increases plasma HDL cholesterol and exerts hypotriglyceridaemic actions via the liver . However , the intestine expresses Q07869 -α , produces HDL and chylomicrons , and is exposed to diet-derived Q07869 -α ligands . Therefore , we examined the effects of Q07869 -α activation on intestinal lipid and lipoprotein metabolism . METHODS AND RESULTS : The impact of Q07869 -α activation was evaluated in term of HDL-related gene expression in mice , ex vivo in human jejunal biopsies and in Caco-2/TC7 cells . Apolipoprotein-AI/HDL secretion , cholesterol esterification , and trafficking were also studied in vitro . In parallel to improving plasma lipid profiles and increasing liver and intestinal expression of fatty acid oxidation genes , treatment with the dual Q07869 -α/δ ligand DB05187 resulted in a more pronounced increase in plasma HDL compared with fenofibrate in mice . DB05187 , but not fenofibrate , increased the expression of HDL production genes such as apolipoprotein-AI and DB00171 -binding cassette A1 transporter in murine intestines . A similar increase was observed upon Q07869 -α activation of human biopsies and Caco-2/TC7 cells . Additionally , HDL secretion by Caco-2/TC7 cells increased . Moreover , Q07869 -α activation decreased the cholesterol esterification capacity of Caco-2/TC7 cells , modified cholesterol trafficking , and reduced apolipoprotein-B secretion . CONCLUSION : Peroxisome proliferator-activated receptor-α activation reduces cholesterol esterification , suppresses chylomicron , and increases HDL secretion by enterocytes . These results identify the intestine as a target organ of Q07869 -α ligands with entero-hepatic tropism to reduce atherogenic dyslipidaemia . DB00203 restores cognitive function without affecting β-amyloid burden in a mouse model of Alzheimer 's disease . BACKGROUND AND PURPOSE : Inhibitors of phosphodiesterase 5 ( O76074 ) affect signalling pathways by elevating cGMP , which is a second messenger involved in processes of neuroplasticity . In the present study , the effects of the O76074 inhibitor , sildenafil , on the pathological features of Alzheimer 's disease and on memory-related behaviour were investigated . EXPERIMENTAL APPROACH : DB00203 was administered to the Tg2576 transgenic mouse model of Alzheimer 's disease and to age-matched negative littermates ( controls ) . Memory function was analysed using the Morris water maze test and fear conditioning tasks . Biochemical analyses were performed in brain lysates from animals treated with saline or with sildenafil . KEY RESULTS : Treatment of aged Tg2576 animals with sildenafil completely reversed their cognitive impairment . Such changes were accompanied in the hippocampus by a reduction of tau hyperphosphorylation and a decrease in the activity of glycogen synthase kinase 3β ( GSK3β ) and of cyclin-dependent kinase 5 ( Q00535 ) ( p25/p35 ratio ) . Moreover , sildenafil also increased levels of brain-derived neurotrophic factor ( P23560 ) and the activity-regulated cytoskeletal-associated protein ( Arc ) in the hippocampus without any detectable modification of brain amyloid burden . CONCLUSIONS AND IMPLICATIONS : DB00203 improved cognitive functions in Tg2576 mice and the effect was not related to changes in the amyloid burden . These data further strengthen the potential of sildenafil as a therapeutic agent for Alzheimer 's disease . O76074 inhibitors for cystic fibrosis : can they also enhance chloride transport ? Evaluation of : Lubamba B , Lecourt H , Lebacq J , et al . Preclinical evidence that sildenafil and vardenafil activate chloride transport in cystic fibrosis . Am J Respir Crit Care Med 2008;177(5):506-15 . BACKGROUND : Many compounds are currently under investigation for cystic fibrosis ( CF ) , a genetic disorder caused by various mutations in cystic fibrosis transmembrane regulator ( P13569 ) . OBJECTIVE : To evaluate a preclinical study on the effects of sildenafil and vardenafil , two O76074 inhibitors , on ion transport in CF animal models . METHODS/RESULTS : The effects of sildenafil and vardenafil on chloride and sodium conductance were assessed in both DeltaF508CFTR and P13569 knockout mice models . They improved chloride transport in the DeltaF508CFTR model and had no effects in the P13569 knockout model . CONCLUSION : DB00203 and vardenafil are promising agents for CF therapy provided they also demonstrate clinical efficacy .

MH_train_1311

MH_train_1311

MH_train_1311

interacts_with DB09048?

multiple_choice

Differential effects of all-trans and 13-cis-retinoic acid on mRNA levels of nuclear retinoic acid receptors in rat lung and liver . The effects of three retinoids , all-trans-retinoic acid ( all-trans-RA ) , 13- DB00982 , and etretin were examined on mRNA abundance of nuclear retinoic acid receptors ( P10276 , beta , and gamma ) in lung and liver of retinol deficient and chow fed rats . All-trans-RA increased lung P10826 mRNA levels 5 or 11-fold in chow fed and retinol deficient rats , respectively . Similarly to lung , liver P10826 mRNA levels were 3-fold higher in retinol deficient rats fed all-trans-RA than the rats fed cottonseed oil . Lung P13631 mRNA levels were also induced 2-fold by all-trans-RA . In contrast to this , 13- DB00982 and etretin at equimolar doses failed to enhance lung or liver P10826 or lung P13631 mRNA levels in retinol deficient rats . These data for the first time show that all-trans-RA is more effective than its 13-cis-isomer in regulating the expression of P10826 and gamma transcripts in adult animal . Anti-nociceptive effect of a conjugate of DB05875 and light chain of botulinum neurotoxin type A . Neuropathic pain is a debilitating condition resulting from damage to sensory transmission pathways in the peripheral and central nervous system . A potential new way of treating chronic neuropathic pain is to target specific pain-processing neurons based on their expression of particular receptor molecules . We hypothesized that a toxin-neuropeptide conjugate would alter pain by first being taken up by specific receptors for the neuropeptide expressed on the neuronal cells . Then , once inside the cell the toxin would inhibit the neurons ' activity without killing the neurons , thereby providing pain relief without lesioning the nervous system . In an effort to inactivate the nociceptive neurons in the trigeminal nucleus caudalis in mice , we targeted the NK1 receptor ( P25103 ) using DB05875 ( SP ) . The catalytically active light chain of botulinum neurotoxin type A ( LC/A ) was conjugated with SP . Our results indicate that the conjugate DB00083 -LC:SP is internalized in cultured P25103 -expressing neurons and also cleaves the target of botulinum toxin , a component-docking motif necessary for release of neurotransmitters called P60880 . The conjugate was next tested in a murine model of DB01229 -induced neuropathic pain . An intracisternal injection of DB00083 -LC:SP decreased thermal hyperalgesia as measured by the operant orofacial nociception assay . These findings indicate that conjugates of the light chain of botulinum toxin are extremely promising agents for use in suppressing neuronal activity for extended time periods , and that DB00083 -LC:SP may be a useful agent for treating chronic pain . Expression of the tachykinin receptor mRNAs in healthy human colon . Tachykinins are a family of neuropeptides , involved in a variety of physiological and pathological processes occurring in the gastrointestinal tract . They act via three distinct types of receptors , tachykinin NK(1) , NK(2) , and NK(3) receptors , which belong to the family of G protein-coupled receptors . The aim of the present study was to characterize , for the first time in the healthy human colon , the TACR(1) , TACR(2) and TACR(3) mRNAs encoding the three different tachykinin receptors and to measure their relative expression by quantitative reverse transcription-PCR assay . Our results confirm the broad distribution of the tachykinin receptors but evidenced significant differences in the expression level of their respective mRNAs . A higher expression level of the P21452 mRNA alpha isoform , the gene encoding the functional tachykinin NK(2) receptor , was observed in comparison to P25103 and P29371 mRNAs genes encoding for NK(1) and NK(3) receptors respectively . The prevalence of the P21452 mRNA alpha isoform strongly suggests a major involvement of tachykinin NK(2) receptor in the regulation of human colonic functions . Chronic atrial fibrillation alters the functional properties of If in the human atrium . INTRODUCTION : Despite the evidence that the hyperpolarization-activated current ( If ) is highly modulated in human cardiomyopathies , no definite data exist in chronic atrial fibrillation ( cAF ) . We investigated the expression , function , and modulation of If in human cAF . METHODS AND RESULTS : Right atrial samples were obtained from sinus rhythm ( SR , n = 49 ) or cAF ( duration > 1 year , n = 31 ) patients undergoing corrective cardiac surgery . Among f-channel isoforms expressed in the human atrium ( O60741 , 2 and 4 ) , Q9Y3Q4 mRNA levels measured by RT-PCR were significantly reduced . However , protein expression was preserved in cAF compared to SR ( +85 % for Q9Y3Q4 ) ; concurrently , miR-1 expression was significantly reduced . In patch-clamped atrial myocytes , current-specific conductance ( gf ) was significantly increased in cAF at voltages around the threshold for If activation ( -60 to -80 mV ) ; accordingly , a 10-mV rightward shift of the activation curve occurred ( P < 0.01 ) . β-Adrenergic and Q13639 receptor stimulation exerted similar effects on If in cAF and SR cells , while the P01160 -mediated effect was significantly reduced ( P < 0.02 ) , suggesting downregulation of natriuretic peptide signaling . CONCLUSIONS : In human cAF modifications in transcriptional and posttranscriptional mechanisms of HCN channels occur , associated with a slight yet significant gain-of-function of If , which may contribute to enhanced atrial ectopy . Application of HapMap data to the evaluation of 8 candidate genes for pediatric slow transit constipation . BACKGROUND : Slow transit constipation ( P52823 ) affects up to 3 % of all children and is an increasingly recognized cause of chronic constipation in children . We conducted a pilot study to investigate whether genes encoding neurotransmitters ( P20366 , Q9UHF0 , P01282 , NOS1 ) and receptors ( P25103 , P21452 , P29371 , P10721 ) could be responsible for P52823 . METHODS : One hundred seventeen tag single nucleotide polymorphisms ( SNPs ) , distributed among the candidate genes , were selected from HapMap data and genotyped using Sequenom ( San Diego , CA ) technology in 35 affected families . Evaluation of association was performed by transmission disequilibrium test and multilocus analysis . RESULTS : Five SNPs ( rs3771863 , rs4580655 , rs11722288 , rs4563545 , and rs3782221 ) in the P25103 , P29371 , P10721 , and NOS1 genes were found to be potentially associated with P52823 , although the significance of these results does not withstand correction for multiple testing . CONCLUSIONS : Our data indicate that 5 SNPs in the NOS1 , P25103 , P29371 , and P10721 genes could be involved in P52823 , especially rs3771863 in intron 1 of P25103 , which showed the highest association . [ Acute myeloid leukemia. Genetic diagnostics and molecular therapy ] . Acute myeloid leukemia ( AML ) is a genetically heterogeneous disease . The genetic diagnostics have become an essential component in the initial work-up for disease classification , prognostication and prediction . More and more promising molecular targeted therapeutics are becoming available . A prerequisite for individualized treatment strategies is a fast pretherapeutic molecular screening including the fusion genes P29590 - P10276 , Q01196 - Q06455 and Q13951 - P35749 as well as mutations in the genes P06748 , P36888 and P49715 . Promising new therapeutic approaches include the combination of all- trans retinoic acid and arsentrioxid in acute promyelocytic leukemia , the combination of intensive chemotherapy with P10721 inhibitors in core-binding factor AML and P36888 inhibitors in AML with P36888 mutation , as well as gemtuzumab ozogamicin therapy in patients with low and intermediate cytogenetic risk profiles . With the advent of the next generation sequencing technologies it is expected that new therapeutic targets will be identified . These insights will lead to a further individualization of AML therapy . Characterization of Förster resonance energy transfer in a botulinum neurotoxin protease assay . Our previous article described a fluorescence-based assay for monitoring the proteolytic activity of botulinum neurotoxin types A and E ( DB00083 and BoNT/E ) . As detailed in that article , the assay is based on depolarization due to Förster resonance energy transfer between blue fluorescent protein ( Q9ULX5 ) and green fluorescent protein ( GFP ) moieties linked via residues 134-206 of P60880 ( synaptosome-associated protein of 25kDa ) , the protein substrate for DB00083 and BoNT/E . Before cleavage of this recombinant substrate , the polarization observed for the GFP emission , excited near the absorption maximum of the Q9ULX5 , is very low due to depolarization following energy transfer from Q9ULX5 to GFP . After substrate cleavage and diffusion of the fluorescent proteins beyond the energy transfer distance , the polarization is high due to observation of the emission only from directly excited GFP . This change in fluorescence polarization allows an assay , termed DARET ( depolarization after resonance energy transfer ) , that is robust and sensitive . In this article , we characterize the spectroscopic parameters of the system before and after substrate cleavage , including excitation and emission spectra , polarizations , and lifetimes . DB00171 -binding cassette transporter A1 R219K polymorphism and ischemic stroke risk in the Chinese population : a meta-analysis . Recently , many studies have been focused on the association between the DB00171 -binding cassette transporter A1 ( O95477 ) gene R219K polymorphism and ischemic stroke ( IS ) . However , the study results have been inconsistent , especially in the Chinese population . Therefore , we performed a meta-analysis to better clarify the association between the O95477 gene and IS . All of the relevant studies used in our meta-analysis were identified using PubMed , OVID , Cochrane Library , Chinese Wan Fang database , Chinese P01282 database , China National Knowledge Infrastructure ( CNKI ) , and China Biological Medicine Database ( CBM ) up to May 2013 . Statistical analysis was conducted with STATA software version 11.0 . Odds ratios with 95 % confidence intervals were applied to evaluate the strength of the association between O95477 gene R219K polymorphism and IS . Heterogeneity was evaluated using the Q-test and I(2) statistic . The funnel plots , Begg 's and Egger 's regression tests were used to assess the publication bias . Our meta-analysis showed the dominant genetic model ( OR=0.92 , 95 % CI : 0.88-0.96 ) , the recessive genetic model ( OR=0.73 , 95 % CI : 0.51-1.05 ) , the homozygote genetic model ( OR=0.64 , 95 % CI : 0.44-0.94 ) , the heterozygote genetic model ( OR=0.81 , 95 % CI : 0.69-0.95 ) , and the allelic genetic model ( OR=0.83 , 95 % CI : 0.69-0.99 ) . For R219K in IS , there were significant associations with these genetic models , but not with the recessive genetic model . Our meta-analysis indicated that the O95477 gene R219K polymorphism might be associated with IS and the K allele might be a protective factor in the Chinese population . Efficacy and safety of repeated dosing of netupitant , a neurokinin-1 receptor antagonist , in treating overactive bladder . AIM : NK-1 receptors in sensory nerves , the spinal cord and bladder smooth muscle participate in complex sensory mechanisms that regulate bladder activity . This study was designed to assess the efficacy and safety of a new P25103 antagonist , netupitant , in patients with OAB . METHODS : This was a phase II , multicenter , double-blind study in which adults with OAB symptoms > 6 months were randomized to receive 1 of 3 doses of netupitant ( 50 , 100 , 200 mg ) or placebo once daily for 8 weeks . The primary efficacy endpoint was percentage change from baseline in average number of daily micturitions at week 8 . Urinary incontinence , urge urinary incontinence ( UUI ) , and urgency episodes were also assessed . RESULTS : The primary efficacy endpoint was similar in the treatment groups ( -13.85 for placebo to -16.17 in the netupitant 200 mg group ) with no statistically significant differences between netupitant and placebo . The same was true for most secondary endpoints although a significant difference for improvement in UUI episodes and a trend for the greatest decrease in urgency episodes were seen in the netupitant 100 mg group . DB09048 was well tolerated with most treatment emergent adverse events ( AEs ) being mild . While the overall incidence of AEs increased with netupitant dose , there was no evidence for this dose dependency based on relationship to treatment , intensity , or time to onset . CONCLUSIONS : The study failed to demonstrate superiority of netupitant versus placebo in decreasing OAB symptoms , despite a trend favoring netupitant 100 mg . There were no safety concerns with daily administration of netupitant over 8 weeks . The in vitro pharmacological profile of DB05655 , a selective 5-HT(4) receptor agonist with high intrinsic activity . The in vitro pharmacological profile of DB05655 , a novel , selective 5-HT(4) receptor agonist , was compared to that of clinically efficacious gastroprokinetic 5-HT(4) receptor agonists . DB05655 produced an elevation of cyclic adenosine monophosphate in human embryonic kidney 293 cells expressing the human recombinant 5-HT(4(c)) ( h5-HT(4(c)) ) receptor ( pEC(50) = 8.3 ) and 5-HT(4) receptor-mediated relaxation of the rat esophagus ( pEC(50) = 7.9 ) and contraction of the guinea pig colon ( pEC(50) = 7.9 ) . In all in vitro assays , DB05655 was a high intrinsic activity agonist , unlike tegaserod , mosapride , and cisapride which , in the majority of test systems , had lower intrinsic activity . DB05655 had high affinity ( pK ( i ) = 7.7 ) and selectivity ( > or =25-fold ) for h5-HT(4(c)) receptors over other biogenic amine receptors . DB05655 was > 500-fold selective over other 5-HT receptors ( including h5-HT(2B) and h5-HT(3A) ) and , at 3 microM , had no effect on human ether-à-go-go-related gene K+ channels . In conclusion , DB05655 is a selective 5-HT(4) receptor agonist in vitro . The high intrinsic activity and preferential binding of DB05655 to Q13639 over other 5-HT receptors may result in an improved clinical profile for the treatment of gastrointestinal disorders of reduced motility . [ Role of neurokinin-1 receptor in lung injury in rats with acute necrotizing pancreatitis ] . OBJECTIVE : To investigate the expression of neurokinin-1 receptor ( P25103 ) in the lung tissue , and the relationship between expression of P25103 and lung injury in rats with acute necrotizing pancreatitis ( P01160 ) . METHODS : One hundred and twenty adult Sprague-Dawley rats were randomly divided into P01160 and control groups . Animals in group P01160 were induced by the retrograde intraductal infusion of 5 % sodium taurocholate ( 0.1 ml/kg ) , and animals in normal control group received laparotomy only . The accumulation of polymorphonuclear leukocytes in lung tissues was measured with myeloperoxidase ( P05164 ) assay . Lung endothelial barrier destruction was measured by lung capillary permeability ( LCP ) . Reverse transcription polymerase chain reaction ( RT-PCR ) was used to determine the mRNA expression of P25103 , western blot analysis was used to determine P25103 protein expression levels , and immunohistochemistry was used to localize expression site of P25103 . RESULTS : P25103 mRNA level was enhanced in the lung of P01160 compared with normal control group . Western blot analysis showed overexpression of P25103 protein level exited in P01160 group . Statistical analysis revealed correlation between P25103 mRNA and P05164 ( r=0.83 , P < 0.01 ) and LCP ( r=0.79 , P < 0.01 ) respectively . With immunohistochemistry staining , moderate to strong P25103 immunoreactivity was localized to alveolar membrane , I epithelium , II epithelium and polymorphonuclear leukocytes in the lung of P01160 . CONCLUSION : In P01160 , overexpression of P25103 contributes to disturbance of neuropeptides loop , resulting in aggregation of neutrophilic granulocyte and promoting deterioration of lung injury . Development of DB00644 antagonists for prostate cancer : new approaches to treatment . Prostate cancer has become the most common cancer among American men and is second only to lung cancer as a cause of male cancer-related death . Several treatment options exist for different stages of prostate cancer including observation , prostatectomy , radiation therapy , chemotherapy , and hormone therapy . Hormone therapy has evolved from the use of estrogens to gonadotropin-releasing hormone ( DB00644 ) agonists and recently , investigational DB00644 antagonists . P30968 agonists such as leuprolide , bruserelin and goserelin have been used for the treatment of prostate cancer . These agonists eventually cause the inhibition of lutenizing hormone production , which in turn causes a suppression of testosterone and dihydrotestosterone , on which continued growth of prostate cancer cells depend . Several comparative studies of leuprorelin administered as daily injections or monthly depot injections have been reported . Disease progression was prevented in more than 72 % of men administered daily leuprorelin , and in 82 % to 89 % of those receiving monthly depots . Another synthetic DB00644 analog , goserelin , has been studied in a similar population of men with daily injections producing partial responses in 60 % to 80 % of men with previously untreated prostate cancer . DB00106 , a peptide antagonist of P30968 , is also being studied for the treatment of prostate cancer . The discovery and development of DB00644 antagonists may provide an important advance for patients with prostate cancer . Clearly the studies described herein , as well as many others , outline an exciting era of research to define the optimal use of hormonal therapy in prostate cancer . Synthesis and biological activity of olomoucine II . Based on our previous experiences with synthesis of purines , novel 2,6,9-trisubstituted purine derivatives were prepared and assayed for the ability to inhibit P06493 /cyclin B kinase . One of newly synthesized compounds designated as olomoucine II , 6-[(2-hydroxybenzyl)amino]-2-[[1-(hydroxymethyl)propyl]amino]-9-isopropylpurine , displays 10 times higher inhibitory activity than roscovitine , potent and specific P06493 inhibitor . DB02116 II in vitro cytotoxic activity exceeds purvalanol A , the most potent CDK inhibitor , as it kills the CEM cells with IC(50) value of 3.0 microM . R306465 is a novel potent inhibitor of class I histone deacetylases with broad-spectrum antitumoral activity against solid and haematological malignancies . R306465 is a novel hydroxamate-based histone deacetylase ( HDAC ) inhibitor with broad-spectrum antitumour activity against solid and haematological malignancies in preclinical models . R306465 was found to be a potent inhibitor of Q13547 and -8 ( class I ) in vitro . It rapidly induced histone 3 ( H3 ) acetylation and strongly upregulated expression of p21waf1,cip1 , a downstream component of Q13547 signalling , in A2780 ovarian carcinoma cells . R306465 showed class I HDAC isotype selectivity as evidenced by poor inhibition of Q9UBN7 ( class IIb ) confirmed by the absence of downregulation of Hsp90 chaperone c-raf protein expression and tubulin acetylation . This distinguished it from other HDAC inhibitors currently in clinical development that were either more potent towards Q9UBN7 ( e.g. vorinostat ) or had a broader HDAC inhibition spectrum ( e.g. DB06603 ) . R306465 potently inhibited cell proliferation of all main solid tumour indications , including ovarian , lung , colon , breast and prostate cancer cell lines , with IC50 values ranging from 30 to 300 nM . Haematological cell lines , including acute lymphoblastic leukaemia , acute myeloid leukaemia , chronic lymphoblastic leukaemia , chronic myeloid leukaemia , lymphoma and myeloma , were potently inhibited at a similar concentration range . R306465 induced apoptosis and inhibited angiogenesis in cell-based assays and had potent oral in vivo antitumoral activity in xenograft models . Once-daily oral administration of R306465 at well-tolerated doses inhibited the growth of A2780 ovarian , H460 lung and HCT116 colon carcinomas in immunodeficient mice . The high activity of R306465 in cell-based assays and in vivo after oral administration makes R306465 a promising novel antitumoral agent with potential applicability in a broad spectrum of human malignancies . The Conserved Kinases CDK-1 , GSK-3 , O60870 -19 , and MBK-2 Promote OMA-1 Destruction to Regulate the Oocyte-to-Embryo Transition in C. elegans . BACKGROUND : At the onset of embryogenesis , key developmental regulators called determinants are activated asymmetrically to specify the body axes and tissue layers . In C. elegans , this process is regulated in part by a conserved family of CCCH-type zinc finger proteins that specify the fates of early embryonic cells . The asymmetric localization of these and other determinants is regulated in early embryos through motor-dependent physical translocation as well as selective proteolysis . RESULTS : We show here that the CCCH-type zinc finger protein OMA-1 serves as a nexus for signals that regulate the transition from oogenesis to embryogenesis . While OMA-1 promotes oocyte maturation during meiosis , destruction of OMA-1 is needed during the first cell division for the initiation of ZIF-1-dependent proteolysis of cell-fate determinants . Mutations in four conserved protein kinase genes-mbk-2/Dyrk , kin-19/CK1alpha , gsk-3 , and cdk-1/ P06493 -cause stabilization of OMA-1 protein , and their phenotypes are partially suppressed by an oma-1 loss-of-function mutation . OMA-1 proteolysis also depends on P12004 B3 and on a ZIF-1-independent Q13617 -based E3 ubiquitin ligase complex , as well as the Q13617 -interacting protein Q7Z7L7 -11 and the Skp1-related proteins P21452 -1 and P21452 -2 . CONCLUSIONS : Our findings suggest that a P06493 / P12004 B3-dependent activity links OMA-1 proteolysis to completion of the first cell cycle and support a model in which OMA-1 functions to prevent the premature activation of cell-fate determinants until after they are asymmetrically partitioned during the first mitosis . Ontogeny of gene expression in the gonadotroph of the developing female rat . During the infantile period of the female rat ( 8-21 postnatal days [ P01160 ] of age ) , there is a dramatic increase in plasma DB00094 , which is thought to be important in initiating ovarian activity and , perhaps , the onset of puberty . To begin to understand the regulation of this DB00094 surge , we determined the ontogenetic development of LHbeta , FSHbeta , and P30968 ( P30968 ) mRNA levels in the pituitary gland throughout the infantile period of the female rat . Steady-state mRNA levels were determined by an external standard quantitative reverse transcriptase polymerase chain reaction assay . FSHbeta and P30968 mRNA levels increased to peak on P01160 12 ( p < 0.03 ) . LHbeta mRNA levels remained relatively constant until rising on P01160 18 . A DB00644 antagonist ( 10-100 microg/animal ) was administered daily from P01160 8-11 or P01160 11-13 , and animals were killed on P01160 12 or P01160 14 , respectively . FSHbeta , LHbeta , and P30968 mRNAs were not affected by DB00644 antagonist treatment . Plasma DB00094 was selectively reduced in the first group , whereas both plasma LH and DB00094 were suppressed in the second group . These data indicate that gene expression of LHbeta , FSHbeta , and P30968 are differentially regulated in the infantile female rat pituitary . DB00644 is involved in regulating the secretion of DB00094 and LH during the infantile period but not in regulating FSHbeta , LHbeta , or P30968 mRNA gene expression . P10275 inducing bladder cancer progression by promoting an epithelial-mesenchymal transition . The study investigated the role of androgen receptor ( AR ) as a potential target for the treatment of bladder cancer in regulating epithelial-mesenchymal transition or transformation ( EMT ) . Cell proliferation , and migration capacity were determined in bladder cancer T24 cells treated with small interfering RNA directed against AR , and expression levels of P12830 , β-catenin and N- cadherin were assessed using quantitative reverse transcription PCR ( qRT-PCR ) . Tumour cell growth was evaluated in vivo in T24 tumour-bearing nude mice receiving electroporation-assisted administration of anti-AR small interfering RNA . It was found that low AR expression decreased proliferation and migration of bladder cancer cells . In vivo experiments showed that silencing AR expression significantly suppressed AR-positive bladder tumour growth with decreased cell proliferation . Low AR level of T24 bladder cancer cells treated with DB01541 ( DB02901 ) decreased expression of P12830 , β-catenin and P19022 expression , indicating a strong sensitivity to the EMT and In cells with low AR content , TGF-β induced down-regulation of P12830 and β-catenin . It is concluded that suppression of AR expression decreased the production of TGF-β , inhibiting EMT and bladder cancer cell growth in vitro and in vivo , implying that its use might be a potential therapeutic target for the treatment of bladder cancer . DB01599 alleviates atherosclerosis and improves high density lipoprotein function . BACKGROUND : DB01599 is a unique hypolipidemic agent that decreases high density lipoprotein cholesterol ( HDL-C ) . However , it is not definite that whether probucol hinders the progression of atherosclerosis by improving HDL function . METHODS : Eighteen New Zealand White rabbits were randomly divided into the control , atherosclerosis and probucol groups . Control group were fed a regular diet ; the atherosclerosis group received a high fat diet , and the probucol group received the high fat diet plus probucol . Hepatocytes and peritoneal macrophages were isolated for [ (3)H ] labeled cholesterol efflux rates and expression of O95477 and SR-B1 at gene and protein levels ; venous blood was collected for serum paraoxonase 1 , myeloperoxidase activity and lipid analysis . Aorta were prepared for morphologic and immunohistochemical analysis after 12 weeks . RESULTS : Compared to the atherosclerosis group , the paraoxonase 1 activity , cholesterol efflux rates , expression of O95477 and Q8WTV0 in hepatocytes and peritoneal macrophages , and the level of O95477 and Q8WTV0 in aortic lesions were remarkably improved in the probucol group , But the serum HDL cholesterol concentration , myeloperoxidase activity , the IMT and the percentage plaque area of aorta were significantly decreased . CONCLUSION : DB01599 alleviated atherosclerosis by improving HDL function . The mechanisms include accelerating the process of reverse cholesterol transport , improving the anti-inflammatory and anti-oxidant functions . Histone deacetylase inhibitors modulate the transcriptional regulation of guanylyl cyclase/natriuretic peptide receptor-a gene : interactive roles of modified histones , histone acetyltransferase , p300 , AND Sp1 . Atrial natriuretic peptide ( P01160 ) binds guanylyl cyclase-A/natriuretic peptide receptor-A ( P16066 /NPRA ) and produces the intracellular second messenger , cGMP , which regulates cardiovascular homeostasis . We sought to determine the function of histone deacetylases ( HDACs ) in regulating Npr1 ( coding for P16066 /NPRA ) gene transcription , using primary mouse mesangial cells treated with class-specific HDAC inhibitors ( HDACi ) . Trichostatin A , a pan inhibitor , and mocetinostat ( DB05651 ) , a class I HDAC inhibitor , significantly enhanced Npr1 promoter activity ( by 8- and 10-fold , respectively ) , mRNA levels ( 4- and 5.3-fold , respectively ) , and NPRA protein ( 2.7- and 3.5-fold , respectively ) . However , MC1568 ( class II HDAC inhibitor ) had no discernible effect . Overexpression of Q13547 and Q92769 significantly attenuated Npr1 promoter activity , whereas O15379 and Q9BY41 had no effect . HDACi-treated cultured cells in vitro and intact animals in vivo showed significantly reduced binding of Q13547 and -2 and increased accumulation of acetylated H3- P35527 /14 and H4-K12 at the Npr1 promoter . Deletional analyses of the Npr1 promoter along with ectopic overexpression and inhibition of Sp1 confirmed that HDACi-induced Npr1 gene transcription is accomplished by Sp1 activation . Furthermore , HDACi attenuated the interaction of Sp1 with Q13547 /2 and promoted Sp1 association with p300 and p300/ DB02527 -binding protein-associated factor ; it also promoted the recruitment of p300 and p300/ DB02527 -binding protein-associated factor to the Npr1 promoter . Our results demonstrate that trichostatin A and DB05651 enhanced Npr1 gene expression through inhibition of Q13547 /2 and increased both acetylation of histones ( H3- P35527 /14 , H4-K12 ) and Sp1 by p300 , and their recruitment to Npr1 promoter . Our findings define a novel epigenetic regulatory mechanism that governs Npr1 gene transcription . An anti-inflammatory mechanism of taurine conjugated DB00244 against experimental colitis : taurine chloramine potentiates inhibitory effect of DB00244 on IL-1beta-mediated NFkappaB activation . Previously , we reported that oral administration of taurine conjugated DB00244 , a colon-specific prodrug of DB00244 ( DB00244 ) , is effective in ameliorating experimental colitis and taurine elicits an additive anti-inflammatory effect upon cotreatment with DB00244 . To explore a molecular mechanism for the anti-inflammatory property of the prodrug , we investigated the effect of the conjugate on IL-1beta-mediated NFkappaB activation . In human colon carcinoma Caco-2 and HCT116 cells , NFkappaB activity was accessed by a luciferase reporter assay and P05231 secretion . Protein levels were determined by Western blotting . P05231 levels were monitored by an Elisa kit . Treatment with either DB00244 or taurine chloramine ( TauCl ) inhibited IL-1beta-mediated NFkappaB dependent luciferase expression and P05231 secretion . In HCT116 cells , the inhibitory effect by TauCl or DB00244 was through preventing IL-1beta-induced O15111 activation and subsequently interfering with P25963 degradation and p65 nuclear accumulation . Furthermore , combined TauCl/ DB00244 treatment interfered additively with the activation process , leading to additive inhibitory effect on IL-1beta-mediated NFkappaB activation . Our results suggest that the anti-inflammatory effect of the prodrug on experimental colitis is attributed to the inhibition of the IL-1beta-mediated NFkappaB activation and the taurine effect is through TauCl potentiating the ability of DB00244 to inhibit IL-1beta dependent NFkappaB activation . Wnt-11 signalling controls ventricular myocardium development by patterning P19022 and beta-catenin expression . AIMS : The stage-dependent organization of the cardiomyocytes during formation of the different layers of the developing ventricular wall is critical for the establishment of a functional heart , but the instructive signals involved are still poorly known . We have addressed the potential role of Wnt-11 in the control of early ventricular myocardium assembly . METHODS AND RESULTS : We demonstrate by means of expression analysis and a mouse model in which Wnt-11 function has been inactivated that Wnt-11 is expressed by the embryonic ventricular cardiomyocytes and serves as one important signal for ventricular wall development . In the absence of Wnt-11 , the coordinated organization , intercellular contacts , co-localized expression of the cell adhesion components P19022 and beta-catenin , and the cytoskeleton of the differentiating ventricular cardiomyocytes are all disturbed . Moreover , the ventricular wall lacking Wnt-11 signalling is thinner and the expression of the Gata-4 , Nkx2.5 , Mef2c , P01160 , and DB04899 genes is down-regulated relative to controls . These defects lie behind disturbed embryonic cardiac functional development , marked by an increase in the ventricular relaxation time during the early diastole . CONCLUSION : We conclude that Wnt-11 signalling serves as a critical cell adhesion cue for the organization of the cardiomyocytes in the developing ventricular wall , which is essential for the establishment of a functional heart . Effect of matrine on the expression of DB05875 receptor and inflammatory cytokines production in human skin keratinocytes and fibroblasts . Matrine is a kind of alkaloid found in certain Sophora plants , which has been extensively used in China for the treatment of viral hepatitis , cancer , cardiac diseases and skin diseases ( such as atopic dermatitis and eczema ) . It also has been confirmed that DB05875 ( SP ) and its receptor ( neurokinin-1 receptor , P25103 ) are involved in the pathogenesis of inflammatory skin disorders . So the present study was designed to investigate the effect of matrine on the expression of P25103 and cytokines production induced by SP in HaCaT cells ( a human epidermal keratinocyte cell line ) and dermal fibroblasts . In addition , cell viability was also evaluated . The results showed that matrine inhibited the expression of P25103 in HaCaT cells and fibroblasts . SP induced the production of interleukin ( IL ) -1beta , P10145 , interferon ( IFN ) -gamma , and monocyte chemotactic protein ( MCP ) -1 in both cell types . Matrine 5-100 microg/mL had little effect on cell viability . It inhibited SP-induced IL-1beta , P10145 and P13500 production in HaCaT cells and fibroblasts , while it increased the production of P01579 in HaCaT cells . Both SP and matrine had no effect on the secretion of P05231 . These findings suggest that matrine may have potential treatment function on SP related cutaneous inflammation by inhibition of the expression of DB05875 receptor and regulation of the production of inflammatory cytokines .

MH_train_1312

MH_train_1312

MH_train_1312

interacts_with DB08820?

multiple_choice

DB08888 for vitreoretinal diseases . P02751 and laminin are clinically relevant plasmin receptors in the eye . Located at the vitreoretinal interface , they are cleaved by ocriplasmin ( DB05028 , ThromboGenics , Iselin , NJ ) , a novel ophthalmic medication . A series of clinical trials to study ocriplasmin for the treatment of vitreoretinal diseases such as vitreomacular traction , macular hole , and exudative age-related macular degeneration are underway . The results are promising and may impact patient care . DB09280 - DB08820 in Patients with Cystic Fibrosis Homozygous for Phe508del P13569 . DB08820 potentiation of multiple P13569 channels with gating mutations . BACKGROUND : The investigational P13569 potentiator ivacaftor ( VX-770 ) increased P13569 channel activity and improved lung function in subjects with CF who have the G551D P13569 gating mutation . The aim of this in vitro study was to determine whether ivacaftor potentiates mutant P13569 with gating defects caused by other P13569 gating mutations . METHODS : The effects of ivacaftor on P13569 channel open probability and chloride transport were tested in electrophysiological studies using Fischer rat thyroid ( P17948 ) cells expressing different P13569 gating mutations . RESULTS : DB08820 potentiated multiple mutant P13569 forms with defects in P13569 channel gating . These included the G551D , G178R , S549N , S549R , G551S , G970R , G1244E , S1251N , S1255P and G1349D P13569 gating mutations . CONCLUSION : These in vitro data suggest that ivacaftor has a similar effect on all P13569 forms with gating defects and support investigation of the potential clinical benefit of ivacaftor in CF patients who have P13569 gating mutations beyond G551D . A phase-1 trial of bexarotene and denileukin diftitox in patients with relapsed or refractory cutaneous T-cell lymphoma . DB00004 , a genetically engineered fusion protein combining the enzymatically active domains of diphtheria toxin and the full-length sequence for interleukin-2 ( P60568 ) , efficiently targets lymphoma cells expressing the high-affinity P60568 receptor ( IL-2R ) consisting of the alpha/p55/CD25 , beta/p75/CD122 , and gamma/ P31785 /CD132 chains . In vitro studies demonstrated that the retinoid X receptor ( RXR ) retinoid , bexarotene , at biologically relevant concentrations of 10(-6) M to 10(-8) M , upregulated both the p55 and p75 subunits of the IL-2R and enhanced 5- to 10-fold the susceptibility of T-cell leukemia cells to denileukin diftitox . To determine whether this biomodulatory effect could be recapitulated in vivo , we treated 14 patients with relapsed or refractory cutaneous T-cell lymphoma with escalating doses of bexarotene ( 75 mg/day-300 mg/day ) and denileukin diftitox ( 18 mcg/kg per day x 3 days every 21 days ) in a phase 1 trial . Overall response was 67 % ( 4 complete responses , 4 partial responses ) . Modulation of IL-2R expression was observed at or above a bexarotene dose of 150 mg/day . Four patients experienced grade 2 or 3 leukopenia , and 2 had grade 4 lymphopenia . Our results demonstrate that the combination of denileukin diftitox and bexarotene is well tolerated and that even low doses ( 150 mg/day ) of bexarotene are capable of in vivo upregulation of CD25 expression on circulating leukemia cells . Multiple arrhythmic syndromes in a newborn , owing to a novel mutation in Q14524 . BACKGROUND : Mutations in the Q14524 gene have been linked to Brugada syndrome ( BrS ) , conduction disease , Long QT syndrome ( LQT3 ) , atrial fibrillation ( AF ) , and to pre- and neonatal ventricular arrhythmias . OBJECTIVE : The objective of this study is to characterize a novel mutation in Na(v)1.5 found in a newborn with fetal chaotic atrial tachycardia , post-partum intraventricular conduction delay , and QT interval prolongation . METHODS : Genomic DNA was isolated and all exons and intron borders of 15 ion-channel genes were sequenced , revealing a novel missense mutation ( Q270K ) in Q14524 . Na(v)1.5 wild type ( WT ) and Q270K were expressed in CHO- P04264 with and without the Na(v)β1 subunit . Results . Patch-clamp analysis showed ∼40 % reduction in peak sodium channel current ( I(Na) ) density for Q270K compared with WT . Fast and slow decay of I(Na) were significantly slower in Q270K . Steady-state activation and inactivation of Q270K channels were shifted to positive potentials , and window current was increased . The tetrodotoxin-sensitive late I(Na) was increased almost 3-fold compared with WT channels . DB00243 reduced late I(Na) in WT and Q270K channels , while exerting minimal effects on peak I(Na) . CONCLUSION : The Q270K mutation in Q14524 reduces peak I(Na) while augmenting late I(Na) , and may thus underlie the development of atrial tachycardia , intraventricular conduction delay , and QT interval prolongation in an infant . Coinduction of endothelial nitric oxide synthase and arginine recycling enzymes in aorta of diabetic rats . Decreased availability of arginine and impaired production of NO ( nitric oxide ) have been implicated in the development of endothelial dysfunction . DB00155 formed by the NOS reaction is recycled to arginine by the citrulline-NO cycle , which is composed of NOS , argininosuccinate synthetase ( AS ) , and argininosuccinate lyase . Therefore , we investigated the alterations of these enzymes in the aorta of streptozotocin ( Q11206 ) -induced diabetic rats . P29474 and AS mRNAs were increased by three- to fourfold 1-2 weeks after Q11206 treatment and decreased at 4 weeks . AL mRNA was weakly induced . Induction of P29474 and AS proteins was also observed . Cationic amino acid transporter ( CAT ) -1 mRNA remained little changed , and CAT-2 mRNA was not detected . The plasma nitrogen oxide levels were increased 1-2 weeks after Q11206 treatment and decreased at 4 weeks . Transforming growth factor-beta1 ( TGF-beta1 ) mRNA in the aorta was also induced . TGF-beta1 induced P29474 and AS mRNAs in human umbilical vein endothelial cells but inhibited the proliferation of HUVEC . These results indicate that P29474 and AS are coinduced in the aorta in early stages of Q11206 -induced diabetic rats and that the induction is mediated by TGF-beta1 . The results also suggest that TGF-beta1 works antiatherogenically at early stages of diabetes by increasing NO production , whereas prolonged elevation of TGF-beta1 functions atherogenically by inhibiting endothelial cell growth . Managing the underlying cause of cystic fibrosis : a future role for potentiators and correctors . Cystic fibrosis ( CF ) , a severe genetic disease , is caused by mutations that alter the structure and function of P13569 , a plasma membrane channel permeable to chloride and bicarbonate . Defective anion transport in CF irreversibly damages the lungs , pancreas , liver , and other organs . CF mutations cause loss of P13569 function in multiple ways . In particular , class 3 mutations such as p.Gly551Asp strongly decrease the time spent by P13569 in the open state ( gating defect ) . Instead , class 2 mutations impair the maturation of P13569 protein and its transport from the endoplasmic reticulum to the plasma membrane ( trafficking defect ) . The deletion of phenylalanine 508 ( p.Phe508del ) , the most frequent mutation among CF patients ( 70-90 % ) , destabilizes the P13569 protein , thus causing both a trafficking and a gating defect . These two defects can be overcome with drug-like molecules generically called correctors and potentiators , respectively . The potentiator Kalydeco™ ( also known as DB08820 or VX-770 ) , developed by Vertex Pharmaceuticals , has been recently approved by the US FDA and the European Medicines Agency ( P15941 ) for the treatment of CF patients carrying at least one P13569 allele with the p.Gly551Asp mutation ( 2-5 % of all patients ) . In contrast , the corrector VX-809 , which significantly improves p.Phe508del- P13569 trafficking in vitro , is still under study in clinical trials . Because of multiple defects caused by the p.Phe508del mutation , it is probable that rescue of the mutant protein will require combined treatment with correctors having different mechanisms of action . This review evaluates the status of experimental and clinical research in pharmacotherapy for the CF basic defect . Phosphorylation and recycling kinetics of G protein-coupled receptors . The rate of ligand-induced phosphorylation of the V2 and V1a vasopressin receptors was characterized in P29320 293 cells . Both receptors were phosphorylated predominantly by GRKs , and the V1a receptor was also phosphorylated by protein kinase C regardless of the presence or absence of ligand . Phosphorylation of the P37288 catalyzed by GRKs reached maximal values at the shortest measured time : 15 seconds , and decayed rapidly with a t1/2 of 6 min in the continuous presence of AVP . In agreement with the hypothesis that dephosphorylation must precede receptor recycling to the cell surface , the P37288 returned rapidly to the cell surface after removal of the hormone from the medium . Phosphate incorporation into the P30518 proceeded at a slower pace , and the internalized phosphorylated receptor failed to recycle to the cell surface and retained its phosphate for a long time in the presence or absence of ligand . A single mutation in the carboxy terminus of the P30518 accelerated de-phosphorylation of the protein and conferred recycling properties to the P30518 . These experiments provided molecular evidence for the hypothesis that internalization is required for de-phosphorylation and recycling of reactivated G protein coupled receptors to the cell surface . Preeclampsia : a bioinformatics approach through protein-protein interaction networks analysis . BACKGROUND : In this study we explored preeclampsia through a bioinformatics approach . We create a comprehensive genes/proteins dataset by the analysis of both public proteomic data and text mining of public scientific literature . From this dataset the associated protein-protein interaction network has been obtained . Several indexes of centrality have been explored for hubs detection as well as the enrichment statistical analysis of metabolic pathway and disease . RESULTS : We confirmed the well known relationship between preeclampsia and cardiovascular diseases but also identified statistically significant relationships with respect to cancer and aging . Moreover , significant metabolic pathways such as apoptosis , cancer and cytokine-cytokine receptor interaction have also been identified by enrichment analysis . We obtained P17948 , P15692 , P02751 , F2 and P49763 genes with the highest scores by hubs analysis ; however , we also found other genes as P30101 , P07948 , O14492 and Q92597 with high scores . CONCLUSIONS : The applied methodology not only led to the identification of well known genes related to preeclampsia but also to propose new candidates poorly explored or completely unknown in the pathogenesis of preeclampsia , which eventually need to be validated experimentally . Moreover , new possible connections were detected between preeclampsia and other diseases that could open new areas of research . More must be done in this area to resolve the identification of unknown interactions of proteins/genes and also for a better integration of metabolic pathways and diseases . Role of calpain-1 in the early phase of experimental P35858 . Elevation in [Ca(2+)]i and activation of calpain-1 occur in central nervous system of P00441 (G93A) transgenic mice model of amyotrophic lateral sclerosis ( P35858 ) , but few data are available about the early stage of P35858 . We here investigated the level of activation of the Ca(2+)-dependent protease calpain-1 in spinal cord of P00441 (G93A) mice to ascertain a possible role of the protease in the aetiology of P35858 . Comparing the events occurring in the 120 day old mice , we found that [Ca(2+)]i and activation of calpain-1 were also increased in the spinal cord of 30 day old mice , as indicated by the digestion of some substrates of the protease such as P29475 , αII-spectrin , and the Q13224 subunit of DB01221 -R . However , the digestion pattern of these proteins suggests that calpain-1 may play different roles depending on the phase of P35858 . In fact , in spinal cord of 30 day old mice , activation of calpain-1 produces high amounts of P29475 active species , while in 120 day old mice enhanced-prolonged activation of calpain-1 inactivates P29475 and down-regulates Q13224 . Our data reveal a critical role of calpain-1 in the early phase and during progression of P35858 , suggesting new therapeutic approaches to counteract its onset and fatal course . P13569 mutations impart elevated immune reactivity in a murine model of cystic fibrosis related diabetes . Increased life expectancy in cystic fibrosis ( CF ) is accompanied by an increasing incidence of CF related diabetes ( CFRD ) . Altered immune reactivity occurs in CF , which we hypothesize , is exacerbated by hyperglycemia . P13569 deficient ( P13569 -/- ) mice were rendered hyperglycemic by streptozotocin ( Q11206 ) to test this hypothesis . P13569 -/- , C57BL/6J , and FVB/NJ mice received either Q11206 or lactated ringers ( LR ) ( n=5-10 ) . Four weeks later , splenocytes were harvested , mitogen stimulated , and analyzed for cytokine production ( P60568 , P05112 , and P22301 ) along with stimulation indices ( SI ) . SI of Q11206 -treated P13569 -/- were elevated compared to LR-treated mice , although both were greater than C57BL/6J and FVB/NJ ( p < 0.05 ) . Fasting glucose levels of Q11206 -treated P13569 -/- mice correlated with SI ( p < 0.003 ) . Stimulated P22301 concentrations were elevated in Q11206 -treated P13569 -/- compared to LR-treated animals and controls ( p < 0.05 ) . P60568 levels were greater in P13569 -/- mice compared to controls ( p < 0.05 ) , but unrelated to Q11206 . Reinforcing generalized cytokine up-regulation in P13569 -/- , P05112 levels were greater in P13569 -/- mice compared to C57BL/6J , but FVB/NJ mice demonstrated greatest concentrations following Q11206 . These results suggest that , hyperglycemia may exacerbate the clinical course in CF by impacting immune reactivity . There is clear need to maximize metabolic management in CFRD . Inhibition of the liver expression of arylalkylamine N-acetyltransferase increases the expression of angiogenic factors in cholangiocytes . BACKGROUND AND AIMS : Reduction of biliary serotonin N-acetyltransferase ( Q16613 ) expression and melatonin administration/secretion in cholangiocytes increases biliary proliferation and the expression of SR , P13569 and Cl(-)/HCO3 ( - ) P04920 . The balance between biliary proliferation/damage is regulated by several autocrine neuroendocrine factors including vascular endothelial growth factor-A/C ( P15692 /C ) . VEGFs are secreted by several epithelia , where they modulate cell growth by autocrine and paracrine mechanisms . No data exists regarding the effect of Q16613 modulation on the expressions of VEGFs by cholangiocytes . METHODS : In this study , we evaluated the effect of local modulation of biliary Q16613 expression on the cholangiocytes synthesis of P15692 /C . RESULTS : The decrease in Q16613 expression and subsequent lower melatonin secretion by cholangiocytes was associated with increased expression of P15692 /C . Overexpression of Q16613 in cholangiocyte lines decreased the expression of P15692 /C . CONCLUSIONS : Modulation of melatonin synthesis may affect the expression of P15692 /C by cholangiocytes and may modulate the hepatic microvascularization through the regulation of P15692 /C expression regulating biliary functions . Targeting autocrine and paracrine P15692 receptor pathways inhibits human lymphoma xenografts in vivo . The role of angiogenesis in lymphoproliferative diseases is not well established . We demonstrate here that human lymphoma cells secrete vascular endothelial growth factor ( P15692 ) and express P15692 receptor 1 ( P17948 ) and P35968 . Proliferation of non-Hodgkin lymphoma ( Q9NZ71 ) cells under serum-free conditions was enhanced by the addition of P15692 and was blocked by P17948 - and P35968 -specific antibodies . To differentiate between P15692 -mediated autocrine and paracrine effects on lymphoma growth , NOD/SCID mice engrafted with human diffuse large B-cell lymphoma ( DLBCL ) were treated with species-specific antibodies against human P17948 ( 6.12 ) , human P35968 ( DB06101 ) , murine P17948 ( MF-1 ) , or murine P35968 ( DC101 ) . Treatment with 6.12 or DC101 ( targeting tumor P17948 and host P35968 ) reduced established DLBCL xenograft growth , whereas treatment with DB06101 or MF-1 ( targeting tumor P17948 and host P17948 ) had no effect . Decreased tumor volumes after 6.12 and DC101 treatment correlated with increased tumor apoptosis and reduced vascularization , respectively , supporting the presence of autocrine P17948 - and paracrine P35968 -mediated pathways in lymphomagenesis . Inhibition of paracrine P15692 interactions ( DC101 ) in these models was equivalent to their inhibition with rituximab . Combining DC101 with therapeutic agents ( rituximab , 6.12 , methotrexate ) consistently improved tumor responses over those of single-agent therapy . These data support the further clinical development of VEGFR-targeted approaches for the therapy of aggressive DLBCL . Reduction of neuronal and inducible nitric oxide synthase gene expression in patients with cystic fibrosis . As a consequence of diminished nitric oxide synthase ( NOS ) protein concentration , the airway concentration of nitric oxide ( NO ) is reduced in patients with cystic fibrosis ( CF ) . This appears to lead to a reduced elimination of such microorganisms as Pseudomonas aeruginosa . The objective of this study was to analyze whether inducible ( P35228 ) , endothelial ( P29474 ) and neuronal ( P29475 ) NOS are reduced at mRNA level and if so whether this is caused directly by the defective CF transmembrane conductance regulator ( P13569 ) . Nasal polyps from three patients with CF and four otherwise healthy patients were obtained . The expression of the three NOS isoenzymes was quantified using real-time PCR . The P35228 expression was assessed in colon carcinoma cells ( CaCo ) transfected with a normal and a mutated ( DeltaF508 ) P13569 . In CF patients , P35228 mRNA expression was 10-to 20-fold and P29475 gene expression was one-fifth to one-tenth that in control patients ( P < 0.001 ) . In CaCo cells , P35228 gene expression under basal and endotoxin-stimulated conditions did not differ between cells transfected with a mutated P13569 and those transfected with an intact P13569 . This observation suggests that cystic fibrosis is associated with reduced P35228 and P29475 gene expression in nasopharyngeal tissue , possibly disturbing the barrier against infective agents already at the site of entrance . P62158 -mediated effects of loperamide on chloride transport by brush border membrane vesicles from human ileum . We investigated whether the synthetic opiate loperamide-HCl is able to regulate specific transport systems for sodium and chloride in brush border membrane vesicles ( BBMVs ) from human ileum and whether such activities are mediated by calcium/calmodulin . In BBMVs we studied Na+/H+ antiport , Cl+/OH- antiport , Na+/Cl- cotransport , and the Cl- conductive pathway . Brush border membrane vesicles were incubated with 10 microM loperamide over 4 h at 5 degrees C before the uptake experiments . In ileal BBMVs , loperamide stimulated intravesicular accumulation of Na+ in the presence of Cl- and vice versa . After 1 min of incubation , the stimulatory effect was 35 % +/- 5 % ( p less than 0.005 ) of the control without loperamide . DB00836 also stimulated Cl-/OH- antiport by 30 % +/- 5 % ( p less than 0.005 ) in BBMVs of ileum . In addition , we studied the role of Ca2+/calmodulin in the action of loperamide on chloride transport by human BBMVs . In loperamide-pretreated BBMVs , calmodulin activity was significantly decreased ( 12 +/- 2 vs. 38 +/- 4 pmol/mg protein ) . When loperamide-pretreated vesicles were incubated with 2 microM calcium ( free concentration ) plus 5 microM calmodulin for 1 h at 5 degrees C , complete inhibition of the stimulatory effect of loperamide on Cl-/OH- antiport and Na+/Cl- cotransport was observed . Increasing the Ca2+/calmodulin activity of loperamide-pretreated BBMVs with 2 microM calcium plus 5 microM calmodulin led to a significant inhibition of Cl-/OH- antiport and Na+/Cl- cotransport by 40 % +/- 5 % ( p less than 0.005 ) . Nutritional correlates of acute respiratory infections . During October 1992 to June 1993 in eight villages covered by the Primary Health Center Machhra in Meerut District , India , interviews with mothers and examinations of 1600 children aged less than 5 years ( under-fives ) were conducted to examine the relationship between acute respiratory infection ( Q9Y4X5 ) and malnutrition . 42.25 % of all children had an Q9Y4X5 within the last 15 days . Most ARIs ( 73.4 % ) were considered mild ( cough and cold with no pneumonia ) . Pneumonia accounted for 19.5 % of all Q9Y4X5 cases , which were considered moderate . The remaining Q9Y4X5 cases were severe ( severe and very severe pneumonia ) . 57.5 % of all children suffered from protein energy malnutrition ( P15941 ) . 78.6 % of children aged 12-14 months had P15941 . Q9Y4X5 was more common among malnourished children than well-nourished children ( 52.2 % vs. 28.8 % ; p 0.001 ) . The incidence of Q9Y4X5 increased as the nutritional status deteriorated ( p 0.05 ) . It also increased as the midarm circumference decreased ( p 0.001 ) . These findings confirm the synergistic action between malnutrition and infection , in this case Q9Y4X5 . Malnourished children suffer considerable impairment in immunity , especially cellular immunity , which makes them more prone to Q9Y4X5 . These findings reinforce the need to strengthen the quality , quantity , and accessibility of nutritional services , particularly promotion of breast feeding and vitamin A supplementation . P13569 modulates neurosecretory function in pulmonary neuroendocrine cell-related tumor cell line models . The pulmonary neuroendocrine cell ( PNEC ) system consists of solitary cells and distinctive cell clusters termed neuroepithelial bodies ( P20929 ) localized in the airway epithelium . PNEC/ P20929 express a variety of bioactive substances , including amine ( serotonin , 5HT ) and neuropeptides . We have previously shown that P20929 cells are O(2) sensors expressing nicotinamide adenine diphosphate oxidase complex and O(2) sensitive K(+) channel . Recently , we demonstrated expression of functional cystic fibrosis transmembrane conductance regulator ( P13569 ) and Cl(-) conductances in P20929 cells of rabbit neonatal lung . Because PNEC/ P20929 are sparsely distributed and difficult to study in native lung , we investigated small-cell lung carcinoma ( SCLC ) and carcinoid tumor cell lines ( tumor counterparts of normal PNEC/ P20929 ) as models for PNEC/ P20929 . SCLC ( H146 , H345 ) and carcinoid ( H727 ) cell lines express neuroendocrine cell markers , including chromogranin A , neural cell adhesion molecule ( N- P62158 ) , 5HT , and tryptophan hydroxylase . We report that H146 , H345 , and H727 express P13569 messenger RNA ( reverse transcription polymerase chain reaction ) and protein ( immunoblotting ) and possess functional P13569 Cl(-) conductance , demonstrated by an iodide efflux assay inhibitable by transfection with antisense P13569 . Using an immunoassay to quantitate 5HT secretion , we also show that downregulation of P13569 abolishes hypoxia-induced 5HT release , and reduces secretory response to high potassium . Our findings suggest that P13569 may modulate neurosecretory activity of PNEC/ P20929 possessing O(2) sensor function . We propose that these tumor cell lines may be useful models for investigating the role of P13569 in PNEC/ P20929 functions in health and disease . Effects of neurosteroids on Ca(2+) signaling mediated by recombinant N-methyl-D-aspartate receptor expressed in Chinese hamster ovary cells . DB02789 sulfate ( PREGS ) potentiates the N-methyl-D-aspartate ( DB01221 ) receptor-mediated Ca(2+)-signals in cultured hippocampal neurons . The DB01221 receptor family has several different subunits whose expression in neurons has distinct spatial and temporal patterns . To examine subunit specificity of the PREGS action , we have investigated the effect of PREGS on recombinant GluR epsilon2/zeta1 ( Q13224 / Q9UHB4 ) type DB01221 receptors stabley expressed in Chinese hamster ovary cells using heat shock promoters . PREGS enhanced the Ca(2+) influx through the GluR epsilon2/zeta1 receptors in a dose-dependent manner . The EC(50) of PREGS for the GluR epsilon2/zeta1 receptors was 8.6 microM . Other sulfated neurosteroids , DB05804 ( DHEAS ) , 17beta-estradiol sulfate and 3alpha-ol-5beta-pregnan-20-one sulfate ( 3alpha5betaS ) , inhibited the positive modulatory effect of PREGS on the GluR epsilon2/zeta1 DB01221 receptors . DB08954 , a specific inhibitor of GluR epsilon2 subunit , abolished the DB01221 -induced Ca(2+) influx even in the presence of PREGS . These results imply that PREGS positively modulates the Ca(2+) influx through the GluR epsilon2/zeta1 receptors which are expressed from the embryonic period . The testis anion transporter TAT1 ( Q96RN1 ) physically and functionally interacts with the cystic fibrosis transmembrane conductance regulator channel : a potential role during sperm capacitation . The Slc26 gene family encodes several conserved anion transporters implicated in human genetic disorders , including Pendred syndrome , diastrophic dysplasia and congenital chloride diarrhea . We previously characterized the TAT1 ( testis anion transporter 1 ; Q96RN1 ) protein specifically expressed in male germ cells and mature sperm and showed that in the mouse , deletion of Tat1 caused male sterility due to a lack of sperm motility , impaired sperm capacitation and structural defects of the flagella . Ca(2+) , Cl(-) and HCO(3)(-) influxes trigger sperm capacitation events required for oocyte fertilization ; these events include the intracellular rise of cyclic adenosine monophosphate ( DB02527 ) and protein kinase A ( PKA ) -dependent protein phosphorylation . The cystic fibrosis transmembrane conductance regulator ( P13569 ) is expressed in mature sperm and has been shown to contribute to Cl(-) and HCO(3)(-) movements during capacitation . Furthermore , several members of the SLC26 family have been described to form complexes with P13569 , resulting in the reciprocal regulation of their activities . We show here that TAT1 and P13569 physically interact and that in Xenopus laevis oocytes and in CHO- P04264 cells , TAT1 expression strongly stimulates P13569 activity . Consistent with this , we show that Tat1 inactivation in mouse sperm results in deregulation of the intracellular DB02527 content , preventing the activation of PKA-dependent downstream phosphorylation cascades essential for sperm activation . These various results suggest that TAT1 and P13569 may form a molecular complex involved in the regulation of Cl(-) and HCO(3)(-) fluxes during sperm capacitation . In humans , mutations in P13569 and/or TAT1 may therefore be causes of asthenozoospermia and low fertilizing capacity of sperm . Mechanism of AVP release and synthesis in chronic salt-loaded rats . DB00067 ( AVP ) is involved in osmotic regulation in the brain and peripheral tissues . To elucidate the regulatory mechanism that involves AVP release in hyperosmolality , we investigated the regulation of the synthesis and release of AVP in chronic salt-loaded rats . In chronic salt-loaded rats , which were generated by free access to water containing 2 % NaCl for 7 days , plasma osmolality was significantly increased compared with control value . When tested , the AVP content was significantly higher in plasma but lower in the pituitary and whole brain ( hypothalamus , cortex and striatum ) than in control rats . The expression of AVP mRNA in the brain was significantly up-regulated compared with that in control rats . These data lead to the suggestion that hyperosmolality stimulates AVP release from the brain and subsequently induces AVP synthesis in the brain . On the other hand , mRNA levels of vasopressin V1a receptor ( P37288 ) , whose down-regulation is known to be a counteraction to the P37288 activation , was not changed in the brain , suggesting that the AVP seems not to interact with the P37288 in the brain . These results suggest that hyperosmosis promotes the release of AVP into plasma , the subsequent induction of AVP mRNA in the brain and its action on the peripheral tissues . Drosophila Answers to Q13148 Proteinopathies . Initially implicated in the pathogenesis of P13569 and HIV-1 transcription , nuclear factor Q13148 was subsequently found to be involved in the origin and development of several neurodegenerative diseases . In 2006 , in fact , it was reported for the first time the cytoplasmic accumulation of Q13148 in ubiquitin-positive inclusions of P35858 and FTLD patients , suggesting the presence of a shared underlying mechanism for these diseases . Today , different animal models of Q13148 proteinopathies are available in rodents , nematodes , fishes , and flies . Although these models recapitulate several of the pathological features found in patients , the mechanisms underpinning the progressive neuronal loss observed in Q13148 proteinopathies remain to be characterized . Compared to other models , Drosophila are appealing because they combine the presence of a sophisticated brain with the possibility to investigate quickly and massively phenotypic genetic modifiers as well as possible therapeutic strategies . At present , the development of Q13148 -related Drosophila models has further strengthened the hypothesis that both Q13148 " loss-of-function " and " gain-of-function " mechanisms can contribute to disease . The aim of this paper is to describe and compare the results obtained in a series of transgenic and knockout flies , along with the information they have generated , towards a better understanding of the mechanisms underlying Q13148 proteinopathies . Interplay between inhibitory ferric and stimulatory curcumin regulates phosphorylation-dependent human cystic fibrosis transmembrane conductance regulator and ΔF508 activity . Curcumin potentiates cystic fibrosis transmembrane conductance regulator ( P13569 ) activation in an DB00171 -independent but phosphorylation-dependent manner . The underlying molecular mechanisms are unclear . Here , P29320 -293T cells cultured in an Fe(3+)-containing medium were transiently transfected with P13569 constructs , and the role of the inhibitory Fe(3+) bridge between intracellular loop 3 and the regulatory domain of P13569 in this pathway was investigated . The results showed that ethylenediaminetetraacetic acid ( DB00974 ) stimulated phosphorylation-dependent P13569 activation and the stimulation was suppressed by the deletion of the regulatory domain or the insertion of a C832A mutation that removes the Fe(3+)-binding interface . Furthermore , curcumin potentiation of P13569 was significantly weakened not only by Fe(3+)-insensitive mutations at the interface between the regulatory domain and intracellular loop 3 but also by N-ethylmaleimide or DB00974 pretreatment that removes Fe(3+) . More importantly , potentiation of P13569 was completely suppressed by sufficient Fe(3+) . Finally , the insertion of Fe(3+)-insensitive H950R/S768R increased the curcumin-independent activity of ΔF508 but weakened its curcumin potentiation . Thus , Fe(3+) homeostasis in epithelia may play a critical role in regulating P13569 activity , and targeting Fe(3+)-chelating potentiators may direct new therapies for cystic fibrosis . Selective inhibition of the tumor marker O60218 by antiinflammatory N-phenylanthranilic acids and glycyrrhetic acid . A human aldose reductase-like protein , O60218 in the aldo-keto reductase ( AKR ) superfamily , was recently identified as a tumor marker of several types of cancer . DB02383 , an aldose reductase inhibitor ( Q9Y4X5 ) , is known to be the most potent inhibitor of the enzyme . In this study , we compared the inhibitory effects of other ARIs including flavonoids on O60218 and aldose reductase to evaluate their specificity . However , ARIs showed lower inhibitory potency for O60218 than for aldose reductase . In the search for potent and selective inhibitors of O60218 from other drugs used clinically , we found that non-steroidal antiinflammatory N-phenylanthranilic acids , diclofenac and glycyrrhetic acid competitively inhibited O60218 , showing K(i) values of 0.35-2.9 microM and high selectivity to this enzyme ( 43-57 fold versus aldose reductase ) . Molecular docking studies of mefenamic acid and glycyrrhetic acid in the O60218 -nicotinamide adenine dinucleotide phosphate ( NADP(+) ) complex and site-directed mutagenesis of the putative binding residues suggest that the side chain of Val301 and a hydrogen-bonding network among residues Val301 , Gln114 and Ser304 are important for determining the inhibitory potency and selectivity of the non-steroidal antiinflammatory drugs . Thus , the potent and selective inhibition may be related to the cancer chemopreventive roles of the drugs , and their structural features may facilitate the design of new anti-cancer agents targeting O60218 .

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interacts_with DB01267?

multiple_choice

Short-term biomarker modulation prevention study of anastrozole in women at increased risk for second primary breast cancer . The selective estrogen receptor modulators ( SERM ) , Tamoxifen and raloxifen reduce risk breast cancer . Patient acceptance of SERMs for breast cancer prevention is low due to toxicities . New agents with a better toxicity profile are needed . P11511 inhibitors ( AI ) reduce the risk of contralateral breast cancer and risk of new breast cancer in high risk women . However , the mechanism by which AIs reduce breast risk is not known . Surrogate biomarkers are needed to evaluate the effect of preventive agents . The objective of this prospective short-term prevention study was to evaluate the effect of anastrozole on biomarkers in breast tissue and serum of women at increased risk for developing a contralateral breast cancer . Women with a history of stage I , II breast cancer who started anastrozole for standard adjuvant treatment were eligible . Patients underwent baseline fine needle aspiration of the unaffected breast and serum collection for biomarker analysis before starting anastrozole at 1 mg per oral/day and again at 6 months . Biomarkers included changes in cytology , insulin-like growth factor 1 ( DB01277 ) , P08833 ( P08833 ) , and P17936 . Thirty-seven patients were enrolled . There was a significant modulation in serum P08833 levels between pre- and postsamples ( P = 0.02 ) . No change was observed in DB01277 , P17936 , and breast cytology.We showed a significant modulation of P08833 levels with six months anastrozole . DB01217 is currently being studied as a prevention agent in a large phase III trial and our results provide support for continued evaluation of P08833 as a surrogate endpoint biomarker in prospective breast chemoprevention studies . P08571 dependence of O00206 endocytosis and Q8IUC6 signaling displays ligand specificity and is dissociable in endotoxin tolerance . Dimerization of O00206 ( O00206 ) /myeloid differentiation factor 2 ( Q9Y6Y9 ) heterodimers is critical for both MyD88- and TIR-domain-containing adapter-inducing IFN-β ( Q8IUC6 ) -mediated signaling pathways . Recently , Zanoni et al. [ ( 2011 ) Cell 147(4):868-880 ] reported that cluster of differentiation 14 ( P08571 ) is required for LPS-/Escherichia coli- induced O00206 internalization into endosomes and activation of Q8IUC6 -mediated signaling in macrophages . We confirmed their findings with LPS but report here that P08571 is not required for receptor endocytosis and downstream signaling mediated by O00206 / Q9Y6Y9 agonistic antibody ( UT12 ) and synthetic small-molecule O00206 ligands ( 1Z105 ) in murine macrophages . P08571 deficiency completely ablated the LPS-induced Q9UHD2 / Q14653 signaling axis that mediates production of IFN-β in murine macrophages without affecting MyD88-mediated signaling , including NF-κB , MAPK activation , and P01375 -α and P05231 production . However , neither the MyD88- nor Q8IUC6 -signaling pathways and their associated cytokine profiles were altered in the absence of P08571 in UT12- or 1Z105-treated murine macrophages . DB04933 ( E5564 ) , a lipid A antagonist that binds the Q9Y6Y9 " pocket , " completely blocked LPS- and 1Z105-driven , but not UT12-induced , O00206 dimerization and endocytosis . Furthermore , O00206 endocytosis is induced in macrophages tolerized by exposure to either LPS or UT12 and is independent of P08571 . These data indicate that O00206 receptor endocytosis and the Q8IUC6 -signaling pathway are dissociable and that O00206 internalization in macrophages can be induced by UT12 , 1Z105 , and during endotoxin tolerance in the absence of P08571 . Update and new concepts in vitamin responsive disorders of folate transport and metabolism . Derivatives of folic acid are involved in transfer of one-carbon units in cellular metabolism , playing a role in synthesis of purines and thymidylate and in the remethylation of homocysteine to form methionine . Five inborn errors affecting folate transport and metabolism have been well studied : hereditary folate malabsorption , caused by mutations in the gene encoding the proton-coupled folate transporter ( Q96NT5 ) ; glutamate formiminotransferase deficiency , caused by mutations in the O95954 gene ; methylenetetrahydrofolate reductase deficiency , caused by mutations in the P42898 gene ; and functional methionine synthase deficiency , either as the result of mutations affecting methionine synthase itself ( cblG , caused by mutations in the Q99707 gene ) or affecting the accessory protein methionine synthase reductase ( cblE , caused by mutations in the Q9UBK8 gene ) . Recently additional inborn errors have been identified . Cerebral folate deficiency is a clinically heterogeneous disorder , which in a few families is caused by mutations in the P15328 gene . P00374 deficiency is characterized by megaloblastic anemia and cerebral folate deficiency , with variable neurological findings . It is caused by mutations in the P00374 gene . Deficiency in the trifunctional enzyme containing methylenetetrahydrofolate dehydrogenase , methenyltetrahydrofolate cyclohydrolase and formyltetrahydrofolate synthetase activities , has been identified in a single patient with megaloblastic anemia , atypical hemolytic uremic syndrome and severe combined immune deficiency . It is caused by mutations in the P11586 gene . P01583 ( P01583 ) and tumor necrosis factor alpha ( P01375 alpha ) regulate insulin-like growth factor binding protein-1 ( P08833 ) levels and mRNA abundance in vivo and in vitro . P01375 alpha and P01583 are thought to contribute to impaired anabolism in a variety of clinical states , including sepsis , cancer cachexia and the AIDS wasting syndrome . We asked whether cytokines exert direct effects on hepatic production of P08833 , an important modulator of IGF bioavailability . C57BL/6 mice were treated with 100 micrograms/kg of recombinant P01583 or P01375 alpha by intraperitoneal injection . Western ligand blotting and immunoprecipitation with specific antisera revealed that serum levels of P08833 ( but not P18065 , -3 , -4 , -5 or -6 ) are increased approximately 4 fold 2 h after treatment and then decline . Northern blotting confirms that hepatic P08833 mRNA abundance also is increased acutely in both P01583 - and P01375 alpha-treated animals . Similar results obtained in adrenalectomized mice indicate that adrenal activation is not required for this effect . Cell culture studies show that cytokines exert direct effects on the production of P08833 by HepG2 hepatoma cells , increasing P08833 levels in conditioned medium and the abundance of P08833 mRNA approximately 3-fold . In contrast , transient transfection studies with P08833 promoter/luciferase reporter gene constructs show that P08833 promoter activity is reduced after 18 hr cytokine treatment . We conclude that P01583 and P01375 alpha increase circulating levels of P08833 , reflecting direct effects on hepatic P08833 mRNA abundance . Stimulation of hepatic P08833 production may contribute to alterations in IGF bioactivity and impaired anabolism in clinical conditions where cytokine production is high . Additional studies are required to identify specific mechanisms mediating effects of cytokines on hepatic production of P08833 . P01308 action on H292 bronchial carcinoma cells as compared to normal bronchial epithelial cells . DB00030 may contribute to bronchial carcinoma due to P08069 activation by high local concentrations . Therefore , effects of insulin and P05019 on human bronchial carcinoma cells ( H292 ) and normal bronchial epithelium cells ( P02100 ) were studied . TGF-β was included since it also influences carcinoma progression . H292 and P02100 cells expressed both the insulin receptor and the P08069 ; in H292 cells an additional , shorter , splicing variant ( IR-A ) of the insulin receptor was present . P06213 expression was around four to five times higher in H292 than in P02100 cells at mRNA and protein levels . P01308 and TGF-β exerted contrary actions on proliferation and gene expression in H292 cells . Genes regulated by insulin , P05019 , and TGF-β were linked to inflammation , cell adhesion , muscle contraction and differentiation . P01308 and P05019 also suppressed DNA repair genes . EC(50) for insulin-induced proliferation was around 5 nM in H292 and around 30 nM P02100 cells . The EC(50) values for gene expression ranged from 9 to 90 nM in both cell types , dependent on the gene studied . In H292 cells , the proliferative response was much stronger if TGF-β was present . In P02100 cells this interaction of insulin and TGF-β was not observed , and changes in gene expression were mostly lower by at least 10-fold as compared to H292 . All in all , the insulin effects in H292 were generally much stronger than in P02100 cells and - with regard to proliferation - occurred at lower concentrations . Thus , insulin will hardly induce cancer from normal bronchial cells but may favour progression of pre-existing tumours . Study of folate receptor genes in nonsyndromic familial and sporadic cleft lip with or without cleft palate cases . DB00158 receptor family members ( FOLRs ) mediate the delivery of 5-methyltetrahydrofolate to the interior of , out of within , or between cells in a process known as potocytosis . Three FOLRs and a pseudogene map to 11q13.4 . The aim of this study was to verify whether FOLRs could be responsible for the onset of nonsyndromic cleft lip with or without cleft palate ( CL/P ) . Linkage and linkage disequilibrium between genetic markers and disorder were analyzed . Patients and their mothers from 71 familial CL/P pedigrees and 75 sporadic cases from Italian population were investigated by PCR-SSCP analysis . Data from mutation scanning allowed us to find only a silent mutation in P15328 present in a mother and her child . Our findings do not support P15328 and P14207 genes in the onset of CL/P . Characterization of alcohol dehydrogenase 1 and 3 from Neurospora crassa FGSC2489 . DB00898 dehydrogenase ( DB00067 ) is a key enzyme in the production and utilization of alcohols . Some also catalyze the formation of carboxylate esters from alcohols and aldehydes . The P07327 and P00326 genes of Neurospora crassa FGSC2489 were cloned and expressed in recombinant Escherichia coli to investigate their alcohol dehydrogenation and carboxylate ester formation abilities . Homology analysis and sequence alignment of amino acid sequence indicated that P07327 and P00326 of N. crassa contained a zinc-binding consensus sequence and a NAD(+)-binding motif and showed 54-75 % identity with fungi ADHs . N. crassa P07327 was expressed in E. coli to give a specific activity of 289 +/- 9 mU/mg using ethanol and NAD(+) as substrate and cofactor , respectively . Corresponding experiments on the expression and activity of P00326 gave 4 mU/mg of specific activity . N. crassa P07327 preferred primary alcohols containing P01024 - Q99618 carbons to secondary alcohols such as 2-propanol and 2-butanol . N. crassa P07327 possessed 5.3 mU/mg of specific carboxylate ester-forming activity accumulating 0.4 mM of ethyl acetate in 18 h . Substrate specificity of various linear alcohols and aldehydes indicated that short chain-length alcohols and aldehydes were good substrates for carboxylate ester production . N. crassa P07327 was a primary alcohol dehydrogenase using cofactor NAD(+) preferably and possessed carboxylate ester-forming activity with short chain alcohols and aldehydes . Lupus-specific kidney deposits of HSP90 are associated with altered IgG idiotypic interactions of anti-HSP90 autoantibodies . Previous studies have shown that autoantibodies to heat shock protein 90 ( HSP90 ) are elevated in a significant proportion of patients with systemic lupus erythematosus ( SLE ) who are more likely to have renal disease and a low P01024 level . Using samples from 24 patients , we searched for glomerular deposits of HSP90 in renal biopsy specimens from seven patients with lupus nephritis and 17 cases of glomerulonephritis from patients without SLE . Positive glomerular immunofluorescent staining for HSP90 was observed in six of seven cases of SLE and positive tubular staining in two of seven SLE patients . The staining for HSP90 was granular in nature and was located in subepithelial , subendothelial and mesangial areas . None of the non-SLE renal biopsies revealed positive staining for HSP90 deposition . Further we showed the presence of anti-HSP90 IgG autoantibodies in IgG from sera of patients with SLE as well as in normal human IgG ( DB00028 ) . In normal IgG this autoreactivity could be adsorbed almost completely on F(ab')2 fragments from the same IgG preparation , coupled to Sepharose and could be inhibited by the effluent obtained after subjecting normal IgG to HSP90 affinity column . These findings indicate that anti-HSP90 natural autoantibodies are blocked by idiotypic interactions within the IgG repertoire . Unlike natural autoantibodies , anti-HSP90 IgG from SLE patients ' sera were only moderately adsorbed on F(ab')2 fragments of normal IgG . These results demonstrate that immunopathogenesis of lupus nephritis is associated with HSP90 ( as an autoantigen ) and that the pathology is associated with altered idiotypic regulation of the anti-HSP90 IgG autoantibodies . 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 . 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 . Analysis of SNP profiles in patients with major depressive disorder . The present study focused on 91 single-nucleotide polymorphisms ( SNPs ) in 21 candidate genes to find associations with major depressive disorder ( MDD ) . In total , 160 healthy controls and 177 patients with MDD were studied . We applied arrayed primer extension ( P27695 ) based genotyping technology followed by association and haplotype analysis . SNPs in P32238 , P21728 , P14416 , and P28335 genes showed nominally significant associations with MDD . None of these associations remained significant after adjustment for multiple testing . Haplotype analysis revealed P32238 haplotypes to be associated with MDD ( global p=0.004 ) . More precisely , we found the GAGT haplotype to be associated with increased risk for MDD ( OR 7.42 , 95 % CI 2.13-25.85 , p=0.002 ) . This haplotype effect remained significant after Bonferroni correction ( p=0.04 after Bonferroni 's adjustment ) . Altogether we were able to find some nominal associations , but due to small sample size these results should be taken as exploratory . However , the effect of GAGT haplotype on the P32238 gene may be considered as increasing the risk for MDD . DB09052 induces autologous T-cell killing of chronic lymphocytic leukemia cells . Chronic lymphocytic leukemia is an incurable B-cell malignancy that is associated with tumor cell-mediated T-cell dysfunction . It therefore represents a challenging disease for T-cell immunotherapeutics . The P15391 /CD3 bi-specific antibody construct blinatumomab ( AMG103 or MT103 ) has been tested clinically in non-Hodgkin 's lymphoma and acute lymphoblastic leukemia but has not been assessed in chronic lymphocytic leukemia . We investigated whether blinatumomab could overcome T-cell dysfunction in chronic lymphocytic leukemia in vitro . DB09052 was tested on peripheral blood mononuclear cells from 28 patients ( treatment naïve and previously treated ) . T-cell activation and function , as well as cytotoxicity against leukemic tumor cells were measured . DB09052 induced T-cell activation , proliferation , cytokine secretion and granzyme B release in a manner similar to that occurring with stimulation with anti-CD3/anti- P10747 beads . However , only blinatumomab was able to induce tumor cell death and this was found to require blinatumomab-mediated conjugate formation between T cells and tumor cells . Cytotoxicity of tumor cells was observed at very low T-cell:tumor cell ratios . A three-dimensional model based on confocal microscopy suggested that up to 11 tumor cells could cluster round each T cell . Importantly , blinatumomab induced cytotoxicity against tumor cells in samples from both treatment-naïve and treated patients , and in the presence of co-culture pro-survival signals . The potent cytotoxic action of blinatumomab on tumor cells appears to involve conjugation of T cells with tumor cells at both the activation and effector stages . The efficacy of blinatumomab in vitro suggests that the bi-specific antibody approach may be a powerful immunotherapeutic strategy in chronic lymphocytic leukemia . Monokine production by human T cells ; P01583 production restricted to memory T cells . The production of cytokines is a key event of inflammation . In this report we demonstrate that normal human T cells are capable to produce P01583 , P05231 , and P01375 , cytokines formerly considered to be monokines . This production was optimal after stimulation with a combination of anti- P06729 , PMA , and anti- P10747 . All three cytokines were produced in a bioactive form . Both naive ( P01730 +CD45RA+ ) and memory ( P01730 +CD45RO+ ) subsets of T cells were shown to produce similar amounts of P05231 and P01375 . In contrast the production of P01583 was found to be completely restricted to the P01730 +CD45RO+ subset . The finding that T cells are such potent producers of these important mediators of the inflammatory response might be a key observation in the appreciation of the role of T cells in chronic inflammation . Selective P36888 inhibition of P36888 -ITD+ acute myeloid leukaemia resulting in secondary D835Y mutation : a model for emerging clinical resistance patterns . Acquired resistance to selective P36888 inhibitors is an emerging clinical problem in the treatment of P36888 -ITD(+) acute myeloid leukaemia ( AML ) . The paucity of valid pre-clinical models has restricted investigations to determine the mechanism of acquired therapeutic resistance , thereby limiting the development of effective treatments . We generated selective P36888 inhibitor-resistant cells by treating the P36888 -ITD(+) human AML cell line MOLM-13 in vitro with the P36888 -selective inhibitor MLN518 , and validated the resistant phenotype in vivo and in vitro . The resistant cells , MOLM-13-RES , harboured a new D835Y tyrosine kinase domain ( TKD ) mutation on the P36888 -ITD(+) allele . Acquired TKD mutations , including D835Y , have recently been identified in P36888 -ITD(+) patients relapsing after treatment with the novel P36888 inhibitor , DB05213 . Consistent with this clinical pattern of resistance , MOLM-13-RES cells displayed high relative resistance to DB05213 and DB00398 . Furthermore , treatment of MOLM-13-RES cells with DB05213 lead to loss of the P36888 wild-type allele and the duplication of the P36888 -ITD-D835Y allele . Our P36888 -Aurora kinase inhibitor , CCT137690 , successfully inhibited growth of P36888 -ITD-D835Y cells in vitro and in vivo , suggesting that dual P36888 -Aurora inhibition may overcome selective P36888 inhibitor resistance , in part due to inhibition of Aurora kinase , and may benefit patients with P36888 -mutated AML . Vitamin D metabolism , mechanism of action , and clinical applications . DB00169 is made in the skin from 7-dehydrocholesterol under the influence of UV light . DB00153 ( ergocalciferol ) is derived from the plant sterol ergosterol . Vitamin D is metabolized first to 25 hydroxyvitamin D ( 25OHD ) , then to the hormonal form 1,25-dihydroxyvitamin D ( 1,25(OH)2D ) . Q6VVX0 is the most important 25-hydroxylase ; O15528 is the key 1-hydroxylase . Both 25OHD and 1,25(OH)2D are catabolized by Q07973 . 1,25(OH)2D is the ligand for the vitamin D receptor ( P11473 ) , a transcription factor , binding to sites in the DNA called vitamin D response elements ( VDREs ) . There are thousands of these binding sites regulating hundreds of genes in a cell-specific fashion . P11473 -regulated transcription is dependent on comodulators , the profile of which is also cell specific . Analogs of 1,25(OH)2D are being developed to target specific diseases with minimal side effects . This review will examine these different aspects of vitamin D metabolism , mechanism of action , and clinical application . Cholecystokinin ( CCK ) stimulates aldosterone secretion from human adrenocortical cells via CCK2 receptors coupled to the adenylate cyclase/protein kinase A signaling cascade . Cholecystokinin ( CCK ) IS a regulatory peptide that acts via two receptor subtypes , P32238 and P32239 . RT-PCR demonstrated the expression of both P32238 and P32239 in the zona glomerulosa ( ZG ) , but not zona fasciculata-reticularis cells of the human adrenal cortex . CCK and the P32239 agonist pentagastrin enhanced basal aldosterone secretion from ZG cells without affecting cortisol production from zona fasciculata-reticularis cells . The aldosterone response to CCK and pentagastrin was suppressed by a P32239 antagonist , but not by a P32238 antagonist . DB00183 evoked a sizeable DB02527 , but not inositol triphosphate , response from ZG cells , whereas CCK plus P32239 antagonist was ineffective . The DB02527 response to pentagastrin was abrogated by P32239 antagonist or the adenylate cyclase inhibitor SQ-22536 , and the aldosterone response was abolished by both SQ-22536 and the protein kinase A inhibitor H-89 . Both CCK and pentagastrin increased steroidogenic acute regulatory protein mRNA expression in ZG cells ; the effect was abrogated by P32239 antagonist . We conclude that CCK exerts secretagogue action on human ZG cells , acting through CCK2-Rs coupled to the adenylate cyclase/protein kinase A signaling cascade , which , in turn , stimulates the expression of steroidogenic acute regulatory protein , the rate-limiting step of steroidogenesis . 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 . Single and multigenic analysis of the association between variants in 12 steroid hormone metabolism genes and risk of prostate cancer . To estimate the prostate cancer risk conferred by individual single nucleotide polymorphisms ( SNPs ) , SNP-SNP interactions , and/or cumulative SNP effects , we evaluated the association between prostate cancer risk and the genetic variants of 12 key genes within the steroid hormone pathway ( P05093 , P37058 , P03372 , P31213 , P14060 , P26439 , P11511 , P04798 , Q16678 , P08684 , O15528 , and Q07973 ) . A total of 116 tagged SNPs covering the group of genes were analyzed in 2,452 samples ( 886 cases and 1,566 controls ) in three ethnic/racial groups . Several SNPs within P11511 were significantly associated with prostate cancer in all three ethnicities ( P = 0.001-0.009 ) . Genetic variants within P26439 and Q07973 conferred increased risk of prostate cancer in non-Hispanic or Hispanic Caucasians . A significant gene-dosage effect for increasing numbers of potential high-risk genotypes was found in non-Hispanic and Hispanic Caucasians . Higher-order interactions showed a seven-SNP interaction involving P37058 , P11511 , and Q07973 in Hispanic Caucasians ( P = 0.001 ) . In African Americans , a 10-locus model , with SNPs located within P31213 , P37058 , P05093 , O15528 , P11511 , and Q07973 , showed a significant interaction ( P = 0.014 ) . In non-Hispanic Caucasians , an interaction of four SNPs in P26439 , P37058 , and P11511 was found ( P < 0.001 ) . These data are consistent with a polygenic model of prostate cancer , indicating that multiple interacting genes of the steroid hormone pathway confer increased risk of prostate cancer . Hematopoietic support and cytokine expression of murine-stable hepatocyte cell lines ( O15527 ) . It was recently reported that transgenic expression in the liver of truncated human DB00134 renders hepatocytes constitutively resistant to apoptosis and reproducibly permits their immortalization . The derived stable cell lines ( O15527 from DB00134 murine hepatocyte ) are highly differentiated and nontransformed . In this report , the capacity of MMHs to support in vitro hematopoiesis is characterized . By reverse-transcription polymerase chain reaction , the expression by MMHs of cytokines involved in the survival and self-renewal of early progenitor cells ( stem cell factor and P36888 ligand ) as well as those acting at different stages of progenitor differentiation ( interleukin [ IL ] 1beta , P08700 , leukemia inhibitory factor , P05231 , granulocyte-macrophage colony-stimulating factor , granulocyte colony-stimulating factor , macrophage colony-stimulating factor , and thrombopoietin ) was shown . A ribonuclease protection assay further substantiated the presence of at least six cytokine transcripts in O15527 lines . Cocultures between O15527 layers and progenitor-enriched fetal liver hematopoietic cells resulted in a 40-fold to 80-fold expansion of total hematopoietic cells and in a 2.5-fold expansion of clonogenic progenitors after 1 to 2 weeks . Hematopoiesis was maintained for up to 6 weeks with formation of typical cobblestone cell areas and continuous differentiation of precursor into cells at various degrees of maturation . At 5 weeks of coculture , clonogenic progenitors were maintained at 20 % of the input level in coculture with embryonic-derived hepatocytes , showing the ability of hepatocyte feeder layer to support survival and possibly self-renewal of clonogenic progenitors . Therefore , the data emphasize a direct role of the hepatocyte in sustaining hematopoietic cell proliferation and differentiation . Inhibitory effect of the P35462 on insulin receptor expression and function in vascular smooth muscle cells . BACKGROUND : Vascular smooth muscle cell ( VSMC ) proliferation is regulated by numerous hormones and humoral factors . Our previous study found that stimulation of D(1)-like dopamine receptors inhibited insulin receptor expression and function in VSMCs . We hypothesize that there is also an interaction between D(3) dopamine and insulin receptors , i.e. , stimulation of the D(3) receptor inhibits insulin receptor expression and function . METHODS : Receptor expression was determined by immunoblotting , immunohistochemisty , and reverse transcriptase-PCR ; VSMC proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-diphenyl-tetrazolium bromide ( MTT ) assay and cell number . RESULTS : P06213 protein is increased in the aorta of D(3) receptor deficient mice . Stimulation of the D(3) receptor inhibited insulin receptor mRNA and protein expression and insulin-mediated VSMC proliferation , and increased protein kinase A ( PKA ) activity , insulin receptor phosphorylation , and degradation in immortalized aortic VSMCs ( A10 cells ) . These effects were blocked by a PKA inhibitor , indicating that the D(3) receptor-mediated decrease in insulin receptor expression was related to a decrease in transcription/post-transcription and increased degradation , involving PKA signaling . CONCLUSIONS : D(3) receptor stimulation may be a target to reduce the adverse effect of insulin in hypertension by inhibition of insulin receptor expression and function in arterial VSMCs .

MH_train_1314

MH_train_1314

MH_train_1314

interacts_with DB08881?

multiple_choice

Opposing actions of extracellular signal-regulated kinase ( P29323 ) and signal transducer and activator of transcription 3 ( P40763 ) in regulating microtubule stabilization during cardiac hypertrophy . Excessive proliferation and stabilization of the microtubule ( MT ) array in cardiac myocytes can accompany pathological cardiac hypertrophy , but the molecular control of these changes remains poorly characterized . In this study , we examined MT stabilization in two independent murine models of heart failure and revealed increases in the levels of post-translationally modified stable MTs , which were closely associated with P40763 activation . To explore the molecular signaling events contributing to control of the cardiac MT network , we stimulated cardiac myocytes with an α-adrenergic agonist phenylephrine ( PE ) , and observed increased tubulin content without changes in detyrosinated ( glu-tubulin ) stable MTs . In contrast , the hypertrophic interleukin-6 ( P05231 ) family cytokines increased both the glu-tubulin content and glu-MT density . When we examined a role for P29323 in regulating cardiac MTs , we showed that the MEK/ P29323 -inhibitor U0126 increased glu-MT density in either control cardiac myocytes or following exposure to hypertrophic agents . Conversely , expression of an activated Q02750 mutant reduced glu-tubulin levels . Thus , P29323 signaling antagonizes stabilization of the cardiac MT array . In contrast , inhibiting either O60674 with AG490 , or P40763 signaling with Stattic or siRNA knockdown , blocked cytokine-stimulated increases in glu-MT density . Furthermore , the expression of a constitutively active P40763 mutant triggered increased glu-MT density in the absence of hypertrophic stimulation . Thus , P40763 activation contributes substantially to cytokine-stimulated glu-MT changes . Taken together , our results highlight the opposing actions of P40763 and P29323 pathways in the regulation of MT changes associated with cardiac myocyte hypertrophy . The kinase inhibitor staurosporine induces P55008 arrest at two points : effect on retinoblastoma protein phosphorylation and cyclin-dependent kinase 2 in normal and transformed cells . DB02010 ( ST ) , a protein kinase inhibitor , at a concentration of 20 nM arrests normal diploid fibroblasts 3 h into P55008 ( H. A. Crissman et al. , Proc. Natl. Acad. Sci. USA , 88 : 7580-7584 , 1991 ; K. Abe et al. , Exp. Cell Res. , 192 : 122-127 , 1991 ) . ST ( 2 nM ) induces a new P55008 arrest point at 6 h into P55008 . Partial phosphorylation of the retinoblastoma protein was observed at the 2 nM ST arrest point , whereas the retinoblastoma protein was unphosphorylated or underphosphorylated at the 20 nM arrest point . This correlated with the activity of the cyclin-dependent kinase 2 ( P24941 ) and the phosphorylation of the Thr160 residue of p33CDK2 . The cyclin E and cyclin D1/2 levels were reduced at the 20 nM ST arrest point . In HeLa cells that do not arrest in P55008 in response to 2 or 20 nM ST , the retinoblastoma protein and P24941 phosphorylations and P24941 activity were not affected by ST . These results suggest that ST inhibits one or more P55008 -regulating protein kinases , which lie upstream of P24941 . Glutamatergic regulation of the p70S6 kinase in primary mouse neurons . Brief glutamatergic stimulation of neurons from fetal mice , cultured in vitro for 6 days , activates the P42345 -S6 kinase , P27361 /2 and Akt pathways , to an extent approaching that elicited by brain-derived neurotrophic factor . In contrast , sustained glutamatergic stimulation inhibits P29323 , Akt , and S6K . Glutamatergic activation of S6K is calcium/calmodulin-dependent and is prevented by inhibitors of calcium/calmodulin-dependent protein kinase 2 , phosphatidylinositol 3-OH-kinase and by rapamycin . 2-Amino-5-phosphonovaleric acid , an inhibitor of N'-methyl-D-aspartate receptors , abolishes glutamatergic activation of P27361 /2 but not the activation of P42345 -S6K ; the latter is completely abolished by inhibitors of voltage-dependent calcium channels . Added singly , dopamine gives slight , and norepinephrine a more significant , activation of P29323 and S6K ; both catecholeamines , however , enhance glutamatergic activation of S6K but not P29323 . After 12 days in culture , the response to direct glutamatergic activation is attenuated but can be uncovered by suppression of gamma-aminobutyric acid interneurons with bicuculline in the presence of the weak K(+) channel blocker 4-aminopyridine ( DB06637 ) . This selective synaptic activation of P42345 -S6K is also resistant to APV and inhibited by Ca(2+) channel blockers and higher concentrations of glutamate . P13639 ( P13639 ) is phosphorylated and inhibited by the eEF2 kinase ( P62158 kinase III ) ; the latter is inhibited by the S6K or Rsk . DB11562 / DB06637 or DB00761 -induced depolarization reduces , whereas higher concentrations of glutamate increases , P13639 phosphorylation . Thus the P42345 -S6K pathway in neurons , a critical component of the late phase of LTP , is activated by glutamatergic stimulation in a calcium/calmodulin-dependent fashion through a calcium pool controlled by postsynaptic voltage-dependent calcium channels , whereas sustained stimulation of extrasynaptic glutamate receptors is inhibitory . MEK inhibition potentiates the activity of Hsp90 inhibitor 17- P29372 against pancreatic cancer cells . The Ras/Raf/MEK/ P29323 signaling has been implicated in uncontrolled cell proliferation and tumor progression in pancreatic cancer . The purpose of this study is to evaluate the antitumor activity of MEK inhibitor U0126 in combination with Hsp90 inhibitor 17-allylamino-17-demethoxygeldanamycin ( 17- P29372 ) in pancreatic cancer cells . Western blotting showed that 17- P29372 caused a 2- to 3-fold transient activation of MEK/ P29323 signaling in pancreatic cancer cells . The activation sustained for 6 h before phospho- P29323 ( p- P29323 ) destabilization . The selective MEK inhibitor U0126 completely abolished 17- P29372 induced P27361 /2 activation and resulted in more than 80 % of phospho- P29323 degradation after only 15 min treatment . Moreover , U0126 had complementary effect on 17- P29372 regulated oncogenic and cell cycle related proteins . Although 17- P29372 downregulated cyclin D1 , cyclin E , P11802 and Q00534 , it led to cyclin A and P24941 accumulation , which was reversed by the addition of U0126 . Antiproliferation assay showed that combination of U0126 and 17- P29372 resulted in synergistic cytotoxic effect . More importantly , 17- P29372 alone only exhibited moderate inhibition of cell migration in vitro , while addition of U0126 dramatically enhanced the inhibitory effect by 2- to 5-fold . Taken together , these data demonstrate that MEK inhibitor U0126 potentiates the activity of Hsp90 inhibitor 17- P29372 against pancreatic cancer cells . The combination of Hsp90 and MEK inhibition could provide a promising avenue for the treatment of pancreatic cancer . The small GTPase Rap1 is required for Mn(2+)- and antibody-induced LFA-1- and VLA-4-mediated cell adhesion . In T-lymphocytes the Ras-like small GTPase Rap1 plays an essential role in stimulus-induced inside-out activation of integrin LFA-1 ( alpha(L)beta(2) ) and VLA-4 ( alpha(4)beta(1) ) . Here we show that Rap1 is also involved in the direct activation of these integrins by divalent cations or activating antibodies . Inhibition of Rap1 either by Rap P20936 ( RapGAP ) or the Rap1 binding domain of Q12967 abolished both Mn(2+)- and KIM185 ( anti-LFA-1 ) -induced LFA-1-mediated cell adhesion to intercellular adhesion molecule 1 . Mn(2+)- and TS2/16 ( anti-VLA-4 ) -induced VLA-4-mediated adhesion were inhibited as well . Interestingly , both Mn(2+) , KIM185 and TS2/16 failed to induce elevated levels of Rap1GTP . These findings indicate that available levels of GTP-bound Rap1 are required for the direct activation of LFA-1 and VLA-4 . Pharmacological inhibition studies demonstrated that both Mn(2+)- and KIM185-induced adhesion as well as Rap1-induced adhesion require intracellular calcium but not signaling activity of the MEK- P29323 pathway . Moreover , functional calmodulin signaling was shown to be a prerequisite for Rap1-induced adhesion . From these results we conclude that in addition to stimulus-induced inside-out activation of integrins , active Rap1 is required for cell adhesion induced by direct activation of integrins LFA-1 and VLA-4 . We suggest that Rap1 determines the functional availability of integrins for productive binding to integrin ligands . S-Adenosylmethionine regulates connexins sub-types expressed by hepatocytes . Intercellular communication via P20936 Junctions plays an important role in tissue homeostasis , apoptosis , carcinogenesis , cell proliferation and differentiation . Hepatocyte connexins ( Cx ) 26 and 32 levels are decreased during the de-differentiation process of primary hepatocytes in culture , a situation that is also characterized by a decrease in S-Adenosylmethionine ( DB00118 ) levels . In this current study , we show that DB00118 supplementation in cultured hepatocytes every 12h , leads to an up-regulation of P29033 and 32 mRNA and protein levels and blocks culture-induced P17302 expression , although it failed to increase P29033 and 32 membrane localization and P20936 junction intracellular communication . DB00118 reduced nuclear β-catenin accumulation , which is known to stimulate the TCF/LEF-dependent gene transcription of P17302 . Moreover DB00118 -induced reduction in P17302 and β-catenin was prevented by the proteasome inhibitor MG132 , and was not mediated by GSK3 activity . DB00118 , and its metabolite 5'-methylthioadenosine ( MTA ) increased P29033 mRNA in a process partially mediated by DB00640 A(2A) receptors but independent of PKA . Finally livers from Q00266 knockout mice , characterized by low hepatic DB00118 levels , express higher P17302 and lower P29033 and 32 protein levels than control mice . These results suggest that DB00118 maintains a characteristic expression pattern of the different Cxs in hepatocytes by differentially regulating their levels . Adaptive responses to dasatinib-treated lung squamous cell cancer cells harboring Q16832 mutations . Q16832 mutations occur in approximately 4 % of lung squamous cell cancer ( SCC ) where the tyrosine kinase inhibitor dasatinib has emerged as a new therapeutic option . We found that P29323 and AKT phosphorylation was weakly inhibited by dasatinib in Q16832 -mutant lung SCC cells , suggesting that dasatinib inhibits survival signals distinct from other oncogenic receptor tyrosine kinases ( RTK ) and/or compensatory signals exist that dampen dasatinib activity . To gain better insight into dasatinib 's action in these cells , we assessed altered global tyrosine phosphorylation ( pY ) after dasatinib exposure using a mass spectrometry-based quantitative phosphoproteomics approach . Overlaying protein-protein interaction relationships upon this dasatinib-regulated pY network revealed decreased phosphorylation of Src family kinases and their targets . Conversely , dasatinib enhanced tyrosine phosphorylation in a panel of RTK and their signaling adaptor complexes , including P00533 , MET/ Q13480 , and P08069 / Q9Y4H2 , implicating a RTK-driven adaptive response associated with dasatinib . To address the significance of this observation , these results were further integrated with results from a small-molecule chemical library screen . We found that dasatinib combined with MET and insulin-like growth factor receptor ( P08069 ) inhibitors had a synergistic effect , and ligand stimulation of P00533 and MET rescued Q16832 -mutant lung SCC cells from dasatinib-induced loss of cell viability . Importantly , we observed high levels of tyrosine-phosphorylated P00533 and MET in a panel of human lung SCC tissues harboring Q16832 mutations . Our results highlight potential RTK-driven adaptive-resistant mechanisms upon Q16832 targeting , and they suggest new , rationale cotargeting strategies for Q16832 -mutant lung SCC . Enhancement of L-cystine transport activity and its relation to Q9UPY5 gene induction at the blood-brain barrier by diethyl maleate treatment . The purpose of the present study was to elucidate the mechanism of enhancement of L-cystine uptake at the blood-brain barrier ( BBB ) . The uptake of [(14)C]L-cystine and [(3)H]L-glutamic acid ( L- DB00142 ) was determined using a mouse brain endothelial cell line ( MBEC4 ) as an in vitro BBB model . The mRNA levels of L-cystine/L- DB00142 exchanger , system x(c)(-) , which consists of Q9UPY5 and P08195 , were determined by quantitative real-time reverse transcription-polymerase chain reaction analysis . The [(14)C]L-cystine uptake by MBEC4 cells appeared to be mediated via an Na(+)-independent saturable process . The corresponding Michaelis-Menten constant ( K(m) ) was 63.7 microM . In the presence of L- DB00142 , there was competitive inhibition with an inhibition constant ( K(i) ) of 83.5 microM . [(3)H]L- DB00142 uptake in the absence of Na(+) was saturable with a K(m) of 48.1 microM , and it exhibited competitive inhibition with a K(i) of 24.9 microM in the presence of L-cystine . The mutual inhibition between L-cystine and L- DB00142 and the type of inhibition suggest that system x(c)(-) operates in MBEC4 cells . The Q9UPY5 and P08195 mRNAs were expressed in MBEC4 cells and , following diethyl maleate ( DEM ) treatment , the Q9UPY5 mRNA level and L-cystine uptake in MBEC4 cells were enhanced in parallel with an increase in DEM concentration ( up to 500 microM ) . Concomitantly , the glutathione concentration in MBEC4 cells was increased . In conclusion , system x(c)(-)-mediated L-cystine uptake takes place in MBEC4 cells . DB00138 transport via system x(c)(-) at the BBB is likely to be induced under oxidative stress conditions following DEM treatment due to enhanced transcription of the Q9UPY5 gene . DB11320 Promotes the Release of P05231 via the P35367 /p38 and NF-κB Pathways in Nasal Fibroblasts . PURPOSE : Based on the close relationship between histamine and interleukin 6 ( P05231 ) , we hypothesized that histamine may regulate the production of cytokines , such as P05231 , during allergic inflammation . Here , we examined the role of histamine in P05231 production and histamine receptor activity in nasal fibroblasts , along with the mechanisms underlying these effects . METHODS : Experiments were performed using nasal fibroblasts from 8 normal patients . RT-PCR was used to identify the major histamine receptors expressed in nasal fibroblasts . Fibroblasts were then treated with histamine with or without histamine-receptor antagonists , and monitored for P05231 production using an ELISA . Four potential downstream signaling molecules , p38 , extracellular signal-regulated kinase ( P29323 ) , c-Jun N-terminal kinase ( JNK ) , and NF-κB , were evaluated by Western blot , and a luciferase reporter assay . RESULTS : Elevated expression was seen for all histamine receptors , with P05231 protein levels increasing significantly following histamine stimulation . Among the histamine-receptor specific antagonists , only the P35367 antagonist significantly decreased P05231 production in histamine-stimulated nasal fibroblasts . DB11320 increased the expression level of phosphorylated p38 ( pp38 ) , pERK , and pJNK , as well as NF-κB induction . The P35367 antagonist actively suppressed pp38 and NF-κB expression in histamine-induced nasal fibroblasts , but not pERK and pJNK . The p38 inhibitor strongly attenuated P05231 production in histamine-stimulated nasal fibroblasts . CONCLUSIONS : The data presented here suggest that antihistamines may be involved in the regulation of cytokines , such as P05231 , due to the role of histamine as an inflammatory mediator in nasal fibroblasts . P01308 -like growth factor-I is a serum component stimulating growth of human neuroblastoma . Human non-autocrine neuroblastoma cells SK-N-SH and LF require serum for proliferation in vitro . We wished to determine the role of serum-borne insulin-like growth factor I ( P05019 ) as mitogen for these cells . Introduction of the monoclonal antibody alpha-IR3 against human P08069 reduced proliferation in the presence of fetal bovine serum ( FBS ) . P05019 ( 5 nM ) was as effective as FBS ( 10 % ) in stimulating proliferation . DB00071 mimicked the effects of P05019 , but at a 1000-fold higher concentration . The antibody alpha-IR3 reduced growth stimulated by P05019 more effectively than growth stimulated by insulin . Thus , proliferation of human non-autocrine neuroblastoma cells can be effectively manipulated by exogenous P05019 . A new epigenetic challenge : systemic lupus erythematosus . In recent years , compelling evidence has been gathered that supports a role for epigenetic alterations in the pathogenesis of systemic lupus erythematosus ( SLE ) . Different blood cell populations of SLE patients are characterized by a global loss of DNA methylation . This process is associated with defects in P29323 pathway signalling and consequent P26358 1 downregulation . Hypomethylation of gene promoters has been described , which permits transcriptional activation and therefore functional changes in the cells and also hypomethylation of the ribosomal RNA gene cluster . Among the identified targets undergoing demethylation are genes involved in autoreactivity ( P20701 ) , osmotic lysis and apoptosis ( P14222 , P50281 and P80188 ) , antigen presentation ( Q99062 ) , inflammation ( MMP 14 ) , B- T-cell interaction ( P32970 and P29965 ) and cytokine pathways ( Q99062 , P05112 , P05231 and P38484 ) . DNA methylation inhibitors are also known to induce autoreactivity in vitro and cause a lupus-like disease in vivo . Further , altered patterns of histone modifications have been described in SLE . P01730 + lymphocytes undergo global histone H3 and H4 deacetylation and consequent skewed gene expression . Although multiple lines of evidence highlight the contribution of epigenetic alterations to the pathogenesis of lupus in genetically predisposed individuals , many questions remain to be answered . Attaining a deeper understanding of these matters will create opportunities in the promising area of epigenetic treatments . Acute ethanol preexposure promotes liver regeneration after partial hepatectomy in mice by activating P05091 . It is known that chronic ethanol significantly impairs liver regeneration . However , the effect of acute ethanol exposure on liver regeneration remains largely unknown . To address this question , C57Bl6/J mice were exposed to acute ethanol ( 6 g/kg intragastrically ) for 3 days , and partial hepatectomy ( PHx ) was performed 24 h after the last dose . Surprisingly , acute ethanol preexposure promoted liver regeneration . This effect of ethanol did not correlate with changes in expression of cell cycle regulatory genes ( e.g. , cyclin D1 , P38936 , and p27 ) but did correlate with protection against the effect of PHx on indices of impaired lipid and carbohydrate metabolism . DB00898 preexposure protected against inhibition of the oxidant-sensitive mitochondrial enzyme , aconitase . The activity of aldehyde dehydrogenase 2 ( P05091 ) was significantly increased by ethanol preexposure . The effect of ethanol was blocked by inhibiting ( DB02115 ) and was mimicked by activating ( Alda-1 ) P05091 . Lipid peroxides are also substrates for P05091 ; indeed , alcohol preexposure blunted the increase in lipid peroxidation ( 4OH-nonenal adducts ) caused by PHx . Taken together , these data suggest that acute preoperative ethanol exposure " preconditions " the liver to respond more rapidly to regenerate after PHx by activating mitochondrial P05091 , which prevents oxidative stress in this compartment . P15056 inhibitors suppress apoptosis through off-target inhibition of JNK signaling . DB08881 and dabrafenib selectively inhibit the P15056 ( P15056 ) kinase , resulting in high response rates and increased survival in melanoma . Approximately 22 % of individuals treated with vemurafenib develop cutaneous squamous cell carcinoma ( cSCC ) during therapy . The prevailing explanation for this is drug-induced paradoxical P29323 activation , resulting in hyperproliferation . Here we show an unexpected and novel effect of vemurafenib/PLX4720 in suppressing apoptosis through the inhibition of multiple off-target kinases upstream of c-Jun N-terminal kinase ( JNK ) , principally Q9NYL2 . JNK signaling is suppressed in multiple contexts , including in cSCC of vemurafenib-treated patients , as well as in mice . Expression of a mutant Q9NYL2 that can not be inhibited reverses the suppression of JNK activation and apoptosis . Our results implicate suppression of JNK-dependent apoptosis as a significant , independent mechanism that cooperates with paradoxical P29323 activation to induce cSCC , suggesting broad implications for understanding toxicities associated with P15056 inhibitors and for their use in combination therapies . DOI : http://dx.doi.org/10.7554/eLife.00969.001 . Antihistamine effects on prefrontal cortex activity during working memory process in preschool children : a near-infrared spectroscopy ( NIRS ) study . P35367 antagonists ( antihistamines ) are widely used for the treatment of allergic disorders in young children . This study examined the effects of antihistamine on prefrontal cortex activity in preschool children using near-infrared spectroscopy ( NIRS ) , an emerging brain-imaging method suitable for psychological experiments , especially in young children . We examined the changes of oxygenated hemoglobin concentration in the prefrontal cortex while children performed a spatial working memory task , 3h after taking a first-generation antihistamine ( ketotifen ) , second-generation antihistamine ( epinastine ) , or placebo . Fifteen healthy preschool children ( mean age , 5.5 years ) participated . DB00920 significantly impaired behavioral performance and cortical activation at the lateral prefrontal cortex in the working memory task , compared with epinastine and placebo . There were no sedative effects on neural response or behavioral performance after epinastine administration . This paper demonstrates for the first time differential sedation effects of first- and second-generation antihistamines on brain hemodynamic response in young children . Also discussed is the utility of the NIRS technique in neuropsychopharmacological studies of children . 5-Aza-2'-deoxycitydine induces demethylation and up-regulates transcription of P42771 gene in human gastric cancer cell lines . BACKGROUND : To investigate the effects of DNA methylation on the expression of tumor-associated genes and the cell cycle in human gastric cancer cells . METHODS : Two gastric cancer cell lines ( MKN-45 and HGC-27 ) were treated with DNA methyltransferase ( P26358 ) inhibitor , DB01262 ( 5-aza-dC ) . The expressions of P42771 , p21WAF1 , p53 , p73 , c-Ha-ras and c-myc genes mRNA were detected by using reverse transcription PCR ( RT-PCR ) . DNA methylation status of P42771 gene promoter was assayed by bisulfite modification and sequencing . The cell cycle was analyzed by using flow cytometry ( DB00828 ) . RESULTS : 5-aza-dC induced the demethylation of P42771 gene promoter . The expression of P42771 mRNA was obviously up-regulated by treatment with 10 micro mol/L ( MKN-45 cells ) or 5 micro mol/L ( HGC-27 cells ) of 5-aza-dC for 24 hours . However , 5-aza-dC treatment failed to regulate the expressions of p21WAF1 , p53 , p73 , c-Ha-ras and c-myc genes in MKN-45 and HGC-27 cells . Furthermore , 5-aza-dC induced the cell cycle arrest in P55008 phase in HGC-27 cell , but not in MKN-45 cell . CONCLUSIONS : DNA methylation regulates the transcription of P42771 but not p21WAF1 and proto-oncogenes in human gastric cancer cell lines MKN-45 and HGC-27 . Overexpression of nuclear distribution protein ( hNUDC ) causes pro-apoptosis and differentiation in Dami megakaryocytes . OBJECTIVES : Overexpression of hNUDC , a member of the nuclear distribution protein family , reduces cell population growth in prostate cancer cell lines , concurrent with induced morphological change and enhanced polyploidization . These phenomena are also closely associated with terminal phases of megakaryocyte maturation . MATERIALS AND METHODS : In Dami cells , MTT and trypan blue assays were used to investigate cell viability and proliferation effects of hNUDC , and flow cytometry was used to analyse cell cycle and DNA content . Real-time RT-PCR was employed to detect mRNA expression . Activations of caspase-3 , P29323 , Akt and Stat-5 were determined by immunoblotting . May-Grünwald-Giemsa staining was performed to reveal cell morphology . RESULTS AND CONCLUSION : Functional studies using adenovirus-mediated hNUDC overexpression led to inhibition of megakaryocyte proliferation via cell cycle arrest in G2/M transition phase . This process could have been be mediated by upregulation of P38936 and downregulation of its downstream targets , including cyclin B1 , cyclin B2 and c-myc . Enhanced apoptosis in turn ensued , characterized by increased caspase-3 activation , upregulation of pro-apoptotic Bax and downregulation of anti-apoptotic Bcl-2 . Furthermore , hNUDC overexpression elevated the level of megakaryocyte maturation , associated with increased polyploidy , cell morphological changes and increased expression of cell surface differentiation markers , including CD10 , P16070 , CD41 and CD61 . Our results further suggest that the P29323 signalling pathway was involved in hNUDC overexpression-induced apoptosis . Taken together , this study provides experimental evidence for overexpression of hNUDC in Dami cells and suggests that activation of apoptotic machinery may be involved in megakaryocytic differentiation . A phase 2 , randomized , double-blind , placebo-controlled study of siltuximab ( anti- P05231 mAb ) and bortezomib versus bortezomib alone in patients with relapsed or refractory multiple myeloma . We compared the safety and efficacy of siltuximab ( S ) , an anti-interleukin-6 chimeric monoclonal antibody , plus bortezomib ( B ) with placebo ( plc ) + B in patients with relapsed/refractory multiple myeloma in a randomized phase 2 study . DB09036 was given by 6 mg/kg IV every 2 weeks . On progression , B was discontinued and high-dose dexamethasone could be added to S/plc . Response and progression-free survival ( PFS ) were analyzed pre-dexamethasone by European Group for Blood and Marrow Transplantation ( EBMT ) criteria . For the 281 randomized patients , median PFS for S + B and plc + B was 8.0 and 7.6 months ( HR 0.869 , P = 0.345 ) , overall response rate was 55 versus 47 % ( P = 0.213 ) , complete response rate was 11 versus 7 % , and median overall survival ( OS ) was 30.8 versus 36.8 months ( HR 1.353 , P = 0.103 ) . Sustained suppression of P02741 , a marker reflective of inhibition of interleukin-6 activity , was seen with S + B . DB09036 did not affect B pharmacokinetics . DB09036 /placebo discontinuation ( 75 versus 66 % ) , grade ≥3 neutropenia ( 49 versus 29 % ) , thrombocytopenia ( 48 versus 34 % ) , and all-grade infections ( 62 versus 49 % ) occurred more frequently with S + B . The addition of siltuximab to bortezomib did not appear to improve PFS or OS despite a numerical increase in response rate in patients with relapsed or refractory multiple myeloma . DB00149 stimulates Q15758 amino acid transporter expression in porcine jejunal epithelial cell line ( IPEC-J2 ) through PI3K/Akt/ P42345 and P29323 signaling pathways . DB00149 has been shown to influence intestinal protein metabolism , cell proliferation and migration . Furthermore , our previous study demonstrated that branched-chain amino acids could modulate the intestinal amino acid and peptide transporters in vivo . As the possible mechanisms are still largely unknown , in the present work , we studied the transcriptional and translational regulation of leucine on amino acid transporter production in IPEC-J2 cells and the signaling pathways involved . Treatment of IPEC-J2 cells with 7.5 mM leucine enhanced the mRNA expression of the Na(+)-neutral AA exchanger 2 ( Q15758 ) and 4F2 heavy chain ( P08195 ) and caused an increase in Q15758 protein expression . DB00149 also activated phosphorylation of Q13541 and P06730 through the phosphorylation of P42345 , Akt and P29323 signaling pathways in IPEC-J2 cells . Pre-treatment of IPEC-J2 cells with inhibitors of P42345 and Akt ( rapamycin and wortmannin ) or an inhibitor of P29323 ( PD098059 ) for 30 min before leucine treatment attenuated the positive effect of leucine in enhancing the protein abundance of Q15758 . These results demonstrate that leucine could up-regulate the expression of the amino acid transporters ( Q15758 ) through transcriptional and translational regulation by P29323 and PI3K/Akt/ P42345 activation . Site-specific mutagenesis of the histidine precursor of diphthamide in the human elongation factor-2 gene confers resistance to diphtheria toxin . Protein synthesis elongation factor 2 ( P13639 ) from eukaryotes contains a conserved post-translationally modified histidine residue known as diphthamide . DB03223 is a unique site of ADP-ribosylation by diphtheria toxin ( DT ) , which is responsible for cell killing . In this report , we describe the construction of DT-resistant HeLa cell lines by engineering the toxin-resistant form of its specific substrate , protein elongation factor-2 . Using site-specific mutagenesis of the histidine precursor of diphthamide , the histidine residue of codon 715 in human P13639 cDNA was substituted with one of four amino acid residue codons : leucine , methionine , asparagine or glutamine . Mutant EF-2s were subcloned into a pCMVexSVneo expression vector , transfected into HeLa cells , and DT-resistant cell clones were isolated . The protective effect of mutant EF-2s against cell killing by DT , after exposing all four mutant strains derived from HeLa cells to different concentrations of the toxin ( 5-20 ng/mL ) was demonstrated by : ( 1 ) the normal morphological appearance of the cells ; ( 2 ) their unaffected or slightly slower growth rates ; ( 3 ) their undisturbed electrophoretic DNA profiles whose integrity was virtually preserved . Mutant cell strains showed also considerable levels of resistance to very high concentrations of DT , in that they maintained slower but consistent rates of cell growth . It was hence concluded that despite its strict conservation and unique modification , the diphthamide histidine appears not to be essential to the function of human P13639 in protein synthesis . In addition , DT-resistant HeLa cell clones should prove valuable hosts for various DT gene-containing vectors that express the toxin intracellularly .

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interacts_with DB04868?

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P01308 action on H292 bronchial carcinoma cells as compared to normal bronchial epithelial cells . DB00030 may contribute to bronchial carcinoma due to P08069 activation by high local concentrations . Therefore , effects of insulin and P05019 on human bronchial carcinoma cells ( H292 ) and normal bronchial epithelium cells ( P02100 ) were studied . TGF-β was included since it also influences carcinoma progression . H292 and P02100 cells expressed both the insulin receptor and the P08069 ; in H292 cells an additional , shorter , splicing variant ( IR-A ) of the insulin receptor was present . P06213 expression was around four to five times higher in H292 than in P02100 cells at mRNA and protein levels . P01308 and TGF-β exerted contrary actions on proliferation and gene expression in H292 cells . Genes regulated by insulin , P05019 , and TGF-β were linked to inflammation , cell adhesion , muscle contraction and differentiation . P01308 and P05019 also suppressed DNA repair genes . EC(50) for insulin-induced proliferation was around 5 nM in H292 and around 30 nM P02100 cells . The EC(50) values for gene expression ranged from 9 to 90 nM in both cell types , dependent on the gene studied . In H292 cells , the proliferative response was much stronger if TGF-β was present . In P02100 cells this interaction of insulin and TGF-β was not observed , and changes in gene expression were mostly lower by at least 10-fold as compared to H292 . All in all , the insulin effects in H292 were generally much stronger than in P02100 cells and - with regard to proliferation - occurred at lower concentrations . Thus , insulin will hardly induce cancer from normal bronchial cells but may favour progression of pre-existing tumours . [ 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 ) ; 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 lymphocyte count , has been associated with systemic reactions and hepatitis in Caucasians , and the HLA-Cw8 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 ( MDR-1 ) gene codifying for glycoprotein P ( lower risk ) ; hyperbilirubinemia in patients exposed to atazanavir or indinavir carrying the P22309 *28 polymorphism ; peripheral neuropathy with nucleoside analogues associated with haplogroup T of the mitochondrial genome ( higher risk ) and with the Q30201 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 gene haplotypes associated with tenofovir-induced proximal tubulopathy , and the risk of pancreatitis in persons with mutations in the P13569 and SPINK-1 genes . Replacement of the transmembrane anchor in angiotensin I-converting enzyme ( P12821 ) with a glycosylphosphatidylinositol tail affects activation of the P30411 by P12821 inhibitors . To investigate further the relationship of angiotensin I-converting enzyme ( P12821 ) inhibitors to activation of the B(2) bradykinin ( BK ) receptor , we transfected Chinese hamster ovary cells to stably express the human receptor and either wild-type P12821 ( WT- P12821 ) , an P12821 construct with most of the cytosolic portion deleted ( Cyt-del- P12821 ) , or P12821 with a glycosylphosphatidylinositol ( P06744 ) anchor replacing the transmembrane and cytosolic domains ( P06744 - P12821 ) . BK or its P12821 -resistant analogue were the agonists . All activities ( arachidonic acid release and calcium mobilization ) were blocked by the B(2) antagonist DB06196 . B(2) was desensitized by repeated administration of BK but resensitized to agonist by P12821 inhibitors in the cells expressing both B(2) and either WT- P12821 or Cyt-del- P12821 . In P06744 - P12821 expressing cells , the B(2) receptor was still activated by the agonists , but P12821 inhibitors did not resensitize . Pretreatment with filipin returned the sensitivity to inhibitors . In immunocytochemistry , P06744 - P12821 showed patchy , uneven distribution on the plasma membrane that was restored by filipin . Thus , P12821 inhibitors were inactive as long as P06744 - P12821 was sequestered in cholesterol-rich membrane domains . WT- P12821 and B(2) receptor in Chinese hamster ovary cells co-immunoprecipitated with antibody to receptor , suggesting an interaction on the cell membrane . P12821 inhibitors augment BK effects on receptors indirectly only when enzyme and receptor molecules are sterically close , possibly forming a heterodimer . Inhibition of collagen-induced discoidin domain receptor 1 and 2 activation by imatinib , nilotinib and dasatinib . Imatinib , nilotinib and dasatinib are protein kinase inhibitors which target the tyrosine kinase activity of the Breakpoint Cluster Region-Abelson kinase ( P11274 - P00519 ) and are used to treat chronic myelogenous leukemia . Recently , using a chemical proteomics approach another tyrosine kinase , the collagen receptor Discoidin Domain Receptor1 ( Q08345 ) has also been identified as a potential target of these compounds . To further investigate the interaction of imatinib , nilotinib and dasatinib with Q08345 kinase we cloned and expressed human Q08345 and developed biochemical and cellular functional assays to assess their activity against Q08345 and the related receptor tyrosine kinase Discoidin Domain Receptor2 ( Q16832 ) . Our studies demonstrate that all 3 compounds are potent inhibitors of the kinase activity of both Q08345 and Q16832 . In order to investigate the question of selectivity among Q08345 , Q16832 and other tyrosine kinases we have aligned Q08345 and Q16832 protein sequences to other closely related members of the receptor tyrosine kinase family such as Muscle Specific Kinase ( O15146 ) , insulin receptor ( P06213 ) , Abelson kinase ( c- P00519 ) , and the stem cell factor receptor ( c- P10721 ) and have built homology models for the Q08345 and Q16832 kinase domains . In spite of high similarity among these kinases we show that there are differences within the DB00171 -phosphate binding loop ( P-loop ) , which could be exploited to obtain kinase selective compounds . Furthermore , the potent Q08345 and Q16832 inhibitory activity of imatinib , nilotinib and dasatinib may have therapeutic implications in a number of inflammatory , fibrotic and neoplastic diseases . P06870 protects cortical neurons against hypoxia/reoxygenation injury via the P27361 /2 pathway . Systemic or local delivery of human tissue kallikrein gene ( hTK ) has been shown to be an effective strategy to alleviate cerebral ischemia/reperfusion ( I/R ) injury , and tissue kallikrein ( TK ) administration can suppress glutamate- or acidosis-mediated neurotoxicity in vitro . In the present study , the role of TK in hypoxia/reoxygenation ( H/R ) induced neuronal cell death was investigated . We found that TK administration could remarkably alleviate H/R-induced neuronal injury by reduction of LDH release and promotion of neuron viability . The protective effects of TK could be counteracted by bradykinin B2 receptor ( P30411 ) antagonist HOE140 , which could suppress up-regulation of TK on the P29323 signal pathway under H/R condition . These results indicate that TK plays an important role in preventing neurons from H/R damage at least partially through the TK- P30411 - P27361 /2 pathway . Modulation of cytokine production and enhancement of cell viability by Q9NYK1 and Q9NR96 ligands during anthrax infection of macrophages . Inhalation of Bacillus anthracis , a bioterrorism agent , results in a high mortality rate despite appropriate antibiotic therapy . Macrophages appear to be a key factor in B. anthracis pathogenesis . The burst of pro-inflammatory cytokines from macrophages could be a major cause of death in anthrax . However , preactivation of Toll-like receptors ( TLRs ) could modify the host response . TLR ligands stimulate the release of activating cytokines but may also down-modulate the subsequent deleterious cytokine response to pathogens . We developed a cell culture model to measure macrophage responses to B. anthracis spores and bacilli . We found that germination from spores to bacilli produced a substantial stimulus for the secretion of the cytokines P05231 , P01375 , P22301 , and IL-12 p40 . Our studies showed that pretreatment of mouse macrophages with the Q9NR96 ligand DB05463 , or the Q9NYK1 ligands R-848 and IT-37 , results in a substantial decrease in the subsequent secretion of P05231 and P01375 in response to B. anthracis infection of macrophages . Furthermore , the Q9NYK1 and Q9NR96 ligands significantly decreased anthrax-induced cytotoxicity in the macrophages . These findings suggest that TLR ligands may contribute to the enhancement of innate immunity in B. anthracis infection by suppressing potentially deleterious pro-inflammatory cytokine responses and by improving macrophage viability . c-Src modulates estrogen-induced stress and apoptosis in estrogen-deprived breast cancer cells . The emergence of anti-estrogen resistance in breast cancer is an important clinical phenomenon affecting long-term survival in this disease . Identifying factors that convey cell survival in this setting may guide improvements in treatment . Estrogen ( E2 ) can induce apoptosis in breast cancer cells that have been selected for survival after E2 deprivation for long periods ( MCF-7:5C cells ) , but the mechanisms underlying E2-induced stress in this setting have not been elucidated . Here , we report that the c-Src kinase functions as a key adapter protein for the estrogen receptor ( ER , P03372 ) in its activation of stress responses induced by E2 in MCF-7:5C cells . E2 elevated phosphorylation of c-Src , which was blocked by 4-hydroxytamoxifen ( DB04468 ) , suggesting that E2 activated c-Src through the ER . We found that E2 activated the sensors of the unfolded protein response ( UPR ) , IRE1α ( O75460 ) and Q9NZJ5 kinase ( Q9NZJ5 ) , the latter of which phosphorylates eukaryotic translation initiation factor-2α ( eIF2α ) . E2 also dramatically increased reactive oxygen species production and upregulated expression of heme oxygenase P09601 ( P09601 ) , an indicator of oxidative stress , along with the central energy sensor kinase AMPK ( P54646 ) . Pharmacologic or RNA interference-mediated inhibition of c-Src abolished the phosphorylation of eIF2α and AMPK , blocked E2-induced ROS production , and inhibited E2-induced apoptosis . Together , our results establish that c-Src kinase mediates stresses generated by E2 in long-term E2-deprived cells that trigger apoptosis . This work offers a mechanistic rationale for a new approach in the treatment of endocrine-resistant breast cancer . Establishment and phenotypic characterization of human U937 cells with inducible P210 P11274 / P00519 expression reveals upregulation of P13688 ( CD66a ) . Chronic myeloid leukemia ( CML ) is characterized by the expression of the P210 P11274 / P00519 fusion protein . The molecular mechanisms behind this oncogene-mediated hematological disease are , however , not fully understood . Here , we describe the establishment and phenotypic characterization of U937 cells in which P210 P11274 / P00519 can be conditionally expressed using tetracycline . The induction of P11274 / P00519 in the obtained clones resulted in a rapid phosphorylation of the P42224 , P40763 and P42229 molecules , consistent with the findings in other model systems . Phenotypic characterization of the clones revealed that P11274 / P00519 induces a slight decrease in the proliferation and viability , without a marked effect on cell cycle distribution , the rate of apoptosis or on cellular differentiation , as judged by several cell surface markers and capacity to reduce nitro blue tetrazolium . Interestingly , P11274 / P00519 was found to upregulate the expression of carcinoembryonic-related antigen ( P06731 ) P62158 ( CD66a ) , which is a plasma membrane-linked glycoprotein belonging to the CEAs and involved in signal transduction and cellular adhesion . The expression of P13688 was reversible upon imatinib treatment in P11274 / P00519 -expressing U937 cells as well as in P11274 / P00519 -positive K562 cells . The established cell lines may prove useful in further modeling and dissection of P11274 / P00519 -induced leukemogenesis . Presynaptic serotonergic inhibition of GABAergic synaptic transmission in mechanically dissociated rat basolateral amygdala neurons . 1. The basolateral amygdala ( P00519 ) nuclei contribute to the process of anxiety . GABAergic transmission is critical in these nuclei and serotonergic inputs from dorsal raphe nuclei also significantly regulate GABA release . In mechanically dissociated rat P00519 neurons , spontaneous miniature inhibitory postsynaptic currents ( mIPSCs ) arising from attached GABAergic presynaptic nerve terminals were recorded with the nystatin-perforated patch method and pharmacological isolation . 2 . 5-HT reversibly reduced the GABAergic mIPSC frequency without affecting the mean amplitude . The serotonergic effect was mimicked by the P08908 specific agonist 8-OH DPAT ( 8-hydroxy-2-(di-n-propylamino)tetralin ) and blocked by the P08908 antagonist spiperone . 3 . The GTP-binding protein inhibitor N-ethylmaleimide removed the serotonergic inhibition of mIPSC frequency . In either K+-free or Ca2+-free external solution , 5-HT could inhibit mIPSC frequency . 4 . High K+ stimulation increased mIPSC frequency and 8-OH DPAT inhibited this increase even in the presence of Cd2+ . 5 . DB02587 , an activator of adenylyl cyclase ( AC ) , significantly increased synaptic GABA release frequency . Pretreatment with forskolin prevented the serotonergic inhibition of mIPSC frequency in both the standard and high K+ external solution . 6 . Ruthenium Red ( RR ) , an agent facilitating the secretory process in a Ca2+-independent manner , increased synaptic GABA release . 5-HT also suppressed RR-facilitated mIPSC frequency . 7 . We conclude that 5-HT inhibits GABAergic mIPSCs by inactivating the AC- DB02527 signal transduction pathway via a G-protein-coupled P08908 receptor and this intracellular pathway directly acts on the GABA-releasing process independent of K+ and Ca2+ channels in the presynaptic nerve terminals . Anti-allergic effects of nilotinib on mast cell-mediated anaphylaxis like reactions . DB04868 is a new orally bioavailable potent tyrosine kinase inhibitor that is used for the treatment of P11274 - P00519 -positive chronic myelogenous leukemia . However , its effect on mast cell-mediated anaphylactic reaction is still not known . The present study aimed to investigate the effect of nilotinib on the anaphylactic allergic reaction and study its possible mechanism(s) of action . DB04868 administration prevented systemic anaphylaxis in mice , mediated by compound 48/80 , in a dose- and time-dependent manner . Also , nilotinib significantly inhibited ( P < 0.05 ) allergic paw edema in rats . Furthermore , nilotinib significantly decreased ( P < 0.05 ) the IgE-mediated passive cutaneous anaphylaxis in a dose dependent manner . In addition , nilotinib dose-dependently reduced histamine release from the rat peritoneal mast cells activated either by compound 48/80 or by ovalbumin . Moreover , nilotinib attenuated the secretion of pro-inflammatory cytokine , tumor necrosis factor ( P01375 ) -α expression in the rat peritoneal mast cells . These findings provide evidence that nilotinib inhibits mast cell-derived immediate-type allergic reactions and so it could be a candidate as an anti-allergic agent . LSD and DOB : interaction with 5- Q13049 receptors to inhibit DB01221 receptor-mediated transmission in the rat prefrontal cortex . Both the phenethylamine hallucinogen (-)-1-2 , 5-dimethoxy-4-bromophenyl-2-aminopropane ( DOB ) , a selective serotonin 5- Q13049 ,2C receptor agonist , and the indoleamine hallucinogen DB04829 ( LSD , which binds to P08908 , 1B , 1D , 1E , 1F , 2A , 2C , 5 , 6 , 7 , dopamine D1 and D2 , and alpha1 and alpha2 adrenergic receptors ) , but not their non-hallucinogenic congeners , inhibited N-methyl-D-aspartate ( DB01221 ) -induced inward current and DB01221 receptor-mediated synaptic responses evoked by electrical stimulation of the forceps minor in pyramidal cells of the prefrontal cortical slices . The inhibitory effect of hallucinogens was mimicked by 5-HT in the presence of selective P08908 and 5- Q9H205 receptor antagonists . The inhibitory action of DOB , LSD and 5-HT on the DB01221 transmission was blocked by the 5- Q13049 receptor antagonists R-(+)-alpha- ( 2 , 3-dimethoxyphenil ) -1-[4-fluorophenylethyl]-4-piperidineme thanol ( M100907 ) and ketanserin . However , at low concentrations , when both LSD and DOB by themselves only partially depressed the DB01221 response , they blocked the inhibitory effect of 5-HT , suggesting a partial agonist action . Whereas N-(4-aminobutyl)-5-chloro-2-naphthalenesulphonamide ( W-7 , a calmodulin antagonist ) and N- [ 2- [ [ [ 3-(4'-chlorophenyl)- 2-propenyl ] methylamino ] methyl ] phenyl ] -N-(2-hydroxyethyl)-4'-methoxy-b enzenesulphonamide phosphate ( KN-93 , a Ca2+/ P62158 -KII inhibitor ) , but not the negative control 2-[N-4'methoxybenzenesulphonyl]amino-N-(4'-chlorophenyl)-2-propeny l-N -methylbenzylamine phosphate ( KN-92 ) , blocked the inhibitory action of LSD and DOB , the selective protein kinase C inhibitor chelerythrine was without any effect . We conclude that phenethylamine and indoleamine hallucinogens may exert their hallucinogenic effect by interacting with 5- Q13049 receptors via a Ca2+/ P62158 -KII-dependent signal transduction pathway as partial agonists and modulating the DB01221 receptors-mediated sensory , perceptual , affective and cognitive processes . Repression of human heat shock factor 1 activity at control temperature by phosphorylation . Human heat shock transcription factor 1 ( Q00613 ) is responsible for stress-induced transcription of heat shock protein genes . The activity of the Q00613 transcriptional activation domains is modulated by a separate regulatory domain , which confers repression at control temperature and heat inducibility . We show here that two specific proline-directed serine motifs are important for function of the regulatory domain : Mutation of these serines to alanine derepresses Q00613 activity at control temperature , and mutation to glutamic acid , mimicking a phosphorylated serine , results in normal repression at control temperature and normal heat shock inducibility . Tryptic mapping shows that these serines are the major phosphorylation sites of Q00613 at control temperature in vivo . Stimulation of the Raf/ P29323 pathway in vivo results in an increased level of phosphorylation of these major sites and the regulatory domain is an excellent substrate in vitro for the mitogen-activated MAPK/ P29323 . We conclude that phosphorylation of the regulatory domain of Q00613 decreases the activity of Q00613 at control temperature , and propose a mechanism for modification of Q00613 activity by growth control signals . Potentiation of DB04868 -mediated cell death in the context of the bone marrow microenvironment requires a promiscuous JAK inhibitor in CML . In this study , we show that conditioned media ( CM ) generated from bone marrow ( BM ) -derived mesenchymal stromal cells lead to P11274 - P00519 independent P40763 activation . Activation of P40763 is important not only for survival of CML cells but also for its protection against DB04868 ( NI ) , within the BM microenvironment . Reducing the expression of both O60674 and P29597 or utilizing a pan-JAK inhibitor blocked CM-mediated P40763 activation and sensitized CML cells to NI-mediated cell death . Finally , we demonstrate that in patient-derived primitive leukemic cells , co-cultured with BM stromal cells , inhibition of P11274 - P00519 and JAK activity was a successful strategy to potentiate their elimination . P43490 / P43490 /visfatin and cancer . P43490 / P43490 /visfatin is the rate-limiting enzyme that catalyzes the first step in NAD biosynthesis from nicotinamide and regulates growth , apoptosis and angiogenesis of mammalian cells . This enzyme was originally cloned as a putative cytokine shown to enhance the B cell precursor maturation in the presence of P13232 and stem cell factor . A number of cancers have increased expression of P43490 / P43490 /visfatin , which regulates a variety of different signaling pathways such as PI3K/Akt , P27361 /2 and P40763 . FK866/APO866 and CHS828/ DB05217 are two known inhibitors of P43490 / P43490 /visfatin and have been evaluated as anticancer agents in the clinic . This review will focus on its role in carcinogenesis and cancer progression and its inhibitors as therapeutic target for cancer treatment . Association between severe toxicity of nilotinib and P22309 polymorphisms in Japanese patients with chronic myelogenous leukemia . BACKGROUND : DB04868 is a P11274 - P00519 kinase inhibitor approved for the treatment of Philadelphia chromosome-positive chronic myelogenous leukemia ( CML ) . The P22309 ( P22309 ) polymorphism P22309 *28 ( *28 ) /*28 has been linked to an increased risk of hyperbilirubinemia in patients with CML who receive nilotinib . Beside *28 , P22309 *6 ( *6 ) is another important variant allele in Japanese patients because it is associated with adverse events of irinotecan , metabolized by P22309 . We retrospectively investigated the association between severe toxicity of nilotinib and P22309 polymorphisms ( *6 and*28 ) in Japanese patients with CML . PATIENTS AND METHODS : Eight patients with cytogenetically confirmed CML who were receiving nilotinib were studied to explore the association of P22309 polymorphisms with severe nilotinib-related toxicity . Genotyping analyses were determined for *6 and *28 . RESULTS : All 3 patients with the *6/*6 or *6/*28 genotype had severe toxicity , including QT interval prolongation ( grade 3 ) , elevated lipase levels ( grade 3 ) plus hyperbilirubinemia ( grade 2 ) , and anemia ( grade 3 ) plus hepatic cyst hemorrhage ( grade 2 ) in 1 patient each . Among the 5 patients with the *6/*1 or *1/*1 genotype , 1 had elevated lipase levels ( grade 3 ) and another had severe pain in the lower extremities ( grade 3 ) . CONCLUSION : These findings suggest that P22309 polymorphisms are important determinants of severe toxicity of nilotinib in Japanese patients . DB06016 , an orally active D2/D3 receptor antagonist , for the potential treatment of schizophrenia , bipolar mania and depression . DB06016 ( RGH-188 ) , which is being codeveloped by Gedeon Richter Ltd , Forest Laboratories Inc and Mitsubishi Tanabe Pharma Corp , is a novel putative antipsychotic drug that exerts partial agonism at dopamine D2/D3 receptors , with preferential binding to D3 receptors , and partial agonism at serotonin P08908 receptors . Its activity at D2/D3 receptors may be lower than that of the prototype partial agonist aripiprazole . The antipsychotic activity of cariprazine was demonstrated in animal models , and data also suggest that the propensity for extrapyramidal side effects is low and that the drug may have procognitive properties . DB06016 is rapidly absorbed , with high oral bioavailability and a long plasma elimination t1/2 . DB06016 is in phase III clinical trials in patients with schizophrenia and in patients with bipolar disorder . Data from phase II trials in patients with schizophrenia and bipolar mania indicate that the drug has antipsychotic and antimanic properties that are superior to placebo . With its unique receptor affinity profile , cariprazine may represent a potential enrichment of the therapeutic armamentarium for schizophrenia and affective disorders . Its activity against the cognitive deficits associated with schizophrenia has to be carefully investigated . DB03758 activates heat shock protein expression and cardioprotection in neonatal rat cardiomyocytes . Heat shock proteins ( HSPs ) constitute an endogenous cellular defense mechanism against environmental stresses . In the past few years , studies have shown that overexpression of HSPs can protect cardiac myocytes against ischemia-reperfusion injury . In an attempt to increase the HSPs in cardiac tissue , we used the compound radicicol that activates HSP expression by binding to the P08238 kDa ( HSP90 ) . HSP90 is the main component of the cytosolic molecular chaperone complex , which has been implicated in the regulation of the heat shock factor 1 ( Q00613 ) . Q00613 is responsible for the transcriptional activation of the heat shock genes . In the present study , we show that radicicol induces HSP expression in neonatal rat cardiomyocytes , and this increase in HSPs confers cardioprotection to these cardiomyocytes . We also show that radicicol induction of the HSP and cardioprotection is dependent on the inhibition of HSP90 in cardiomyocytes . These results indicate that modulation of the active HSP90 protein level plays an important role in cardioprotection . Therefore , compounds , such as radicicol and its possible derivatives that inhibit the function of HSP90 in the cell may represent potentially useful cardioprotective agents . The Q9NZJ5 -eIF2α phosphorylation arm is a pro-survival pathway of P11274 - P00519 signaling and confers resistance to imatinib treatment in chronic myeloid leukemia cells . Activation of adaptive mechanisms plays a crucial role in cancer progression and drug resistance by allowing cell survival under stressful conditions . Therefore , inhibition of the adaptive response is considered as a prospective therapeutic strategy . The Q9NZJ5 -eIF2α phosphorylation pathway is an important arm of the unfolded protein response ( UPR ) , which is induced under conditions of endoplasmic reticulum ( ER ) stress . Our previous work showed that ER stress is induced in chronic myeloid leukemia ( CML ) cells . Herein , we demonstrate that the Q9NZJ5 -eIF2α phosphorylation pathway is upregulated in CML cell lines and P28906 + cells from CML patients and is associated with CML progression and imatinib resistance . We also show that induction of apoptosis by imatinib results in the downregulation of the Q9NZJ5 -eIF2α phosphorylation arm . Furthermore , we demonstrate that inactivation of the Q9NZJ5 -eIF2α phosphorylation arm decreases the clonogenic and proliferative capacities of CML cells and sensitizes them to death by imatinib . These findings provide evidence for a pro-survival role of Q9NZJ5 -eIF2α phosphorylation arm that contributes to CML progression and development of imatinib resistance . Thus , the Q9NZJ5 -eIF2α phosphorylation arm may represent a suitable target for therapeutic intervention for CML disease . DB04868 and MEK inhibitors induce synthetic lethality through paradoxical activation of RAF in drug-resistant chronic myeloid leukemia . We show that imatinib , nilotinib , and dasatinib possess weak off-target activity against RAF and , therefore , drive paradoxical activation of P15056 and CRAF in a DB01367 -dependent manner . Critically , because DB01367 is activated by P11274 - P00519 , in drug-resistant chronic myeloid leukemia ( CML ) cells , DB01367 activity persists in the presence of these drugs , driving paradoxical activation of P15056 , CRAF , MEK , and P29323 , and leading to an unexpected dependency on the pathway . Consequently , nilotinib synergizes with MEK inhibitors to kill drug-resistant CML cells and block tumor growth in mice . Thus , we show that imatinib , nilotinib , and dasatinib drive paradoxical RAF/MEK/ P29323 pathway activation and have uncovered a synthetic lethal interaction that can be used to kill drug-resistant CML cells in vitro and in vivo . DB01645 stimulates duodenal HCO(3)(-) secretion through PI3K pathway in mice . DB01645 has been proposed as a promising pharmacotherapeutic for cystic fibrosis . We recently found that genistein stimulates murine duodenal HCO(3)(-) secretion through cystic fibrosis transmembrane conductance regulator ( P13569 ) . The aim of the present study was to determine the intracellular signal pathways involved in genistein-stimulated duodenal HCO(3)(-) secretion . Murine duodenal mucosal HCO(3)(-) secretion was examined in vitro in Ussing chambers by the pH-stat technique . The results showed that neither DB02527 -dependent signal pathway inhibitors MDL-12330A and KT-5720 , nor cGMP signal pathway inhibitors NS2028 and KT5823 , nor calcium signal pathway inhibitors verapamil and W-13 , altered genistein-stimulated duodenal HCO(3)(-) secretion . In calcium-free solution , genistein-stimulated duodenal HCO(3)(-) secretion was not altered either . Vanadate , an inhibitor of protein tyrosine phosphatase , only partially inhibited genistein-stimulated duodenal HCO(3)(-) secretion . However , both wortmannin and LY294002 , two structurally and mechanistically distinct phosphatidylinositol 3-kinase ( PI3K ) inhibitors , markedly inhibited genistein-stimulated duodenal HCO(3)(-) secretion . DB01645 increased duodenal mucosal PI3K activity and induced the phosphorylation of Akt , a signaling molecule downstream of PI3K , which was again inhibited by wortmannin . P03372 antagonist , ICI182,780 , also markedly inhibited genistein-stimulated duodenal HCO(3)(-) secretion and genistein-induced PI3K activity increase in duodenal mucosa . These results demonstrate that genistein stimulates duodenal HCO(3)(-) secretion mainly through estrogen receptor and PI3K-dependent pathway . These findings contribute to the understanding of the molecular mechanism of genistein-induced anion secretion and further pharmacotherapeutic development and use of genistein or related substances in the treatment of diseases of epithelial tissues .

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interacts_with DB01151?

multiple_choice

8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . Comparison of the novel antipsychotic ziprasidone with clozapine and olanzapine : inhibition of dorsal raphe cell firing and the role of P08908 receptor activation . Ziprasidone is a novel antipsychotic agent which binds with high affinity to P08908 receptors ( Ki = 3.4 nM ) , in addition to P28221 , 5-HT2 , and D2 sites . While it is an antagonist at these latter receptors , ziprasidone behaves as a P08908 agonist in vitro in adenylate cyclase measurements . The goal of the present study was to examine the P08908 properties of ziprasidone in vivo using as a marker of central P08908 activity the inhibition of firing of serotonin-containing neurons in the dorsal raphe nucleus . In anesthetized rats , ziprasidone dose-dependently slowed raphe unit activity ( ED50 = 300 micrograms/kg i.v. ) as did the atypical antipsychotics clozapine ( ED50 = 250 micrograms/kg i.v. ) and olanzapine ( ED50 = 1000 micrograms/kg i.v. ) . Pretreatment with the P08908 antagonist WAY-100,635 ( 10 micrograms/kg i.v. ) prevented the ziprasidone-induced inhibition ; the same dose of WAY-100,635 had little effect on the inhibition produced by clozapine and olanzapine . Because all three agents also bind to alpha 1 receptors , antagonists of which inhibit serotonin neuronal firing , this aspect of their pharmacology was assessed with desipramine ( DB01151 ) , a NE re-uptake blocker previously shown to reverse the effects of alpha 1 antagonists on raphe unit activity . DB01151 ( 5 mg/kg i.v. ) failed to reverse the inhibitory effect of ziprasidone but produced nearly complete reversal of that of clozapine and olanzapine . These profiles suggest a mechanism of action for each agent , P08908 agonism for ziprasidone and alpha 1 antagonism for clozapine and olanzapine . The P08908 agonist activity reported here clearly distinguishes ziprasidone from currently available antipsychotic agents and suggests that this property may play a significant role in its pharmacologic actions . An acetylcholinesterase inhibitor , eserine , induces long-term depression at P07451 - P00915 synapses in the hippocampus of adult rats . Studies in humans and rodents support a role for muscarinic ACh receptor ( mAChR ) and nicotinic AChR in learning and memory , and both regulate hippocampal synaptic plasticity using complex and often times opposing mechanisms . P22303 ( P22303 ) inhibitors are commonly prescribed to enhance cholinergic signaling in Alzheimer 's disease in hopes of rescuing cognitive function , caused , in part , by degeneration of cholinergic innervation to the hippocampus and cortex . Unfortunately , therapeutic efficacy is moderate and inconsistent , perhaps due to unanticipated mechanisms . M1 mAChRs bidirectionally control synaptic strength at P07451 - P00915 synapses ; weak pharmacological activation using carbachol ( CCh ) facilitates potentiation , whereas strong agonism induces muscarinic long-term depression ( mLTD ) via an P29323 -dependent mechanism . Here , we tested the prediction that accumulation of extracellular ACh via inhibition of P22303 is sufficient to induce LTD at P07451 - P00915 synapses in hippocampal slices from adult rats . Although P22303 inhibition with eserine induces LTD , it unexpectedly does not share properties with mLTD induced by CCh , as reported previously . DB00981 -LTD was prevented by the M3 mAChR-preferring antagonist 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide ( 4-DAMP ) , and pharmacological inhibition of MEK was completely ineffective . Additionally , pharmacological inhibition of p38 MAPK prevents mLTD but has no effect on eserine-LTD . Finally , long-term expression of eserine-LTD is partially dependent on a decrease in presynaptic release probability , likely caused by tonic activation of mAChRs by the sustained increase in extracellular ACh . Thus these findings extend current literature by showing that pharmacological P22303 inhibition causes a prolonged decrease in presynaptic glutamate release at P07451 - P00915 synapses , in addition to inducing a likely postsynaptic form of LTD . Vascular endothelial growth factor signaling is required for the behavioral actions of antidepressant treatment : pharmacological and cellular characterization . This study extends earlier work on the role of vascular endothelial growth factor ( P15692 ) in the actions of antidepressant treatment in two key areas . First , by determining the requirement for P15692 in the actions of a 5-HT selective reuptake inhibitor ( SSRI ) , fluoxetine in behavioral models of depression/antidepressant response ; and second , by examining the role of the P08908 receptor subtype in the regulation of P15692 , and the cellular localization of antidepressant regulation of P15692 expression . The results show that pharmacological inhibition of P15692 receptor signaling blocks the behavioral actions of fluoxetine in rats subjected to chronic unpredictable stress . Infusions of SU5416 or SU1498 , two structurally dissimilar inhibitors of P15692 -Flk-1 receptor signaling , block the antidepressant effects of fluoxetine on sucrose preference , immobility in the forced swim test , and latency to feed in the novelty suppressed feeding paradigm . We also show that activation of P08908 receptors is sufficient to induce P15692 expression and that a P08908 antagonist blocks both the increase in P15692 and behavioral effects induced by fluoxetine . Finally , double labeling studies show that chronic fluoxetine administration increases P15692 expression in both neurons and endothelial cells in the hippocampus . Taken together these studies show that P15692 is necessary for the behavioral effects of the SSRI fluoxetine , as well as norepinephrine selective reuptake inhibitor , and that these effects may be mediated by P08908 receptors located on neurons and endothelial cells . The bile acid receptor Q96RI1 is a modulator of intestinal innate immunity . The farnesoid X receptor ( Q96RI1 ) is a bile acid-regulated nuclear receptor expressed in enterohepatic tissues . In this study we investigated whether Q96RI1 is expressed by cells of innate immunity and regulates inflammation in animal models of colitis . Acute ( 7 days ) and chronic ( 8 wk ) colitis were induced in wild-type and Q96RI1 (-/-) mice by intrarectal administration of trinitrobenzensulfonic acid or by 7-day administration of 5 % dextran sulfate in drinking water . The results of this experiment demonstrate that Q96RI1 is expressed by and exerts counterregulatory effects on cells of innate immunity . Exposure of LPS-activated macrophages to 6-ethyl chenodeoxycholic acid ( 6E- DB06777 ; INT-747 ) a synthetic Q96RI1 ligand , results in a reciprocal regulation of NF-kappaB dependent-genes ( P01375 , IL-1beta , P05231 , P23219 , P35354 , and P35228 ) and induction of Q15466 , a Q96RI1 -regulated gene . Q96RI1 activation stabilizes the nuclear corepressor NCoR on the NF-kappaB responsive element on the IL-1beta promoter . Colon inflammation in Crohn 's disease patients and in rodent models of colitis is associated with a reduced expression of Q96RI1 mRNA . Using two rodent models of colon inflammation , we show that progression of these immune-mediated disorders is exacerbated in Q96RI1 (-/-) mice ( p < 0.01 ) . In vivo treatment with INT-747 attenuates organ injury and immune cell activation . Q96RI1 activation increased the colon expression of P51161 , Q96RI1 , and Q15466 while reducing IL-1beta , P60568 , P05231 , P01375 , and P01579 mRNA expression and attenuating disease severity . In aggregate , these findings provide evidence that Q96RI1 is an essential component of a network of nuclear receptors that regulate intestinal innate immunity and homeostasis . Stimulation of cloned human serotonin P28221 beta receptor sites in stably transfected P13671 glial cells promotes cell growth . The involvement of serotonin P28221 beta receptor sites was investigated in the growth of rat P13671 glial cells permanently transfected with a gene encoding a human P28221 beta receptor . The 5-HT receptor identity of control and transfected P13671 glial/ P28221 beta cells was determined by reverse transcription-polymerase chain reaction using primers specific for rat P08908 , rat P28222 , rat P28221 alpha , human P28221 beta , and rat 5- Q13049 receptor genes . Constitutive mRNA for 5- Q13049 receptors was present in control and transfected P13671 glial/ P28221 beta cells , whereas mRNA for P28221 beta receptor sites was only present in the transfected P13671 glial/ P28221 beta cell line . 5-HT inhibited forskolin-stimulated cyclic AMP formation and promoted cell growth , in contrast to the absence of any measurable effect in pcDNA3 plasmid-transfected and nontransfected P13671 glial cells . The 5-HT effects could be mimicked by sumatriptan ( EC50 = 44-76 nM ) and were totally and partially blocked by methiothepin ( IC50 = 9 nM ) and GR 127,935 ( 2'-methyl-4'-(5-methyl[1,2,4]oxadiazol-3-yl)-biphenyl-4-carbox yli c acid [4-methoxy-3-(4-methylpiperazin-1-yl)phenyl]amide ; IC50 = 97 pM ) , respectively . No effect on cell growth was measured with the 5-HT2 receptor agonist DOI [ 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane ; 10 microM ] , suggesting that 5- Q13049 receptors are not involved in the 5-HT-stimulated P13671 glial/ P28221 beta cell growth . Dibutyryl-cyclic AMP ( 0.3 mM ) -treated cultures did not show sumatriptan-promoted cell growth , indicating an inhibitory role for cyclic AMP in the cell growth mediated by P28221 beta receptor sites. ( ABSTRACT TRUNCATED AT 250 WORDS ) Q08462 selectively couples to E prostanoid type 2 receptors , whereas adenylyl cyclase 3 is not receptor-regulated in airway smooth muscle . Adenylyl cyclases ( ACs ) are important regulators of airway smooth muscle function , because β-adrenergic receptor ( βAR ) agonists stimulate AC activity and DB02527 production . We have previously shown in a number of cell types that AC6 selectively couples to βAR and these proteins are coexpressed in lipid rafts . We overexpressed AC2 , O60266 , and AC6 in mouse bronchial smooth muscle cells ( mBSMCs ) and human embryonic kidney ( P29320 ) -293 cells by using recombinant adenoviruses and assessed their localization and regulation by various G protein-coupled receptors ( GPCRs ) . O60266 and AC6 were expressed primarily in caveolin-rich fractions , whereas AC2 expression was excluded from these domains . AC6 expression enhanced DB02527 production in response to isoproterenol but did not increase responses to butaprost , reflecting the colocalization of AC6 with β(2)AR but not E prostanoid type 2 receptor ( EP(2)R ) in lipid raft fractions . AC2 expression enhanced butaprost-stimulated DB02527 production but had no effect on the β(2)AR-mediated response . O60266 did not couple to any GPCR tested . DB02587 -induced arborization of mBSMCs was assessed as a functional readout of DB02527 signaling . Arborization was enhanced by overexpression of AC6 and O60266 , but AC2 had no effect . GPCR-stimulated arborization mirrored the selective coupling observed for DB02527 production . With the addition of the phosphodiesterase 4 ( DB05876 ) inhibitor rolipram AC2 accelerated forskolin-stimulated arborization . Thus , AC2 selectively couples to EP(2)R , but signals from this complex are limited by DB05876 activity . O60266 does not seem to couple to GPCR in either mBSMCs or P29320 -293 cells , so it probably exists in a distinct signaling domain in these cells . A phase II study of DB05434 ( thrombospondin-1 analog ) for the treatment of metastatic melanoma . OBJECTIVES : Thrombospondins are natural inhibitors of angiogenesis , tumor metastases , and tumor growth ( melanoma ) . DB05434 is a synthetic analog of thrombospondin-1 , well tolerated in phase I studies . We conducted a phase II trial evaluating the clinical efficacy of DB05434 and its effects on biomarkers of angiogenesis and immunity in patients with metastatic melanoma ( MM ) . PATIENTS AND METHODS : A 2-stage phase II clinical trial was conducted to assess the clinical efficacy , safety , and pharmacodynamic effects ( angiogenesis and immunity ) of DB05434 in patients with stage IV melanoma . The primary endpoint was 18-week treatment failure rate . Patients self-administered 100 mg of DB05434 subcutaneously twice daily . Blood samples were collected at baseline and every 3 weeks while on therapy . Eligible patients demonstrated measurable disease , good performance status and no evidence of intracranial metastases . Correlative laboratory studies evaluated biomarkers of angiogenesis and immunity . RESULTS : Twenty-one patients were enrolled . Most patients were stage M1c ( 71 % ) and all had prior therapy for MM . Only 3 of the first 20 patients enrolled were progression free and on treatment at 18 weeks resulting in early termination of the study . Decreases in peripheral blood P15692 levels and P49767 levels , and CD146 and P28906 /133 counts relative to pretreatment were detected . Limited changes in antitumor T cell immunity were observed . CONCLUSIONS : DB05434 therapy administered at 100 mg twice/day in patients with MM did not demonstrate definite clinical efficacy . Further dose escalation or combination with cytotoxic therapy may be more effective therapeutically . Tandospirone activates neuroendocrine and P29323 ( Q96HU1 kinase ) signaling pathways specifically through P08908 receptor mechanisms in vivo . Tandospirone , an azapirone , is a selective serotonin(1A) ( 5-HT(1A) ) receptor agonist . The effects of tandospirone on plasma hormones and on mitogen-activated protein ( Q96HU1 ) kinase activity in the brain of male rats were studied . Tandospirone produced a time- and dose-dependent increase in plasma levels of oxytocin , adrenocorticotropin ( DB01285 ) , corticosterone , and prolactin . The minimal dose of tandospirone that led to a significant elevation of plasma oxytocin , DB01285 , and prolactin levels was 1.0 mg/kg ( s.c. ) , while the minimal dose for corticosterone release was 3.0 mg/kg ( s.c. ) . The ED(50) of tandospirone was 1.3 mg/kg for oxytocin , 1.2 mg/kg for DB01285 , 3.0 mg/kg for corticosterone , and 0.24 mg/kg for prolactin . Pretreatment with the specific 5-HT(1A) receptor antagonist WAY 100,635 ( 0.3 mg/kg , s.c. ) completely blocked the effects of tandospirone on plasma levels of oxytocin , DB01285 , and corticosterone but shifted the dose-response curve for prolactin to the right . Tandospirone injection ( 10 mg/kg , s.c. ) stimulated the Q96HU1 kinase signaling cascade , specifically the phosphorylation of Q8NFH3 /44 extracellular signal-regulated kinase ( P29323 ) . Western blot analysis revealed a significant increase in phosphorylated P29323 ( p- P29323 ) levels in the hypothalamic paraventricular nucleus ( PVN ) as well as the dorsal raphe nucleus 5 min following tandospirone injection . These increases were blocked by pretreatment with WAY 100,635 ( 0.3 mg/kg ) . The results are the first evidence that systemic 5-HT(1A) receptor agonist administration produces a rapid increase in p- P29323 levels in vivo , providing further insight into the signaling mechanisms of the 5-HT(1A) receptor . P35372 phosphorylation , desensitization , and ligand efficacy . Mu opioid receptors are subject to phosphorylation and desensitization through actions of at least two distinct biochemical pathways : agonist-dependent mu receptor phosphorylation and desensitization induced by a biochemically distinct second pathway dependent on protein kinase C activation ( 1 ) . To better understand the nature of the agonist-induced mu receptor phosphorylation events , we have investigated the effects of a variety of opiate ligands of varying potencies and intrinsic activities on mu receptor phosphorylation and desensitization . Exposure to the potent full agonists sufentanil , dihydroetorphine , etorphine , etonitazine , and [ D-Ala2 , MePhe4 , Glyol5 ] enkephalin ( DAMGO ) led to strong receptor phosphorylation , while methadone , l-alpha-acetylmethadone ( DB01227 ) , morphine , meperidine , DADL , beta-endorphin(1-31) , enkephalins , and dynorphin A(1-17) produced intermediate effects . The partial agonist buprenorphine minimally enhanced receptor phosphorylation while antagonists failed to alter phosphorylation . DB00921 and full antagonists each antagonized the enhanced mu receptor phosphorylation induced by morphine or DAMGO . The rank order of opiate ligand efficacies in producing mu receptor-mediated functional desensitization generally paralleled their rank order of efficacies in producing receptor phosphorylation . Interestingly , the desensitization and phosphorylation mediated by methadone and DB01227 were disproportionate to their efficacies in two distinct test systems . This generally good fit between the efficacies of opiates in mu receptor activation , phosphorylation , and desensitization supports the idea that activated receptor/agonist/G-protein complexes and/or receptor conformational changes induced by agonists are required for agonist-induced mu receptor phosphorylation . Data for methadone and DB01227 suggest possible contribution from their enhanced desensitizing abilities to their therapeutic efficacies . Functional characterization of a novel serotonin receptor ( 5-HTap2 ) expressed in the CNS of Aplysia californica . Serotonin has been shown to be a neuromodulator in the Aplysia californica CNS . The diversity of serotonin actions is due to the existence of several different receptor subtypes . In this study we report the cloning of a full-length cDNA , coding for a novel serotonin receptor ( 5-HTap2 ) . The receptor protein bears the characteristics of G protein-coupled receptors . It shares 68 % and 34 % of its amino acid sequence identity with the 5-HTlym receptor from Lymnaea stagnalis and the mammalian P08908 receptor , respectively . When transfected in P29320 293 cells , 5-HTap2 was negatively coupled to adenylate cyclase . Ligand binding analysis indicated that the order of potencies of various drugs for the inhibition of [3H]LSD binding was : methiothepin > metergoline > 5-CT > PAPP > 5-HT > ketanserin > NAN-190 > 8-OH-DPAT > clozapine . RT-PCR amplification of RNA isolated from different tissues indicated that this receptor is expressed in the CNS and in bag cells . The expression of 5-HTap2 restricted to the CNS suggests an important role for this receptor in the modulation of neuronal functions in Aplysia . Moreover , the high expression of 5-HTap2 in the bag cells , associated with its pharmacological profile , suggests that this receptor may be implicated in modulating the afterdischarge during the egg-laying behavior . Characterization of a novel Q13639 receptor antagonist of the azabicycloalkyl benzimidazolone class : DAU 6285 . Three chemical classes of serotonin Q13639 receptor agonists have been identified so far : 5-substituted indoles ( e.g. 5-HT ) , benzamides ( e.g. renzapride ) and benzimidazolones ( e.g. BIMU 8 ) . In a search for Q13639 receptor antagonists , we have discovered that the benzimidazolone derivative DAU 6285 ( for structure see text ) , is 3-5 times more potent than tropisetron in blocking 5-HT , renzapride and BIMU 8 induced stimulation of adenylate cyclase activity in mouse embryo colliculi neurons . Schild plot analysis yielded Ki values of 220 , 181 and 255 nmol/l , respectively . In addition , DAU 6285 showed poor activity as a 5- Q9H205 receptor ligand with respect to tropisetron , as demonstrated by in vitro binding studies ( Ki , 322 vs 2.8 nmol/l ) and by its antagonistic activity in the Bezold-Jarisch reflex test ( ID50 , 231 vs 0.5 micrograms/kg , i.v. ) . No significant binding ( Ki greater than 10 mumol/l ) of DAU 6285 to serotonergic P08908 , P28222 , P28335 , P28221 , and 5-HT2 receptors as well as to adrenergic alpha 1 , alpha 2 , dopaminergic D1 , D2 or muscarinic M1-M3 receptor subtypes was found . The data indicate that DAU 6285 has a somewhat higher affinity than tropisetron for Q13639 receptors , a property confirmed in functional tests , and much lower affinity than tropisetron for 5- Q9H205 receptors . The compound represents a new interesting tool for investigating the pharmacological and physiological properties of Q13639 receptors . Autoantibodies against four kinds of neurotransmitter receptors in psychiatric disorders . There is a hypothesis that autoimmune abnormalities in neurotransmitter receptors might cause some psychiatric disorders . Using a sensitive radioligand assay , we detected serum autoantibodies to recombinant human muscarinic cholinergic receptor 1 ( P11229 , 34.4 % ) , mu-opioid receptor ( P35372 , 13.1 % ) , P08908 ( P08908 , 7.4 % ) , and dopamine receptor D2 ( P14416 , 4.9 % ) in 122 psychiatric patients . Positive antibodies to P11229 were found in 34.1 % , 34.9 % , 33.3 % , and 9.1 % of patients with schizophrenic disorders ( n=44 ) , mood disorders ( n=63 ) , other psychiatric disorders ( n=15 ) and autoimmune diseases ( n=33 ) , respectively . All three patients with neuroleptic maliganant syndrome had high activities of autoantibodies to P11229 , P35372 , and/or P08908 . Our data suggest that autoimmunity to neurotransmitter receptors might be associated with the induction of psychiatric symptoms and have some relation to neuroleptic malignant syndrome . Structure-activity relationships in Q9NYK1 agonistic 1H-imidazo[4,5-c]pyridines . Engagement of Q9NYK1 in plasmacytoid dendritic cells leads to the induction of IFN-α/β which plays essential functions in the control of adaptive immunity . We had previously examined structure-activity relationships ( SAR ) in Q9NYK1 /8-agonistic imidazoquinolines with a focus on substituents at the N(1) , C(2) , N(3) and N(4) positions , and we now report SAR on 1H-imidazo[4,5-c]pyridines . 1-Benzyl-2-butyl-1H-imidazo[4,5-c]pyridin-4-amine was found to be a pure Q9NYK1 -agonist with negligible activity on Q9NR97 . Increase in potency was observed in N(6)-substituted analogues , especially in those compounds with electron-rich substituents . Direct aryl-aryl connections at P13671 abrogated activity , but Q9NYK1 agonism was reinstated in 6-benzyl and 6-phenethyl analogues . Consistent with the pure Q9NYK1 -agonistic behavior , prominent IFN-α induction in human PBMCs was observed with minimal proinflammatory cytokine induction . A benzologue of imidazoquinoline was also synthesized which showed substantial improvements in potency over the parent imidazopyridine . Distinct differences in N(6)-substituted analogues were observed with respect to IFN-α induction in human PBMCs on the one hand , and Q07108 upregulation in lymphocytic subsets , on the other . DB04860 , an agonist of Q9NYK1 , reduces plasma virus concentration in chronic hepatitis C infection . Immune-based therapy is the mainstay treatment for chronic hepatitis C virus ( HCV ) infection but causes multiple side effects and achieves durable viral clearance in only approximately 50 % of patients . Most new investigational anti-HCV compounds are direct-acting antivirals for which durability of response and risk of viral mutations and resistance are not yet known . Therefore , continuing discovery and development of new immune-based treatments is desirable . Toll-like receptors ( TLRs ) are pathogen recognition receptors that initiate the innate immune response . The responsiveness of HCV or other ongoing chronic systemic infections to treatment with a selective TLR agonist has not been reported . DB04860 is a selective agonist of Q9NYK1 . In a proof-of-concept study , we found that once-daily 7-day treatment with intravenous isatoribine 800 mg caused a significant ( P = .001 ) reduction of plasma HCV RNA ( mean , -0.76 ; range , -2.85 to +0.21 log(10) units ) in otherwise untreated patients ( n = 12 ) who were chronically infected with HCV . Viral load reduction occurred in patients infected with genotype 1 as well as non-genotype 1 HCV . The reduction of viral load was correlated with induction of markers of a heightened immune antiviral state , including 2'- , 5'- oligoadenylate synthetase levels in whole blood . This treatment was well tolerated , with a low frequency of mild to moderate adverse events . In conclusion , systemic administration of the selective Q9NYK1 agonist isatoribine resulted in dose-dependent changes in immunologic biomarkers and a statistically significant antiviral effect with relatively few and mild side effects . DB02533 treatment ameliorates inflammatory responses and memory impairment induced by amyloid-beta 25-35 injection in rats . Alzheimer disease ( AD ) is a neurodegenerative disorder caused by accumulation of the amyloid-beta peptide ( Aβ ) in neuritic plaques . Its neurotoxic mechanisms are associated with inflammatory responses and nitrosative stress generation that promote expression of inducible nitric oxide synthase ( P35228 ) and increased nitric oxide causing neuronal death and memory impairment . Studies suggest that treatment with anti-inflammatory and anti-oxidant agents decreases the risk of developing AD . DB02533 ( AG ) is an P35228 inhibitor with anti-inflammatory and anti-oxidant effects . In this study , we evaluated the effects of systemic administration of AG ( 100 mg/kg/day for 4 days ) on spatial memory and inflammatory responses induced by an injection of Aβ(25-35) [ 100 μM ] into the temporal cortex ( TCx ) of rats . A significant improvement of spatial memory was evident in the Aβ(25-35)-treated group at day 30 post-injection subjected to AG treatment ; this effect was correlated with decreases in reactive gliosis , IL-1β , P01375 -α , and nitrite levels , as well as a reduction in neurodegeneration in the TCx and hippocampus ( Hp ) . These results suggest that AG treatment inhibited glia activation and cytokine release , which may help to counteract neurodegenerative events induced by the toxicity of Aβ . [ Molecular mechanism of cardiovascular damage induced by aldosterone ] . Although the pro-inflammatory and pro-fibrotic actions of aldosterone on the vasculature have been reported , the effects and molecular mechanisms of aldosterone on endothelial function are yet to be determined . We investigated how aldosterone regulates endothelial nitric oxide synthase ( P29474 ) function in human umbilical vein endothelial cells ( HUVECs ) . HUVECs were incubated for 16 hrs with 10(-7) mol/l of aldosterone . The concentration of reactive oxygen species ( ROS ) was estimated by measuring DCF chemiluminescence . Signal transduction was estimated by Western immunoblots . Realtime RT-PCR was performed to measure expression of transcripts of endogenous GTP cyclohydrolase-1 ( P30793 ) and components of NAD(P)H oxidase . In order to eliminate the possible effect of the glucocorticoid receptor ( GR ) , and to emphasize the role of mineralocorticoid receptor ( MR ) , we used GR siRNA and knocked down GR expression in several experiments . NO output was estimated by intracellular cGMP concentration . ROS production increased significantly in aldosterone-treated HUVEC , but was abolished by pre-treatment with eplerenone . Transcripts of p47(phox) were increased by aldosterone treatment . Vascular endothelial growth factor ( P15692 ) -induced P29474 DB00133 1177 but not Akt DB00133 473 phosphorylation levels were reduced significantly by pretreatment with aldosterone . Pretreatment with either eplerenone or okadaic acid restored phosphorylation levels of P29474 DB00133 1177 in aldosterone-treated cells , suggesting that protein phosphatase ( PP ) 2A was upregulated by aldosterone via MR . The decrease in NO output caused by aldosterone pretreatment was reversed significantly by either DB00360 ( BH(4) ) , P30793 overexpression , or p47(phox) knockdown . These results suggest that aldosterone inhibits P29474 function through bimodal mechanisms of BH(4) deficiency and PP2A activation . Analysis of neurogenic contractions induced by ML-1035 and other benzamides in the guinea-pig non-stimulated isolated ileum . 4-Amino-5-chloro-substituted benzamides have been shown to increase gastric motility in-vivo and enhance field-stimulated and peristaltic contractions in-vitro . The present experiments examined the contractile response to a series of benzamides in the guinea-pig non-stimulated ileum . Four benzamides elicited contractions in the isolated ileum which were expressed as a percentage of the contraction induced by 1 microM acetylcholine ( % acetylcholine response = 12 +/- 2 , 19 +/- 3 , 26 +/- 2 , 51 +/- 3 , n = 13 , 8 , 17 , and 21 , with EC50 values of 0.85 , 1.8 , 5.7 , and 14.2 microM for cisapride , zacopride , metoclopramide , and ML-1035 ( 4-amino-5-chloro-2-((2-methylsulphinyl)-ethoxy)-N- ( 2-(diethylamino)-ethyl ) -benzamide hydrochloride ) , respectively ) . ML-1035 contractions were completely blocked by atropine and tetrodotoxin , while ganglionic blockade with hexamethonium was ineffective . DB01233 has been reported to sensitize postjunctional muscarinic receptors , however , ML-1035 did not enhance acetylcholine-induced contractions . Tropisetron ( ICS 205-930 , 1 microM ) , caused a parallel rightward shift in the concentration-response curve for both ML-1035 and zacopride ( EC50 = 14.2 +/- 1.3 and 1.8 +/- 0.8 microM in the absence , and 26 +/- 2.7 and 6.9 +/- 2.3 microM in the presence of tropisetron for ML-1035 and zacopride , respectively ) with apparent pKB values of 5.9 and 6.0 for the respective compounds . 5-Hydroxytryptaminergic receptor desensitization by 2-methyl-5-hydroxytryptamine ( 5- Q9H205 ) and 5-methoxytryptamine ( Q13639 ) , attenuated the response to ML-1035. ( ABSTRACT TRUNCATED AT 250 WORDS )

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Use of a cyclo-oxygenase 2 inhibitor for prophylaxis of cystoid macular oedema following cataract surgery : a randomized placebo-controlled trial . BACKGROUND : To assess the efficacy of Celecoxib , a cyclo-oxygenase 2 ( P35354 ) inhibitor , as prophylaxis for cystoid macular oedema after routine cataract surgery . METHODS : A prospective , randomized , double-blind placebo-controlled trial of 69 hospital patients undergoing cataract surgery . Celecoxib 200 mg twice daily or placebo was given immediately after surgery for 14 days . Optical coherence tomography was used to quantify macular thickness before surgery and on day 1 , week 2 and week 6 after surgery . RESULTS : Sixty-nine patients were enrolled , of which 33 received placebo and 36 received active drug . Clinically apparent cystoid macular oedema occurred in four of the treatment group and two of the placebo group ( P = 0.68 ) . No difference in best-corrected visual acuity was seen at 6 weeks ( P = 0.37 ) . Covariate analysis of the results at 2 weeks and 6 weeks showed a macular thickness of 3 % less in the treatment group compared with placebo ( P = 0.050 ) . CONCLUSION : Celecoxib may decrease macular thickening following routine cataract surgery at 2 and 6 weeks after surgery as measured by Stratus O75051 III . No difference in best-corrected visual acuity or clinically apparent cystoid macular oedema was seen . Further investigation of P35354 inhibitors in a larger prospective randomized trial is required . Effects of the inhibition of cyclo-oxygenase 1 or 2 or P09917 on the activation of the hypothalamic-pituitary-adrenal axis induced by interleukin-1beta in the male Rat . The limited entry of interleukin-1beta ( IL-1beta ) into the central nervous system has led to the hypothesis that IL-1beta acts , through IL-1beta receptors located notably on endothelial cells , on the release of prostaglandins which in turn stimulate the hypothalamic-pituitary-adrenal ( Q9Y251 ) axis . We used cyclo-oxygenase-1 ( P23219 ) and cyclo-oxygenase-2 ( P35354 ) and P09917 ( 5- P28300 ) inhibitors , before the injection of IL-1beta , to explore the role of arachidonic acid metabolic pathways on Q9Y251 axis activation . Adult male rats were i.m injected 20 min before i.p injection of IL-1beta , with ( i ) : a P23219 / P35354 inhibitor ( ketoprofen ) ; ( ii ) a P35354 selective inhibitor ( NS 398 ) ; or ( iii ) a 5- P28300 inhibitor ( BW A4C ) . Following this , rats were killed 90 min after i.p . IL-1beta injection and analysis for plasma adrenocorticotropic hormone ( DB01285 ) and corticosterone concentrations and determination of anterior pituitary pro-opio melanocortin ( P01189 ) gene transcription was conducted . Administration of the P23219 / P35354 inhibitor led to a complete blockage of DB01285 and corticosterone secretion and P01189 gene transcription . The P35354 inhibitor led to a complete blockade of DB01285 secretion and P01189 gene transcription but had no effect on corticosterone secretion . The 5- P28300 inhibitor had no significant effect on any parameter . These results demonstrate the crucial role of eicosanoid pathways in mediating the stimulation of the Q9Y251 axis induced by IL-1beta . Moreover , we found a clear dissociation of the effect of the blockage of COXs upon DB01285 and corticosterone secretion , suggesting that IL-1beta may act at the brain as well as at the adrenal cortex to stimulate the secretion of corticosterone . Ocular pharmacokinetics of a single dose of bromfenac sodium ophthalmic solution 0.1 % in human aqueous humor . PURPOSE : The aim of this study was to evaluate the ocular pharmacokinetics of a single dose of bromfenac sodium ophthalmic solution 0.1 % in subjects undergoing routine cataract surgery with intraocular lens implantation . METHODS : An open-label , phase II confirmatory study of 54 subjects undergoing cataract surgery with intraocular lens implantation . A single drop of bromfenac sodium ophthalmic solution 0.1 % was administered at 30 , 60 , 90 , 120 , 180 , or 240 min prior to the initiation of cataract surgery . Samples of aqueous humor were aspirated through a paracentesis and analyzed by using high-performance liquid chromatography . Based upon these data , predicted concentrations of bromfenac in the aqueous humor over 24 h were generated by using computer simulation and compared with the IC(50) for bromfenac as a measure of anti-inflammatory efficacy . RESULTS : Peak aqueous-humor concentrations of bromfenac occurred between 150 and 180 min following ophthalmic dosing , with a mean concentration of 78.7 ng/mL . The level of bromfenac decreased in a log-linear fashion with an elimination-rate constant of 1.4 . DB00963 remained above the IC(50) value of cyclo-oxygenase-2 ( P35354 ) during the evaluated time points and over the 12-h dosing interval , using a computer model of extrapolated time points and assuming first-order elimination . CONCLUSIONS : Pharmacokinetic modeling , based upon samples collected over 240 min after a single dose of bromfenac sodium ophthalmic solution 0.1 % suggests that aqueous-humor concentrations remain at clinically effective levels ( above its IC(50) value for P35354 ) for over 12 h . Based upon this rationale , these results supported clinical trials that demonstrated the efficacy of twice-daily dosing of bromfenac sodium ophthalmic solution 0.1 % to manage patients with postoperative ocular pain and inflammation . P11362 -induced epithelial to mesenchymal transition through MAPK/PLCγ/ P35354 -mediated mechanisms . Tumour invasion and metastasis is the most common cause of death from cancer . For epithelial cells to invade surrounding tissues and metastasise , an epithelial-mesenchymal transition ( EMT ) is required . We have demonstrated that P11362 expression is increased in bladder cancer and that activation of P11362 induces an EMT in urothelial carcinoma ( UC ) cell lines . Here , we created an in vitro P11362 -inducible model of EMT , and used this model to identify regulators of urothelial EMT . P11362 activation promoted EMT over a period of 72 hours . Initially a rapid increase in actin stress fibres occurred , followed by an increase in cell size , altered morphology and increased migration and invasion . By using site-directed mutagenesis and small molecule inhibitors we demonstrated that combined activation of the mitogen activated protein kinase ( MAPK ) and phospholipase C gamma ( PLCγ ) pathways regulated this EMT . Actin stress fibre formation was regulated by PLCγ activation , and was also important for the increase in cell size , migration and altered morphology . MAPK activation regulated migration and P12830 expression , indicating that combined activation of PLCγ and MAPK is required for a full EMT . We used expression microarrays to assess changes in gene expression downstream of these signalling cascades . P35354 was transcriptionally upregulated by P11362 and caused increased intracellular prostaglandin E(2) levels , which promoted migration . In conclusion , we have demonstrated that P11362 activation in UC cells lines promotes EMT via coordinated activation of multiple signalling pathways and by promoting activation of prostaglandin synthesis . Biological differences between in vitro produced bovine embryos and parthenotes . Parthenotes may represent an alternate ethical source of stem cells , once biological differences between parthenotes and embryos can be understood . In this study , we analyzed development , trophectoderm ( TE ) differentiation , apoptosis/necrosis , and ploidy in parthenotes and in vitro produced bovine embryos . Subsequently , using real-time PCR , we analyzed the expression of genes expected to underlie the observed differences at the blastocyst stage . In vitro matured oocytes were either fertilized or activated with ionomycin +6-DMAP and cultured in simple medium . Parthenotes showed enhanced blastocyst development and diploidy and reduced TE cell counts . Apoptotic and necrotic indexes did not vary , but parthenotes evidenced a higher relative proportion of apoptotic cells between inner cell mass and TE . The pluripotence-related Q01860 and the methylation Q9Y6K1 genes were downregulated in parthenotes . Among pregnancy recognition genes , TP-1 was upregulated in parthenotes , while O00264 and PLAC8 did not change . Expression of p66(shc) and Q07812 / P10415 ratio were higher , and p53 lower , in parthenotes . Among metabolism genes , P11166 was downregulated , while P15121 , P35354 , O95479 , and P10599 were upregulated in parthenotes , and P22732 did not differ . Among genes involved in compaction/blastulation , P17302 was downregulated in parthenotes , but no differences were detected within P05023 and CDH1 . Within parthenotes , the expression levels of P11166 , TP-1 , and O95479 , and possibly P15121 , resemble patterns described in female embryos . The pro-apoptotic profile is more pronounced in parthenotes than in embryos , which may differ in their way to channel apoptotic stimuli , through p66(shc) and p53 respectively , and in their mechanisms to control pluripotency and de novo methylation . Chemopreventive efficacy and mechanism of licofelone in a mouse lung tumor model via aspiration . Our previous study comparing inhalation and aspiration to administer agents directly to lung indicated that aspiration route is as effective as inhalation while reducing costs for equipment and chemopreventive agent . This study evaluated the chemopreventive efficacy and mechanism of licofelone , a dual inhibitor of P35354 and P09917 ( 5-Lox ) , via oropharyngeal aspiration against mouse lung adenoma . Eight-week-old female A/J mice were given three doses of benzo[a]pyrene ( B[a]P ; 2 mg/dose , gavage ) to induce lung adenomas . After dysplasia developed , the mice were given licofelone ( 0 , 0.03 , 0.1 , or 0.3 mg/kg ) for 16 weeks , and tumor incidence and multiplicity in lung were measured . In addition , the expression of a series of biomarkers in lung cancer progression was evaluated at 2 and 16 weeks . DB04725 showed dose-related inhibition of B[a]P-induced tumor incidence and multiplicity at 0.03 and 0.1 mg/kg following 16-week treatment . DB04725 also showed dose-dependent inhibition of P35354 ( 25 % -41 % ) and 5-Lox ( 35 % -61 % ) at 2 and 16 weeks and proliferating cell nuclear antigen ( P12004 ; 41 % -61 % ) at 16 weeks . A dose-dependent increase in apoptosis ( 1.5- to 2.4-fold ) was also observed in licofelone groups . A marginal inhibition of survivin was observed at one dose . In conclusion , this study showed that licofelone via aspiration showed chemopreventive efficacy against mouse lung adenoma with good correlation to early and late biomarkers of lung cancer progression . This is the first study to show that the aspiration route can be an excellent inexpensive alternative to inhalation for direct delivery of drugs to rodent lungs for efficacy testing of potential chemopreventive agents . [ DB00054 ( ReoPro ) in the treatment of acute coronary syndromes ] . Platelet activation plays a major role in the pathophysiology of acute coronary syndromes ( ACS ) . Inhibition of platelet function is the basic pharmacological treatment of ACS . P08514 /IIIa inhibitors , a new class of potent antiplatelet agents , have been used in the treatment of ACS and in the prevention of complications after percutaneous coronary interventions ( P05154 ) . Several large clinical trials have demonstrated the effectiveness of this class of agents . The first of these agents to show beneficial effects after coronary interventions was the mouse/human chimeric Fab fragment antibody c7E3 abciximab ( ReoPro ) . The purpose of this article is to describe the pharmacology of abciximab and to review the results of the clinical trials carried out with the drug in patients with ACS , treated either with or without acute/elective P05154 . P20809 induces osteoblast differentiation and acts synergistically with bone morphogenetic protein-2 in C3H10T1/2 cells . P20809 ( IL-11 ) is a pleiotropic cytokine that supports various types of hematopoietic cell growth and is involved in bone resorption . We report here the involvement of recombinant human IL-11 ( rHuIL-11 ) in osteoblast differentiation in mouse mesenchymal progenitor cells , C3H10T1/2. rHuIL-11 alone increased alkaline phosphatase ( ALP ) activity and upregulated expression levels of osteocalcin ( OC ) , bone sialo protein ( BSP ) , and parathyroid hormone receptor ( Q03431 ) mRNA. rHuIL-11 had no effect on expression of type II collagen , peroxisome proliferator-activated receptor-gamma2 ( Q07869 -gamma2 ) , adipocyte fatty acid-binding protein P2 ( aP2 ) , and myogenic MyoD protein ( MyoD ) . Recombinant human bone morphogenetic protein ( rHuBMP ) -2 increased ALP activity and mRNA expression of these genes except for MyoD . The expression patterns of ALP activity and osteoblast-specific or chondrocyte-specific genes suggest that rHuIL-11 may be involved in early differentiation of osteoblasts at a step earlier than that which is affected by rHuBMP-2 . In support of this hypothesis , combined treatment with rHuIL-11 and rHuBMP-2 synergistically increased ALP activity and mRNA expression of OC and type II collagen , rHuIL-11 also abrogated the increased levels of Q07869 -gamma2 , aP2 mRNA caused by rHuBMP-2 . Our results suggest that rHuIL-11 alone and in combination with rHuBMP-2 can induce osteoblastic differentiation of progenitor cells and plays an important role in osteogenesis . Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared . P05305 induced proinflammatory markers in the myocardium and leukocytes of guinea-pigs : role of glycoprotein IIB/IIIA receptors . AIM : To assess whether endothelin-1 ( ET-1 ) induces the in vivo expression of inflammatory-related proteins , namely cyclooxygenase-2 ( P35354 ) and tissue factor , in the myocardium and circulating leukocytes of guinea-pigs . The involvement of platelets was also analyzed . METHODS : ET-1 ( 0.013 microg/min ) was infused to male guinea-pigs for 45 min in the presence and absence of tirofiban , a nonpeptidic blocker of the glycoprotein IIb/IIIa receptor ( P08514 /IIIa ) . P13726 and P35354 expression were determined by Western blot . RESULTS : No changes in mean arterial pressure and heart rate were detected . ET-1-infused guinea-pigs showed a marked increase in the number of platelets expressing activated P08514 /IIIa receptors ( 0.8+/-0.03 % vs. 6.5+/-0.2 % ; P < 0.05 ) . Tirofiban ( 10 microg/Kg bw/min ) blunted ex vivo platelet aggregation in response to ADP , although only partially reduced P35354 and tissue factor expression in both the myocardium and leukocytes of ET-1-infused guinea-pigs . The myocardium of platelet-depleted guinea-pigs also showed a reduced P35354 expression after ET-1 infusion ( 57+/-3 % reduction ; P < 0.05 ) . In vitro studies demonstrated that platelets ( 10(7) and 10(9) platelets/well ) enhanced ET-1 ( 10(-7) mol/l ) -induced P35354 expression in heart slices . CONCLUSION : ET-1 stimulated in vivo the expression of the pro-inflammatory proteins P35354 and tissue factor in the myocardium and in leukocytes by a mechanism P08514 /IIIa platelet receptors . Effect of milk hydrolysates on inflammation markers and drug-induced transcriptional alterations in cell-based models . Nonsteroidal anti-inflammatory drugs ( NSAID ) are associated with gastrointestinal inflammation and subsequent damage to the intestinal tissue . Earlier studies in our laboratory have found that specific casein hydrolysates ( CH ) might be useful in the treatment of gastrointestinal wounds . The underlying mechanisms that support inflammation and wound healing are not completely understood , but transcriptional alterations may be used as markers for inflammation and wound healing . The bioactivity of 3 CH prepared by treatment of commercial casein with pepsin ( 60 min ) followed by corolase ( 0 , 10 , or 60 min ) were investigated in intestinal epithelial cells treated with the NSAID indomethacin . The bioactivity was evaluated as transcriptional alterations of transforming growth factor-β1 ( TGF-β1 ) , cyclooxygenase-2 ( P35354 ) , peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) and nuclear factor κB ( NFκB ) by real-time PCR . Furthermore , the effect of CH on lipopolysaccharide-induced inflammation was evaluated in macrophages by measuring PG E(2) levels . Casein hydrolysates treated with corolase for 10 or 60 min after pepsin treatment downregulated transcription of TGF-β1 and NFκB ( P < 0.05 ) compared with the hydrolysate treated with pepsin only . Hydrolysate prepared by corolase treatment for 60 min after pepsin hydrolysis downregulated transcription of P35354 ( P < 0.05 ) compared with hydrolysate treated with corolase for only 10 min whereas transcription of Q07869 -γ was not affected ( P > 0.05 ) . Additionally , the hydrolysate prepared by pepsin treatment only ( 0 min corolase ) had a pro-inflammatory effect on macrophages via PG E(2) stimulation ( P < 0.05 ) . In conclusion , CH produced by a combination of pepsin and corolase treatments downregulated the transcription levels of TGF-β1 , P35354 , and NFκB . DB06698 ameliorates olanzapine-induced weight gain through modulation of histaminergic , P01303 and AMPK pathways . Olanzapine is widely used to treat schizophrenia and other disorders , but causes adverse obesity and other metabolic side-effects . Both animal and clinical studies have shown that co-treatment with betahistine ( a histaminergic H1 receptor agonist and H3 receptor antagonist ) is effective for ameliorating olanzapine-induced weight gain/obesity . To reveal the mechanisms underlying these effects , this study investigated the effects of co-treatment of olanzapine and betahistine ( O+B ) on expressions of histaminergic H1 receptor ( P35367 ) , AMP-activated protein kinase ( AMPK ) , neuropeptide Y ( P01303 ) , and proopiomelanocortin ( P01189 ) in the hypothalamus associated with reducing olanzapine-induced weight gain . Olanzapine significantly upregulated the mRNA and protein expressions of P35367 , while O+B co-treatment significantly downregulated the P35367 levels , compared to the olanzapine-only treatment group . The P01303 mRNA expression was significantly enhanced by olanzapine , but it was significantly reversed by O+B co-treatment . The hypothalamic P35367 expression was positively correlated with total food intake , and P01303 expression . Olanzapine also increased AMPKα activation measured by the AMPKα phosphorylation ( pAMPKα ) /AMPKα ratio compared with controls , whereas O+B co-treatment decreased the pAMPKα/AMPKα ratio , compared with olanzapine only treatment . The pAMPKα/AMPKα ratio was positively correlated with total food intake and P35367 expression . Although olanzapine administration decreased the P01189 mRNA level , this level was not affected by O+B co-treatment . Therefore , these results suggested that co-treatment with betahistine may reverse olanzapine-induced body weight gain via the P35367 - P01303 and P35367 -pAMPKα pathways . Inhibition of tumor cell growth , invasion , and metastasis by EXEL-2880 ( DB05030 , GSK1363089 ) , a novel inhibitor of P14210 and P15692 receptor tyrosine kinases . The DB00134 receptor tyrosine kinase and its ligand , hepatocyte growth factor ( P14210 ) , are overexpressed and/or activated in a wide variety of human malignancies . Vascular endothelial growth factor ( P15692 ) receptors are expressed on the surface of vascular endothelial cells and cooperate with DB00134 to induce tumor invasion and vascularization . EXEL-2880 ( DB05030 , GSK1363089 ) is a small-molecule kinase inhibitor that targets members of the P14210 and P15692 receptor tyrosine kinase families , with additional inhibitory activity toward P10721 , Flt-3 , platelet-derived growth factor receptor beta , and Tie-2 . Binding of EXEL-2880 to DB00134 and P15692 receptor 2 ( P35968 ) is characterized by a very slow off-rate , consistent with X-ray crystallographic data showing that the inhibitor is deeply bound in the DB00134 kinase active site cleft . EXEL-2880 inhibits cellular P14210 -induced DB00134 phosphorylation and P15692 -induced extracellular signal-regulated kinase phosphorylation and prevents both P14210 -induced responses of tumor cells and P14210 / P15692 -induced responses of endothelial cells . In addition , EXEL-2880 prevents anchorage-independent proliferation of tumor cells under both normoxic and hypoxic conditions . In vivo , these effects produce significant dose-dependent inhibition of tumor burden in an experimental model of lung metastasis . Collectively , these data indicate that EXEL-2880 may prevent tumor growth through a direct effect on tumor cell proliferation and by inhibition of invasion and angiogenesis mediated by P14210 and P15692 receptors . Synthesis and biological evaluation of novel oxindole-based RTK inhibitors as anti-cancer agents . Given that receptor tyrosine kinases ( RTKs ) have emerged as key regulators of all aspects of cancer development , including proliferation , invasion , angiogenesis and metastasis , the RTK family represents an important therapeutic target for anti-cancer drug development . Oxindole structure has been used in RTK inhibitors such as DB02058 and intedanib . In this study , two series of new heterocyclic compounds containing oxindole scaffold have been designed and synthesized , and their inhibitory activity against the proliferation of nine cancer cell lines has been evaluated . Among them , compounds 9a and 9b displayed the strongest anti-proliferative activity with the IC50s below 10μM . Flow cytometric analysis showed that the compounds 9a and 9b dose-dependently arrested the cell cycle at G0/ P55008 phase . Although the leading compounds DB02058 and intedanib targets P11362 , the kinase activity test revealed that these compounds only showed slight inhibitory activity on P11362 kinase . Further enzymatic test aided by molecular docking simulation in the DB00171 -binding site demonstrated that 9a and 9b are potent inhibitors of c-Kit kinase . These compounds are worthy of further evaluation as anticancer agents . 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 . P28562 regulates bone mass , osteoblast gene expression , and responsiveness to parathyroid hormone . Parathyroid hormone ( PTH ) signaling via PTH 1 receptor ( Q03431 ) involves mitogen-activated protein kinase ( MAPK ) pathways . MAPK phosphatase 1 ( P28562 ) dephosphorylates and inactivates MAPKs in osteoblasts , the bone-forming cells . We previously showed that Q03431 activation in differentiated osteoblasts upregulates P28562 and downregulates pERK1/2-MAPK and cyclin D1 . In this study , we evaluated the skeletal phenotype of Mkp1 knockout ( KO ) mice and the effects of PTH in vivo and in vitro . Microcomputed tomography analysis of proximal tibiae and distal femora from 12-week-old Mkp1 KO female mice revealed osteopenic phenotype with significant reduction ( 8-46 % ) in bone parameters compared with wild-type ( WT ) controls . Histomorphometric analysis showed decreased trabecular bone area in KO females . Levels of serum osteocalcin ( OCN ) were lower and serum tartrate-resistant acid phosphatase 5b ( TRAP5b ) was higher in KO animals . Treatment of neonatal mice with DB05829 ( 1-34 ) for 3 weeks showed attenuated anabolic responses in the distal femora of KO mice compared with WT mice . Primary osteoblasts derived from KO mice displayed delayed differentiation determined by alkaline phosphatase activity , and reduced expressions of Ocn and Runx2 genes associated with osteoblast maturation and function . Cells from KO females exhibited attenuated PTH response in mineralized nodule formation in vitro . Remarkably , this observation was correlated with decreased PTH response of matrix Gla protein expression . Expressions of pERK1/2 and cyclin D1 were inhibited dramatically by PTH in differentiated osteoblasts from WT mice but much less in osteoblasts from Mkp1 KO mice . In conclusion , P28562 is important for bone homeostasis , osteoblast differentiation and skeletal responsiveness to PTH . Vitamin D analogues . The plethora of actions attributed to 1,25(OH)2D3 throughout the body have suggested potential therapeutic applications for the treatment of hyperproliferative diseases , immune dysfunction , endocrine disorders , and metabolic bone disease . However , the potent calcemic activity of the natural vitamin D hormone has precluded its use in most cases . New vitamin D analogues are under development that display greater specificity , in most cases , by retaining the therapeutic properties of 1,25(OH)2D3 , but with lower calcemic activity . Two analogues have been approved for use in patients : calcipotriol ( DB02300 from Leo Pharmaceuticals , Copenhagen , Denmark ) for the treatment of psoriasis ; and 19-nor-1,25(OH)2D2 ( DB00910 from Abbott Laboratories , Abbott Park , IL ) for secondary hyperparathyroidism . Many others analogues are currently being tested in preclinical and clinical trials for the treatment of various types of cancer and osteoporosis , and for immunosuppression . The selectivity of the analogues can be attributed to the combined interactions with four proteins : the vitamin D receptor ( P11473 ) , the serum vitamin D binding protein ( DBP ) , the vitamin D-24-hydroxylase and to a newly described membrane receptor . Low DBP affinity has been shown to be responsible for the reduced calcemic actions of calcipotriol and 22-oxacalcitriol ( O75051 ) , which is being tested for secondary hyperparathyroidism . However , the low calcemic activity of other analogues , including 19-nor-1,25(OH)2D2 , involves other , as yet undefined , mechanisms . Understanding of the molecular basis for the selectivity of the vitamin D analogues will allow the design of more effective and safer vitamin D compounds for the treatment of a wide range of clinical disorders . P10415 /BCL-X(L) inhibition induces apoptosis , disrupts cellular calcium homeostasis , and prevents platelet activation . Apoptosis in megakaryocytes results in the formation of platelets . The role of apoptotic pathways in platelet turnover and in the apoptotic-like changes seen after platelet activation is poorly understood . DB05764 ( Navitoclax ) , a specific inhibitor of antiapoptotic P10415 proteins , which is currently being evaluated in clinical trials for the treatment of leukemia and other malignancies , induces a dose-limiting thrombocytopenia . In this study , the relationship between P10415 /BCL-X(L) inhibition , apoptosis , and platelet activation was investigated . Exposure to DB05764 induced apoptosis but repressed platelet activation by physiologic agonists . Notably , DB05764 induced an immediate calcium response in platelets and the depletion of intracellular calcium stores , indicating that on P10415 /BCL-X(L) inhibition platelet activation is abrogated because of a diminished calcium signaling . By comparing the effects of DB05764 and its analog ABT-737 on platelets and leukemia cells from the same donor , we show , for the first time , that these P10415 /BCL-X(L) inhibitors do not offer any selective toxicity but induce apoptosis at similar concentrations in leukemia cells and platelets . However , reticulated platelets are less sensitive to apoptosis , supporting the hypothesis that treatment with DB05764 induces a selective loss of older platelets and providing an explanation for the transient thrombocytopenia observed on DB05764 treatment .

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Thrombin receptors and their antagonists : an update on the patent literature . IMPORTANCE OF THE FIELD : Thrombin plays a central role in cardiovascular inflammation . Most of the cellular responses to thrombin are mediated by cell surface protease-activated receptors ( PARs ) . Several preclinical studies indicate that PARs are potential targets for treating cardiovascular diseases such as thrombosis , atherosclerosis and restenosis . Among PARs , P25116 has emerged as an important therapeutic target . AREAS COVERED IN THIS REVIEW : This review covers recent advances in the development of thrombin receptors antagonists . It is focused on the search for P25116 antagonists as this is at the moment the most promising and attractive target . However , some early promising studies on PAR-3 and -4 antagonists are also reported . WHAT THE READER WILL GAIN : The review has been written in order to give to the reader hints and references that cover , in our opinion , the most interesting and/or promising approaches in this research field . TAKE HOME MESSAGE : Research on P25116 antagonists has finally led to good clinical candidates such as DB05692 ( Schering-Plough ) and E-5555 ( Eisai Co. ) . Clinical trials clearly demonstrate that development of PAR1 antagonists is not only possible but most likely will lead to development of antiplatelet drugs as well as of drugs useful for the treatment of inflammatory , proliferative and neurodegenerative diseases . DB00877 inhibits poly(ADP-ribosyl)ation in intact cells . DB00877 is an immunosuppressive drug , which inhibits the mammalian target of rapamycin ( P42345 ) kinase activity inducing changes in cell proliferation . Synthesis of poly(ADP-ribose) ( PAR ) is an immediate cellular response to genotoxic stress catalyzed mostly by poly(ADP-ribose) polymerase 1 ( P09874 ) , which is also controlled by signaling pathways . Therefore , we investigated whether rapamycin affects PAR production . Strikingly , rapamycin inhibited PAR synthesis in living fibroblasts in a dose-dependent manner as monitored by immunofluorescence . P09874 activity was then assayed in vitro , revealing that down-regulation of cellular PAR production by rapamycin was apparently not due to competitive P09874 inhibition . Further studies showed that rapamycin did not influence the cellular NAD pool and the activation of P09874 in extracts of pretreated fibroblasts . Collectively , our data suggest that inhibition of cellular PAR synthesis by rapamycin is mediated by formation of a detergent-sensitive complex in living cells , and that rapamycin may have a potential as therapeutic PARP inhibitor . Thrombin induces osteoprotegerin synthesis via phosphatidylinositol 3'-kinase/mammalian target of rapamycin pathway in human periodontal ligament cells . BACKGROUND AND OBJECTIVE : Thrombin influences the biological behavior of periodontal ligament cells and plays multiple roles in the early stages of bone healing . O00300 ( O00300 ) is one of the key molecules that regulate bone homeostasis and prevent osteoclastogenesis . The purpose of this study was to evaluate the biological effects of thrombin on O00300 synthesis in human periodontal ligament ( Q96IR7 ) cells in vitro . MATERIAL AND METHODS : Cells were treated with various concentrations ( 0.001 , 0.01 and 0.1 U/mL ) of thrombin . The mRNA expression and protein synthesis of O00300 , as well as of receptor activator of nuclear factor kappaB ligand ( O14788 ) , were determined by reverse transcriptase-polymerase chain reaction ( RT-PCR ) and Western blot analysis , respectively . The influence of thrombin on O00300 synthesis and its signaling pathway were investigated using inhibitors . RESULTS : Thrombin profoundly induces protein synthesis of O00300 at 0.1 U/mL . The inductive effect was inhibited by cycloheximide , but not by indomethacin . The phosphatidylinositol 3'-kinase ( PI3K ) inhibitor , LY294002 , and the mammalian target of rapamycin ( P42345 ) inhibitor , rapamycin , exerted an inhibitory effect on the thrombin-induced O00300 synthesis . In addition , the effect was inhibited by protease-activated receptor ( PAR ) -1 antagonist . Activation of phospho-Akt ( p-Akt ) was observed and the effect was abolished by LY294002 . CONCLUSION : Thrombin induces O00300 synthesis in Q96IR7 cells post-transcriptionally , possibly through P25116 . The regulation was through the PI3K/Akt and P42345 pathway . This finding suggests that thrombin may play a significant role in alveolar bone repair and homeostasis of periodontal tissue , partly through the O00300 / O14788 system . DB08836 treatment ameliorates autoimmune myocarditis associated with enhanced cardiomyocyte thioredoxin expression . P10599 ( TRX ) is a redox regulatory protein that protects cells from various stresses . P12821 ( P12821 ) inhibitor was reported to enhance endogenous antioxidant enzyme activities . This study was carried out to investigate whether temocapril , a novel non-sulfhydryl containing P12821 inhibitor , reduces the severity of myocarditis via redox regulation mechanisms involving TRX . Western blot showed that temocapril enhanced cytosolic redox regulatory protein TRX expression , but neither mitochondrial TRX2 nor antioxidant enzymes , such as copper-zinc superoxide dismutase ( Cu/Zn-SOD ) or manganese superoxide dismutase ( Mn-SOD ) expression , was increased by the preconditioning treatment . In rats with experimental autoimmune myocarditis ( EAM ) , the protein carbonyl content , a marker of cellular protein oxidation , was increased accompanied with enhanced TRX expression . An immunohistochemical study showed that TRX stain was enhanced in infiltrating inflammatory cells and in damaged myocytes . The severity of the myocarditis and the protein carbonyl contents were less increased in temocapril treatment ( 10 mg/kg/day , orally ) from day 1 to day 21 in which TRX was up regulated when the inflammation started , but not in temocapril treatment from day 15-21 in which TRX was not up-regulated when the inflammation started . The results suggest that TRX and the redox state modified by TRX may play a crucial role in the pathophysiology of EAM . DB08836 ameliorates myocarditis associated with inducing TRX increase in a preconditioning manner , although the mechanism of TRX induction by temocapril remains to be elucidated . Design and synthesis of 3,5-disubstituted-1,2,4-oxadiazoles as potent inhibitors of phosphodiesterase4b2 . A series of 3,5-disubstituted-1,2,4-oxadiazoles has been prepared and evaluated for phosphodiesterase inhibition ( PDE4B2 ) . Among the prepared 3,5-disubstituted-1,2,4-oxadiazoles , compound 9a is the most potent inhibitor ( PDE4B2 IC(50) = 5.28 μm ) . Structure-activity relationship studies of 3,5-disubstituted-1,2,4-oxadiazoles revealed that substituents 3-cyclopentyloxy-4-methoxyphenyl group at 3-position and cyclic ring bearing heteroatoms at 5-position are important for activity . Molecular modeling study of the 3,5-disubstituted-1,2,4-oxadiazoles with Q07343 has shown similar interactions of 3-cyclopentyloxy-4-methoxyphenyl group ; however , heteroatom ring is slightly deviating when compared to DB01791 . 3-(3-Cyclopentyloxy-4-methoxyphenyl)-5-(piperidin-4-yl)-1,2,4-oxadiazole ( 9a ) exhibited good analgesic and antiinflammatory activities in formalin-induced pain in mice and carrageenan-induced paw edema model in rat . Inducible raptor and rictor knockout mouse embryonic fibroblasts . The mammalian Target of DB00877 ( P42345 ) kinase functions within two structurally and functionally distinct multiprotein complexes termed P42345 complex 1 ( mTORC1 ) and mTORC2 . The immunosuppressant and anticancer drug rapamycin is commonly used in basic research as a tool to study P42345 signaling . However , rapamycin inhibits only , and only incompletely , mTORC1 , and no mTORC2-specific inhibitor is available . Hence , a full understanding of P42345 signaling in vivo , including the function of both complexes , requires genetic inhibition in addition to pharmacological inhibition . Taking advantage of the Cre/LoxP system , we generated inducible knockout mouse embryonic fibroblasts ( MEFs ) deficient for either the mTORC1-specific component raptor ( iRapKO ) or the mTORC2-specific component rictor ( iRicKO ) . Inducibility of the knockout was important because P42345 complex components are essential . Induction of either raptor or rictor knockout eliminated raptor or rictor expression , respectively , and impaired the corresponding P42345 signaling branch . The described knockout MEFs are a valuable tool to study the full function of the two P42345 complexes individually . DB00877 unbalances the polarization of human macrophages to M1 . Plasticity is a hallmark of macrophages , and in response to environmental signals these cells undergo different forms of polarized activation , the extremes of which are called classic ( M1 ) and alternative ( M2 ) . DB00877 ( Q96PN7 ) is crucial for survival and functions of myeloid phagocytes , but its effects on macrophage polarization are not yet studied . To address this issue , human macrophages obtained from six normal blood donors were polarized to M1 or M2 in vitro by lipopolysaccharide plus interferon-γ or interleukin-4 ( P05112 ) , respectively . The presence of Q96PN7 ( 10 ng/ml ) induced macrophage apoptosis in M2 but not in M1 . Beyond the impact on survival in M2 , Q96PN7 reduced P61073 , CD206 and Q9NNX6 expression and stem cell growth factor-β , P55774 and Q99616 release . In contrast , in M1 Q96PN7 increased P42081 and P32248 expression and P05231 , tumour necrosis factor-α and IL-1β release but reduced CD206 and Q9NNX6 expression and P22301 , vascular endothelial growth factor and P55774 release . In view of the in vitro data , we examined the in vivo effect of Q96PN7 monotherapy ( 0·1 mg/kg/day ) in 12 patients who were treated for at least 1 month before islet transplant . Cytokine release by O00206 -stimulated peripheral blood mononuclear cells showed a clear shift to an M1-like profile . Moreover , macrophage polarization 21 days after treatment showed a significant quantitative shift to M1 . These results suggest a role of mammalian target of rapamycin ( P42345 ) into the molecular mechanisms of macrophage polarization and propose new therapeutic strategies for human M2-related diseases through P42345 inhibitor treatment . P38936 (Waf1) is required for cellular senescence but not for cell cycle arrest induced by the HDAC inhibitor sodium butyrate . Cell senescence is characterized by senescent morphology and permanent loss of proliferative potential . HDAC inhibitors ( HDACI ) induce senescence and/or apoptosis in many types of tumor cells . Here , we studied the role of cyclin-kinase inhibitor P38936 (waf1) ( Cdkn1n gene ) in cell cycle arrest , senescence markers ( cell hypertrophy , SA-βGal staining and accumulation of γ P16104 foci ) in P38936 (Waf1+/+) versus P38936 (Waf1-/-) mouse embryonic fibroblast cells transformed with E1A and cHa-Ras oncogenes ( mERas ) . While short treatment with the HDACI sodium butyrate ( NaB ) induced a reversible G(1) cell cycle arrest in both parental and P38936 (Waf1-/-) cells , long-term treatment led to dramatic changes in P38936 (Waf1+/+) cells only : cell cycle arrest became irreversible and cells become hypertrophic , SA-βGal-positive and accumulated γ P16104 foci associated with mTORC1 activation . The P38936 (Waf1+/+) cells lost their ability to migrate into the wound and through a porous membrane . Suppression of migration was accompanied by accumulation of vinculin-staining focal adhesions and Ser3-phosphorylation of cofilin , incapable for F-actin depolymerization . In contrast , the knockout of the P38936 (Waf1) abolished most of the features of NaB-induced senescence , including irreversibility of cell cycle arrest , hypertrophy , additional focal adhesions and block of migration , γ P16104 foci accumulation and SA-βGal staining . DB00877 , a specific inhibitor of mTORC1 kinase , decreased cellular hypertrophy , canceled coffilin phosphorylation and partially restored cell migration in P38936 (Waf1+/+) cells . Taken together , our data indicate a new role of P38936 (Waf1) in cell senescence , which may be connected not only with execution of cell cycle arrest , but also with the development of P42345 -dependent markers of cellular senescence . Korean Red DB01404 Extract Enhances the Anticancer Effects of Imatinib Mesylate Through Abrogation p38 and P42229 Activation in KBM-5 Cells . Although imatinib mesylate ( IM ) in the treatment of chronic myelogenous leukemia ( CML ) remains the best example of successful targeted therapy , majority of patients with CML suffer its toxicity profile and develop chemoresistance to existing therapeutic agents . Thus , there is a need to develop novel alternative therapies for the treatment of CML . Here , we investigated whether Korean red ginseng extract ( KRGE ) could suppress the proliferation and induce chemosensitization in human CML cells . Also , we used a human phospho-antibody array containing 46 antibodies against signaling molecules to examine a subset of phosphorylation events after treatment . Korean red ginseng extract broadly suppressed the proliferation of five different cell lines , but KRGE was found to be the most potent inducer of apoptosis against KBM-5 cells . It also abrogated the expression of Bcl-2 ( B-cell lymphoma 2 ) , Bcl-xL ( B-cell lymphoma-extra large ) , survivin , inhibitors of apoptosis protein 1/2 , P35354 ( P35354 ) , cyclin D1 , matrix metalloproteinase-9 , and P15692 ( Vascular endothelial growth factor ) , as well as upregulated the expression of pro-apoptotic gene products . Interestingly , KRGE also enhanced the cytotoxic and apoptotic effect of IM in KBM-5 cells . The combination treatment of KRGE and IM caused pronounced suppression of p38 and signal transducer and activator of transcription 5 phosphorylation and induced phosphorylation of p53 compared with the individual treatment . Our results demonstrate that KRGE can enhance the anticancer activity of IM and may have a substantial potential in the treatment of CML . Preclinical in vivo evaluation of rapamycin in human malignant peripheral nerve sheath explant xenograft . Neurofibromatosis type 1 ( P21359 ) patients are prone to the development of malignant tumors , the most common being Malignant Peripheral Nerve Sheath Tumor ( MPNST ) . P21359 -MPNST patients have an overall poor survival due to systemic metastasis . Currently , the management of MPNSTs includes surgery and radiation ; however , conventional chemotherapy is not very effective , underscoring the need for effective biologically-targeted therapies . Recently , the P21359 gene product , neurofibromin , was shown to negatively regulate the phosphoinositide-3-kinase ( PI3K ) /Protein Kinase-B ( Akt ) /mammalian Target Of DB00877 ( P42345 ) pathway , with loss of neurofibromin expression in established human MPNST cell lines associated with high levels of P42345 activity . We developed and characterized a human P21359 -MPNST explant grown subcutaneously in NOD-SCID mice , to evaluate the effect of the P42345 inhibitor rapamycin . We demonstrate that rapamycin significantly inhibited human P21359 -MPNST P42345 pathway activation and explant growth in vivo at doses as low as 1.0 mg/kg/day , without systemic toxicities . While rapamycin was effective at reducing P21359 -MPNST proliferation and angiogenesis , with decreased CyclinD1 and P15692 respectively , there was no increase in tumor apoptosis . DB00877 effectively decreased activation of S6 downstream of P42345 , but there was accompanied increased Akt activation . This study demonstrates the therapeutic potential and limitations of rapamycin in P21359 -associated , and likely sporadic , MPNSTs . Inactivation of caspase-1 in rodent brain : a novel anticonvulsive strategy . PURPOSE : Cytokines and related inflammatory mediators are rapidly synthesized in the brain during seizures . We previously found that intracerebral administration of interleukin-1 ( IL-1 ) -beta has proconvulsant effects , whereas its endogenous receptor antagonist ( IL-1Ra ) mediates potent anticonvulsant actions in various models of limbic seizures . In this study , we investigated whether seizures can be effectively inhibited by blocking the brain production of IL-1beta , by using selective inhibitors of interleukin-converting enzyme ( ICE/caspase-1 ) or through caspase-1 gene deletion . METHODS : P29466 was selectively blocked by using pralnacasan or DB05507 . IL-1beta release was induced in mouse organotypic hippocampal slice cultures by proinflammatory stimuli [ lipopolysaccharide ( LPS ) + adenosine triphosphate ( DB00171 ) ] and measured with enzyme-linked immunosorbent assay ( ELISA ) . IL-1beta production during seizures was measured in the rat hippocampus by Western blot . Seizures were induced in freely moving mice and rats by intrahippocampal injection of kainic acid and recorded by EEG analysis . RESULTS : P29466 inhibition reduced the release of IL-1beta in organotypic slices exposed to LPS+ DB00171 . Administration of pralnacasan ( intracerebroventricular , 50 microg ) or DB05507 ( intraperitoneal , 25-200 mg/kg ) to rats blocked seizure-induced production of IL-1beta in the hippocampus , and resulted in a twofold delay in seizure onset and 50 % reduction in seizure duration . Mice with caspase-1 gene deletion showed a 70 % reduction in seizures and an approximate fourfold delay in their onset . CONCLUSIONS : Inhibition of caspase-1 represents an effective and novel anticonvulsive strategy , which acts by selectively reducing the brain availability of IL-1beta . [ Neuroprotective effect of epigallocatechin gallate on oxidative-stress-injured retinal cells ] . OBJECTIVE : To investigate the neuroprotective effect of epigallocatechin gallate(EGCG) , the main extract from green tea , on the oxidative-stress-injured retinal ganglion cells . METHODS : Rat retinal ganglion cells ( RGC-5 ) were cultured into 3 groups ( normal control ; H2O2 ; H2O2 + EGCG or Trolox or DB02690 ) . In-situ TUNEL was used to detect the apoptosis of the RGC-5 cells . Dihydroethidium ( DHE ) assay was used to observe the intracellular ROS generation . The activation of nuclear enzyme , P09874 was quantitatively detected by Western blot and the cell viability was measured by MT method . RESULTS : Hydrogen peroxide reduced RGC-5 cell viability in a time-concentration-dependent manner . The treatment of 500 micromol/L H2O2 for 24 hours reduced RGC-5 cell viability by about 50 % of control . Hydrogen peraoxide caused apoptosis of the RGC-5 cell , obviously increased intracellular ROS generation and up-regulated the P09874 expression . The pretreatment with EGCG was able to markedly reduce the number of apoptotic cells , attenuate intracellular ROS generation . Furthermore , MTT assay showed that the pretreatment with EGCG ( 50 micromol/L ) increased the most cell viability to 87 % of control , but pretreatment with Trolox ( 100 micromol/L ) and DB02690 ( 100 micromol/L , a P09874 inhibitor ) recovered the most cell viability to 62 % and 71 % of control respectively . CONCLUSION : EGCG is able to effectively protect retinal ganglion cell against oxidative-stressed injury and can be used as a very potential neuroprotective drug . The anti-interleukin-6 antibody siltuximab down-regulates genes implicated in tumorigenesis in prostate cancer patients from a phase I study . BACKGROUND : P05231 ( P05231 ) is associated with prostate cancer morbidity . In several experimental models , P05231 has been reported to have anti-apoptotic and pro-angiogenic effects . DB09036 ( CNTO 328 ) is a monoclonal anti- P05231 antibody which has been successfully applied in several models representing prostate cancer . This study was designed to assess preliminary safety of siltuximab in patients with early prostate cancer . PATIENTS AND METHODS : Twenty patients scheduled to undergo radical prostatectomy received either no drug or siltuximab ( 6 mg/kg , five patients per group with administration once , two times , and three times prior to surgery ) . Blood samples were collected for pharmacokinetic and pharmacodynamic analyses . Expression of elements of P05231 signaling pathways was analyzed in tumor tissue by immunohistochemistry . Gene analysis in tumor specimens was performed with the DASL array . RESULTS : No adverse events related to siltuximab were observed . Patients treated with siltuximab presented with higher levels of proliferation and apoptosis markers . Following a single dose , serum concentrations of siltuximab declined in a biexponential manner . This study revealed a decrease in phosphorylation of Stat3 and Q8TCB0 / Q8NFH3 mitogen-activated protein kinases . In addition , gene expression analyses indicate down-regulation of genes immediately downstream of the P05231 signaling pathway and key enzymes of the androgen signaling pathway . CONCLUSIONS : Preliminary safety of siltuximab is favorable . Future studies in which siltuximab could be combined with androgen-deprivation therapy and experimental therapies in advanced prostate cancer are justified . Distinct functional significance of Akt and P42345 constitutive activation in mantle cell lymphoma . Functional characterization of signaling pathways that critically control mantle cell lymphoma ( Q8WXI8 ) cell growth and survival is relevant to designing new therapies for this lymphoma . We herein demonstrate that the constitutive activation of Akt correlates with the expression of the phosphorylated , inactive form of P60484 . Phosphatidyl-inositol-3 kinase ( P19957 -K ) /Akt or mammalian target of rapamycin ( P42345 ) inhibition decreased the growth of both primary Q8WXI8 cultures and established cell lines and antagonizes the growth-promoting activity of P25942 triggering and P05112 . These effects are mediated by nuclear accumulation of the p27(Kip1) inhibitor induced by down-regulation of the P29466 (Skp2) and Cks1 proteins , which target p27(Kip1) for degradation . Moreover , Akt inhibition down-regulated cyclin D1 by promoting its proteasome-dependent degradation driven by GSK-3 . Intriguingly , P42345 inhibition affected cyclin D1 proteolysis only in Q8WXI8 cells in which GSK-3 is under the direct control of P42345 , suggesting that different Q8WXI8 subsets could be differently responsive to P42345 inhibition . Finally , P19957 -K/Akt inhibitors , but not rapamycin , induced variable levels of caspase-dependent apoptosis and reduced telomerase activity . These results indicate that Akt and P42345 activation have distinct functional relevance in Q8WXI8 and suggest that targeting Akt may result in more effective therapeutic effects compared with P42345 inhibition . 1H-1,2,4-triazol-3-yl-anilines : novel potent inhibitors of vascular endothelial growth factor receptors 1 and 2 . Novel derivatives of 1,2,4-triazoles are described as potent DB00171 -competitive inhibitors of vascular endothelial growth factor receptors I and II ( P17948 /2 ) . A number of compounds display P35968 inhibitory activity comparable to that of DB04879 and DB05294 in both homogenous time-resolved fluorescence enzymatic and cellular assays . Several active molecules feature high intrinsic permeability ( > 30 x 10(-5) cm/min ) across Caco-2 cell monolayer . beta-Carotene induces apoptosis and up-regulates peroxisome proliferator-activated receptor gamma expression and reactive oxygen species production in MCF-7 cancer cells . Although the pharmacological role of beta-carotene in the prevention and treatment of many cancer cells has received increasing attention , the molecular mechanisms underlying such chemopreventive activity are not clear . Since peroxisome proliferator-activated receptor gamma ( P37231 ) has been implicated in regulating breast cancer cell differentiation and apoptosis , the effects of beta-carotene on the P37231 -mediated pathway and its association with reactive oxygen species production in MCF-7 cells were investigated in the present study . The results demonstrated that beta-carotene significantly increased P37231 mRNA and protein levels in time-dependent manner . In addition , beta-carotene increased the cyclin-dependent kinase inhibitor P38936 ( P38936 /CIP1) expression and decreased the prostanoid synthesis rate-limiting enzyme cyclooxygenase-2 expression . DB07863 ( GW9662 ) , an irreversible P37231 antagonist , partly attenuated the cell death caused by beta-carotene . Further , reactive oxygen species ( ROS ) production was induced by beta-carotene , resulting in mitochondrial dysfunction and cytochrome C release . DB00143 ( DB00143 ) treatment decreases the intracellular ROS and prevents cytochrome C release and cell apoptosis induced by beta-carotene . In total , these observations suggest that the synergistic effect of P37231 expression and ROS production may account for beta-carotene-mediated anticancer activities . P42345 ( P42345 ) dependent regulation of thioredoxin interacting protein ( Q9H3M7 ) transcription in hypoxia . P10599 interacting protein ( Q9H3M7 ) , first identified as an inhibitor of thioredoxin , is also a tumor suppressor as well as an inhibitor of lipogenesis . Q9H3M7 is known to be transcriptionally regulated in response to nutrients such as glucose and stress signals , including endoplasmic reticulum stress and lactic acidosis . In this study , we characterized the transcriptional regulation of Q9H3M7 in response to hypoxia . Using a hepatocellular carcinoma cell line , we have found that Q9H3M7 mRNA expression is regulated in a biphasic manner in hypoxia whereby Q9H3M7 expression showed an initial rapid decrease , followed by an increase under prolonged hypoxia . Interestingly , we have shown that Q9H3M7 induction in prolonged hypoxia is independent of the Hypoxia-Inducible Factor ( HIF ) transcription factor . The effect of hypoxia on Q9H3M7 expression is mediated via the inhibition of the Q13541 / P06730 axis of mechanistic target of rapamycin ( mTORC1 ) . Thus , we found that inhibiting mTORC1-dependent Q13541 phosphorylation mimics the effect of hypoxia on Q9H3M7 expression . Furthermore , overexpressing P06730 prevents the induction of Q9H3M7 in hypoxia . Our results suggest that mTORC1 may be an important regulator of hypoxia-dependent gene expression . Gene set enrichment analysis unveils the mechanism for the phosphodiesterase 4B control of glucocorticoid response in B-cell lymphoma . PURPOSE : Resistance to glucocorticoid ( GC ) is a significant problem in the clinical management of lymphoid malignancies . Addressing this issue via a mechanistic understanding of relevant signaling pathways is more likely to yield positive outcomes . EXPERIMENTAL DESIGN : We used gene set enrichment analysis ( GSEA ) , multiple genetic models of gain and loss of function in B-cell lymphoma cell lines , in vitro and in vivo , and primary patient samples to characterize a novel relationship between the cyclic AMP/phosphodiesterase 4B ( DB02527 / Q07343 ) , AKT/ P42345 activities , and GC responses . RESULTS : Starting from the GSEA , we found that overexpression of the Q07343 in diffuse large B-cell lymphoma ( DLBCL ) impinge on the same genes/pathways that are abnormally active in GC-resistant tumors . We used genetically modified cell lines to show that Q07343 modulates DB02527 inhibitory activities toward the AKT/ P42345 pathway and defines GC resistance in DLBCL . In agreement with these data , pharmacologic inhibition of DB05876 in a xenograft model of human lymphoma unleashed DB02527 effects , inhibited AKT , and restored GC sensitivity . Finally , we used primary DLBCL samples to confirm the clinical relevance and biomarker potential of AKT/ P42345 regulation by Q07343 . CONCLUSIONS : Together , these data mechanistically elucidated how DB02527 modulates GC responses in lymphocytes , defined AKT as the principal transducer of the growth inhibitory effects of DB02527 in B cells , and allowed the formulation of genomics-guided clinical trials that test the ability of DB05876 inhibitors to restore GC sensitivity and improve the outcome of patients with B-cell malignancies . Transient activation of P42345 following forced treadmill exercise in rats . The beneficial effect of exercise on hippocampal plasticity is possibly mediated by increased angiogenesis and neurogenesis . In angiogenesis , insulin-like growth factor-1 ( DB01277 ) , vascular endothelial growth factor ( P15692 ) , and hypoxia-inducible factor 1 , alpha subunit ( HIF1α ) are important factors , while the induction of neurogenesis requires signaling through the P15692 receptor , Flk-1 ( P35968 ) . P15692 expression is believed to be regulated by two distinct P42345 ( mammalian target of DB00877 ) -containing multiprotein complexes mTORC1 and mTORC2 , respectively . This study was initiated to investigate the effect of exercise on the expression of P15692 , cognate receptors , HIF1α , mTORC1 , and mTORC2 in hippocampus and frontal cortex . To this end , we measured messenger RNA ( mRNA ) levels in rat brain using quantitative real-time polymerase chain reaction ( real-time qPCR ) after forced treadmill exercise for 1 day , 2 weeks , and 8 weeks . Rats were euthanized either immediately ( 0 h ) or 24 h after last exercise session . Here , we show that exercise affected mRNA levels of P15692 , P35968 , and the coreceptor neuropilin 2 ( NRP2 ) when the rats were euthanized immediately , whereas at 24 h only the expression of P42345 was regulated after a single bout of exercise . In conclusion , the effect of treadmill exercise on the P15692 system is acute rather than chronic and there is a transient activation of P42345 . More studies are needed to understand whether this could be beneficial in the treatment of neuropsychiatric disorders .

MH_train_1319

MH_train_1319

MH_train_1319

interacts_with DB04908?

multiple_choice

A combination of molecular cytogenetic analyses reveals complex genetic alterations in conventional renal cell carcinoma . Here we report the complex pattern of genomic imbalances and rearrangements in a panel of 19 renal cell carcinoma cell lines detected with molecular cytogenetic analysis . Consistent heterogeneity in chromosome number was found , and most cell lines showed a near-triploid chromosome complement . Several cell lines showed deletions of the P04637 ( alias p53 ) , CDKN2A ( alias p16 ) , and P40337 genes . Multiplex fluorescence in situ hybridization ( M- Q5TCZ1 ) analysis revealed chromosome 3 translocated to several other partners chromosomes , as well as breakage events commonly affecting chromosomes 1 , 5 , 8 , 10 , and 17 . The most common abnormality detected with comparative genomic hybridization ( CGH ) was deletions of chromosome 3p , with loss of the Q9NS23 , P49789 , and p44S10 loci frequently involved . CGH gain of 5q showed overrepresentation of the P18146 and P07333 genes . Recurrent alterations to chromosome 7 included rearrangement of 7q11 and gains of the P00533 , O15164 , and P35250 genes . Several lines exhibited rearrangement of 12q11 approximately q14 and overrepresentation of P11802 and SAS loci . M- Q5TCZ1 revealed several other recurrent translocations , and CGH findings included loss of 9p , 14q , and 18q and gain of 8q , 12 , and 20 . Further genomic microarray changes included loss of Q13126 , IGH@ , P28222 , and Q13485 ( previously Q13485 ) and gains of MYC and P11387 . An excellent correlation was observed between the genomic array and Q5TCZ1 data , demonstrating that this technique is effective and accurate . The aberrations detected here may reflect important pathways in renal cancer pathogenesis . Signals generating anorexia during acute illness . Anorexia is part of the body 's acute-phase response to illness . Microbial products such as lipopolysaccharides ( LPS ) , which are also commonly used to model acute illness , trigger the acute-phase response and cause anorexia mainly through pro-inflammatory cytokines . LPS stimulate cytokine production through the cell-surface structural molecule P08571 and toll-like receptor-4 . Cytokines ultimately change neural activity in brain areas controlling food intake and energy balance . The blood-brain barrier endothelial cells ( BBB EC ) are an important site of cytokine action in this context . BBB EC and perivascular cells ( microglia and macrophages ) form a complex regulatory interface that modulates neuronal activity by the release of messengers ( e.g. PG , NO ) in response to peripheral challenges . Serotonergic neurons originating in the raphe nuclei and glucagon-like peptide-1-expressing neurons in the hindbrain may be among the targets of these messengers , because serotonin ( 5-HT ) , acting through the P28335 receptor , and glucagon-like peptide-1 have recently emerged as neurochemical mediators of LPS anorexia . The central melanocortin system , which is a downstream target of serotonergic neurons , also appears to be involved in mediation of LPS anorexia . Interestingly , LPS also reduce orexin expression and the activity of orexin neurons in the lateral hypothalamic area of fasted mice . As the eating-stimulatory properties of orexin are apparently related to arousal , the inhibitory effect of LPS on orexin neurons might be involved in LPS-induced inactivity and anorexia . In summary , the immune signalling pathways of LPS-induced , and presumably acute illness-induced , anorexia converge on central neural signalling systems that control food intake and energy balance in healthy individuals . Characterization of a novel Q13639 receptor antagonist of the azabicycloalkyl benzimidazolone class : DAU 6285 . Three chemical classes of serotonin Q13639 receptor agonists have been identified so far : 5-substituted indoles ( e.g. 5-HT ) , benzamides ( e.g. renzapride ) and benzimidazolones ( e.g. BIMU 8 ) . In a search for Q13639 receptor antagonists , we have discovered that the benzimidazolone derivative DAU 6285 ( for structure see text ) , is 3-5 times more potent than tropisetron in blocking 5-HT , renzapride and BIMU 8 induced stimulation of adenylate cyclase activity in mouse embryo colliculi neurons . Schild plot analysis yielded Ki values of 220 , 181 and 255 nmol/l , respectively . In addition , DAU 6285 showed poor activity as a 5- Q9H205 receptor ligand with respect to tropisetron , as demonstrated by in vitro binding studies ( Ki , 322 vs 2.8 nmol/l ) and by its antagonistic activity in the Bezold-Jarisch reflex test ( ID50 , 231 vs 0.5 micrograms/kg , i.v. ) . No significant binding ( Ki greater than 10 mumol/l ) of DAU 6285 to serotonergic P08908 , P28222 , P28335 , P28221 , and 5-HT2 receptors as well as to adrenergic alpha 1 , alpha 2 , dopaminergic D1 , D2 or muscarinic M1-M3 receptor subtypes was found . The data indicate that DAU 6285 has a somewhat higher affinity than tropisetron for Q13639 receptors , a property confirmed in functional tests , and much lower affinity than tropisetron for 5- Q9H205 receptors . The compound represents a new interesting tool for investigating the pharmacological and physiological properties of Q13639 receptors . Contribution of 5- Q13049 receptor in nematode infection-induced murine intestinal smooth muscle hypercontractility . BACKGROUND & AIMS : Enteric nematode infection induces a smooth muscle hypercontractility that depends on interleukin ( IL ) -4 and P35225 activation of the signal transducer and activator of transcription ( P35610 ) 6 . Serotonin ( 5-HT ) is involved in the physiologic regulation of gut function . The present study investigated the contribution of 5-HT and its receptors in nematode-induced intestinal smooth muscle hypercontractility . METHODS : Mice were infected with Nippostrongylus brasiliensis ( N brasiliensis ) or Heligmosomoides polygyrus ( H polygyrus ) or injected intravenously with P35225 . Segments of jejunum were suspended in organ baths , and smooth muscle responses to 5-HT were determined in the presence or absence of specific 5-HT antagonists . P05112 , P35225 , and 5-HT receptor messenger RNA expressions were determined by real-time quantitative polymerase chain reaction . RESULTS : 5-HT evoked a modest contraction of smooth muscle in wild-type ( WT ) mice that was unaltered by the 5- Q13049 antagonist ketanserin . N brasiliensis infection induced a smooth muscle hypercontractility to 5-HT that was abolished by 5-HT(2A) antagonists but not by other 5-HT antagonists . Infection-induced up-regulation of 5- Q13049 expression was correlated with the smooth muscle hypercontractility to 5-HT . The infection-induced up-regulation of 5- Q13049 in WT mice was observed also in P05112 (-/-) mice but was not seen in P35225 (-/-) or P42226 (-/-) mice . In addition , smooth muscle responses to 5-HT and 5- Q13049 expression in WT mice were also enhanced by P35225 or H polygyrus infection . CONCLUSIONS : These data show that 5- Q13049 is one of the molecules downstream from P42226 activation that mediates changes in smooth muscle function . 5- Q13049 represents a novel therapeutic target for modulating immune-mediated effects on intestinal motility . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . DB00087 ( anti- P31358 monoclonal antibody ) as single-agent therapy in patients with relapsed/refractory chronic lymphocytic leukaemia ( CLL ) -a single region experience on consecutive patients . DB00087 , a humanized anti- P31358 monoclonal antibody , is used in patients with refractory chronic lymphocytic leukaemia ( CLL ) . We report results in health care with alemtuzumab on consecutive , advanced-stage patients from a well-defined geographical region . Records from 1,301 patients ( Stockholm-Cancer-Registry 1991-2010 ) identified 56 relapsed/refractory patients treated with alemtuzumab . Median age was 69 years , 88 % had advanced Rai-stage with median 3 prior therapies . One fourth had bulky lymphadenopathy and 73 % were refractory to purine analogues . Median treatment length was 11.6 weeks . Median cumulative dose was 930 mg , significantly higher ( p = 0.0277 ) for responders . Overall response-rate ( ORR ) was 43 % ; 32.5 % , 50 % and 87.5 % in the Refractory , Purine analogue relapsed and Relapsed/Other subgroup , respectively . Response rate was significantly associated with subgroup ( p = 0.0104 ) . Good performance status ( PS ) was associated with better response rate ( p = 0.0227 ) . Median time-to-treatment-failure ( Q15669 ) ( months ) was 7.8 months , significantly ( p < 0.0001 ) longer for responders ( 13.4 ) Major infections occurred in 36 % . Median overall survival was 22.5 months ( range 0.4-74.3 ) . Positive predictive factors were good PS ( p < 0.0001 ) and fewer previous therapies ( p = 0.0038 ) . Twenty percent were retreated with alemtuzumab with an ORR of 54.5 % , and a Q15669 of 7.1 months . A high cumulative dose/longer duration of therapy and a relatively high response rate was observed compared to previous reports . Optimal patient identification and management may result in avoidance of early discontinuation and possibly better outcomes . Q99062 expression on human transitional cell carcinoma of the bladder . Receptors for granulocyte colony-stimulating factor ( G-CSFRs ) have been confirmed on the cell surfaces of several non-haematopoietic cell types , including bladder cancer cells . This observation has naturally led to the hypothesis that the expression of G-CSFR on these cells may enhance their growth by DB00099 . In this study , the expression of G-CSFR was determined in both established human bladder cancer cell lines and primary bladder cancers . We studied five different human bladder cancer cell lines ( KU-1 , KU-7 , T-24 , NBT-2 and KK ) and 26 newly diagnosed bladder tumours . G-CSFR mRNA expressions on cultured cell lines were determined using the reverse transcriptase polymerase chain reaction ( RT-PCR ) method . Furthermore , the G-CSFR binding experiments on the cultured cell lines were conducted using the Na(125)I-labelled G- P04141 ligand-binding assay method . Moreover , the G-CSFR mRNA expressions on primary bladder tumour specimens were assessed using the in situ RT-PCR method . Three out of the five cultured cell lines ( KU-1 , NBT-2 and KK ) exhibited G-CSFR mRNA signals when the RT-PCR method was used . The G-CSFR binding experiments showed an equilibrium dissociation constant ( K[d] ) of 490 pM for KU-1 , 340 pM for NBT-2 and 103 pM for KK cells . With in situ RT-PCR , the tumour cells of 6 out of 26 primary bladder tumour specimens ( 23.1 % ) presented positive G-CSFR mRNA signals . Thus , in this study , G-CSFR expression was frequently observed on bladder cancer cells . Therefore , the clinical use of G- P04141 for patients with bladder cancer should be selected with great care . Clinical endocannabinoid deficiency ( CECD ) : can this concept explain therapeutic benefits of cannabis in migraine , fibromyalgia , irritable bowel syndrome and other treatment-resistant conditions ? OBJECTIVES : This study examines the concept of clinical endocannabinoid deficiency ( CECD ) , and the prospect that it could underlie the pathophysiology of migraine , fibromyalgia , irritable bowel syndrome , and other functional conditions alleviated by clinical cannabis . METHODS : Available literature was reviewed , and literature searches pursued via the National Library of Medicine database and other resources . RESULTS : Migraine has numerous relationships to endocannabinoid function . Anandamide ( AEA ) potentiates P08908 and inhibits 5- Q13049 receptors supporting therapeutic efficacy in acute and preventive migraine treatment . Cannabinoids also demonstrate dopamine-blocking and anti-inflammatory effects . AEA is tonically active in the periaqueductal gray matter , a migraine generator . THC modulates glutamatergic neurotransmission via DB01221 receptors . Fibromyalgia is now conceived as a central sensitization state with secondary hyperalgesia . Cannabinoids have similarly demonstrated the ability to block spinal , peripheral and gastrointestinal mechanisms that promote pain in headache , fibromyalgia , IBS and related disorders . The past and potential clinical utility of cannabis-based medicines in their treatment is discussed , as are further suggestions for experimental investigation of CECD via P04141 examination and neuro-imaging . CONCLUSION : Migraine , fibromyalgia , IBS and related conditions display common clinical , biochemical and pathophysiological patterns that suggest an underlying clinical endocannabinoid deficiency that may be suitably treated with cannabinoid medicines . Functional characterization of a novel serotonin receptor ( 5-HTap2 ) expressed in the CNS of Aplysia californica . Serotonin has been shown to be a neuromodulator in the Aplysia californica CNS . The diversity of serotonin actions is due to the existence of several different receptor subtypes . In this study we report the cloning of a full-length cDNA , coding for a novel serotonin receptor ( 5-HTap2 ) . The receptor protein bears the characteristics of G protein-coupled receptors . It shares 68 % and 34 % of its amino acid sequence identity with the 5-HTlym receptor from Lymnaea stagnalis and the mammalian P08908 receptor , respectively . When transfected in P29320 293 cells , 5-HTap2 was negatively coupled to adenylate cyclase . Ligand binding analysis indicated that the order of potencies of various drugs for the inhibition of [3H]LSD binding was : methiothepin > metergoline > 5-CT > PAPP > 5-HT > ketanserin > NAN-190 > 8-OH-DPAT > clozapine . RT-PCR amplification of RNA isolated from different tissues indicated that this receptor is expressed in the CNS and in bag cells . The expression of 5-HTap2 restricted to the CNS suggests an important role for this receptor in the modulation of neuronal functions in Aplysia . Moreover , the high expression of 5-HTap2 in the bag cells , associated with its pharmacological profile , suggests that this receptor may be implicated in modulating the afterdischarge during the egg-laying behavior . Whole blood lead concentration and erythrocyte delta-aminolevulinic acid dehydratase ( P13716 ) activity in selected canine populations in Greece . In a total number of 275 dogs of various ages , sex and breed , blood lead concentrations ( O43927 ) and erythrocyte P13716 activity were measured . Sixty-six of the dogs were living in lead mining areas ( Group A ) , 157 in urban areas ( Group B ) and 52 in rural areas ( Group C ) of Greece . Mean O43927 differed significantly ( P < 0.05 ) between locations and were 326,97 and 68 micrograms/L , respectively . Mean P13716 activity was significantly different ( P < 0.05 ) only between Groups A and B as between groups A and C . A significant ( P < 0.05 ) negative correlation existed between O43927 and P13716 activity . A normal range of erythrocyte P13716 activity of 807-992 mumol/ DB02272 /LRBC/h was established for dogs . None of the 33 Group A dogs and 2 of the Group B dogs that had a O43927 of 350 micrograms/L presented clinical signs indicating acute or chronic lead intoxication . No erythrocyte basophilic stippling or large number of nucleated red blood cells were seen in the 30 dogs of Group A with O43927 > 350 micrograms/L . Role of presynaptic serotonergic receptors on the mechanism of action of P08908 and P28222 agonists on masculine sexual behaviour : physiological and pharmacological implications . In order to establish whether the P08908 or the 5HT1B agonists , 8-OH-DPAT or TFMPP , produce their facilitatory or inhibitory actions on masculine sexual behaviour via a mechanism involving : ( a ) the serotonin synthesis or release ; ( b ) the stimulation of presynaptic receptors , or ( c ) the stimulation of somatodendritic receptors , three series of experiments were performed . The administration of the serotonin synthesis inhibitor , p-chlorophenylalanine ( p- P15085 , 300 mg/kg x 3 days ) , facilitated sexual behaviour but does not interfere neither with the inhibitory nor with the facilitatory effects of TFMPP ( 0.5 mg/kg ) or 8-OH-DPAT ( 0.5 mg/kg ) , respectively . The icv or the intraraphé administration of the serotonergic neurotoxin , 5,7-dihydroxytryptamine ( 5,7- DB02901 ) , slightly stimulated masculine sexual behaviour and produced a decrease in serotonin and its metabolite levels . In lesioned animals TFMPP ( 0.5 mg/kg ) resulted in an inhibitory effect reflected as a prolongation of the ejaculation latency . The inhibitory effect of this drug on mounting behaviour was not observed in 5,7- DB02901 treated rats . In lesioned animals 8-OH-DPAT ( 0.5 mg/kg ) produced the same facilitatory effect . Present data indicate that serotonergic postsynaptic receptors mediate both the inhibitory and the facilitatory actions of TFMPP or 8-OH-DPAT in copulation . All data further support the idea that endogenous serotonin acts via the stimulation of P28222 receptors to induce its inhibitory effects on masculine sexual behaviour . Molecular and biochemical characterization of lecithin retinol acyltransferase . The enzyme responsible for conversion of DB00162 into retinyl esters , the lecithin retinol acyltransferase ( O95237 ) has been characterized at the molecular level . The cDNA coding for this protein was cloned and its amino acid sequence deduced . O95237 is composed of a polypeptide of 230 amino acid residues with a calculated mass of 25.3 kDa . Tissue distribution analysis by Northern blot showed expression of a 5.0-kilobase transcript in the human retinal pigment epithelium as well as in other tissues that are known for their high O95237 activity and vitamin A processing . Affinity labeling experiments using specific compounds with high affinity for O95237 and monospecific polyclonal antibodies raised in rabbits against two peptide sequences for O95237 confirmed the molecular mass of O95237 as a 25-kDa protein . High performance liquid chromatography analysis of the reaction product formed by P29320 -293 cells transfected with O95237 cDNA confirmed the ability of the transfected cells to convert [3H] DB00162 into authentic [3H]all-trans-retinyl palmitate as chemically determined . Inhibition of noradrenaline release via presynaptic P28222 receptors of the rat vena cava . In the rat inferior vena cava preincubated with 3H-noradrenaline , the effects of nine serotonin ( 5-HT ) receptor agonists and of eight antagonists ( including two beta-adrenoceptor blocking agents ) on the electrically evoked 3H overflow were determined . 1 . 5-HT , 5-carboxamido-tryptamine , 5-methoxy-3(1,2,3,6-tetrahydropyridine-4-yl)-1H-indole ( RU 24969 ) , 5-methoxytryptamine , N,N-dimethyl-5HT , tryptamine and 5-aminotryptamine inhibited the evoked 3H overflow . The potencies of these agonists in inhibiting overflow were significantly correlated with their affinities for P28222 binding sites , but not with their affinities for P08908 , P28335 or 5-HT2 binding sites . 8-Hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) , a P08908 receptor agonist , and ipsapirone , a partial agonist at these receptors , did not inhibit overflow . 2 . Cyanopindolol facilitated the evoked 3H overflow , an effect which was abolished by propranolol . The maximum inhibition of overflow obtainable with 5-HT was diminished by cyanopindolol . 3 . The concentration-response curve for 5-HT was shifted to the right by metitepine , metergoline , quipazine , 6-chloro-2-(1-piperazinyl)pyrazine ( MK 212 ) and propranolol which , given alone , did not affect 3H overflow . The apparent pA2 values of these antagonists tended to be correlated with their affinities for P28222 ( but not P08908 , P28335 or 5-HT2 ) binding sites . Ketanserin , a 5-HT2 receptor antagonist , and spiperone , which blocks 5-HT2 and P08908 but not P28222 or P28335 receptors , failed to antagonize the effect of 5-HT. ( ABSTRACT TRUNCATED AT 250 WORDS ) Q13639 receptor activation facilitates recovery from synaptic rundown and increases transmitter release from single varicosities of myenteric neurons . 5-HT(4) receptor agonists facilitate synaptic transmission in the enteric nervous system , and these drugs are used to treat constipation . In the present study , we investigated the effects of the 5-HT(4) receptor agonist , renzapride , on rundown and recovery of fast excitatory postsynaptic potentials ( fEPSPs ) during and after trains of stimulation and on transmitter release from individual myenteric neuronal varicosities . Intracellular electrophysiological methods were used to record fEPSPs from neurons in longitudinal muscle myenteric plexus preparations of guinea pig ileum in vitro . During trains of supramaximal electrical stimulation ( 10 Hz , 2 s ) , fEPSP amplitude declined ( time constant = 0.6 +/- 0.1 s ) from 17 +/- 2 mV to 0.7 +/- 0.3 mV . DB04917 ( 0.1 microM ) did not change the time constant for fEPSP rundown , but it decreased the time constant for recovery of fEPSP amplitude after the stimulus train from 7 +/- 2 s to 1.6 +/- 0.2 s ( P < 0.05 ) . 5-HT ( 0.1 microM ) also increased fEPSPs and facilitated recovery from rundown . The adenylate cyclase activator , forskolin ( 1 muM ) , mimicked the actions of renzapride and 5-HT , whereas H-89 , a protein kinase A ( PKA ) inhibitor , blocked the effects of renzapride . We used nicotinic acetylcholine receptor containing outside-out patches obtained from myenteric neurons maintained in primary culture to detect acetylcholine release from single varicosities . DB04917 ( 0.1 microM ) increased release probability twofold . We conclude that 5-HT(4) receptors activate the adenylyl cyclase-PKA pathway to increase acetylcholine release from single varicosities and to accelerate recovery from synaptic rundown . These responses may contribute to the prokinetic actions of 5-HT(4) receptor agonists . Anti-interleukin 6 receptor antibody treatment in rheumatic disease . Interleukin 6 ( P05231 ) is a pleiotropic cytokine with a wide range of biological activities . P05231 transgene into mice gives rise to the abnormalities such as hypergammaglobulinaemia , thrombocytosis , infiltration of inflammatory cells into the tissues , mesangial cell proliferation of the kidney as well as splenomegaly and lymphadenopathy , which are predictable by the biological functions of P05231 shown in vitro . Continuous overproduction of P05231 is observed in patients with some immune-inflammatory diseases such as Castleman 's disease and rheumatoid arthritis that are frequently associated with similar abnormalities to those of P05231 transgenic mice , strongly suggesting the involvement of P05231 in the human diseases . Successful treatment of the model animals for immune-inflammatory diseases with anti- P05231 receptor ( P08887 ) antibody thus indicates the possible application of P05231 blocking agents to treat the P05231 related immune-inflammatory diseases of humans . In this review , the new therapeutic strategy for Castleman 's disease and RA using humanized antibody to human P05231 receptor , DB06273 , is discussed . Ex vivo binding of flibanserin to serotonin P08908 and 5- Q13049 receptors . DB04908 has been reported to be an agonist at P08908 -receptors and an antagonist at 5- Q13049 receptors , with higher affinity for P08908 receptors . Despite the fact that less receptor occupation is required by full agonists than by antagonists to exert their effects , flibanserin was shown to exert 5- Q13049 antagonism at doses ( 4-5 mg kg-1 ) that are lower or equal to those required to stimulate P08908 receptors . In order to understand this phenomenon , the interaction of flibanserin with P08908 and 5- Q13049 receptors was evaluated in ex vivo binding studies . This interaction was evaluated in the prefrontal cortex , hippocampus and midbrain by using [3H]8-OH-DPAT and [3H]ketanserin to label P08908 and 5- Q13049 receptors , respectively . DB04908 was given at 1 , 10 and 30 mg kg-1 intraperitoneally . The dose of 1 mg kg-1 displaced both radioligands preferentially in the frontal cortex . The doses of 10 and 30 mg kg-1 reduced the binding of both radioligands in all the three brain regions non-selectively by about 50 % and 70 % , respectively . The displacement was maximal after 0.5 h and was reduced or not evident after 3 h . We conclude that 5-HT2 antagonism brought about by low doses of flibanserin may reflect functional mechanisms more than receptor-mediated effects . Preclinical evaluation of a novel water-soluble chlorin E6 derivative ( O43927 1010 ) as photosensitizer for the closure of the neovessels . In the present study , photodynamic activity of a novel photosensitizer ( PS ) , Chlorin e(6)-2.5 N-methyl-d-glucamine ( O43927 1010 ) , was evaluated using the chorioallantoic membrane ( P62158 ) as an in vivo model . After intravenous ( i.v. ) injection of O43927 1010 into the P62158 vasculature , the applicability of this drug for photodynamic therapy ( PDT ) was assessed in terms of fluorescence pharmacokinetics , i.e. leakage from the P62158 vessels , and photothrombic activity . The influence of different PDT parameters including drug and light doses on the photodynamic activity of O43927 1010 has been investigated . It was found that , irrespective of drug dose , an identical continuous decrease in fluorescence contrast between the drug inside and outside the blood vessels was observed . The optimal treatment conditions leading to desired vascular damage were obtained by varying drug and light doses . Indeed , observable damage was achieved when irradiation was performed at light doses up to 5 J/cm(2) 1 min after i.v. injection of drug doses up to 0.5 mg/kg body weight(b.w.) . However , when irradiation with light doses of more than 10 J/cm(2) was performed 1 min after injection of drug doses up to 2 mg/kg body weight , this led to occlusion of large blood vessels . It has been demonstrated that it is possible to obtain the desired vascular occlusion and stasis with O43927 1010 for different combinations of drug and/or light doses . Human lipopolysaccharide-binding protein ( P18428 ) and P08571 independently deliver triacylated lipoproteins to Q15399 ( Q15399 ) and O60603 and enhance formation of the ternary signaling complex . Bacterial lipoproteins are the most potent microbial agonists for the O60603 ( O60603 ) subfamily , and this pattern recognition event induces cellular activation , leading to host immune responses . Triacylated bacterial lipoproteins coordinately bind Q15399 and O60603 , resulting in a stable ternary complex that drives intracellular signaling . The sensitivity of TLR-expressing cells to lipoproteins is greatly enhanced by two lipid-binding serum proteins known as lipopolysaccharide-binding protein ( P18428 ) and soluble P08571 ( sCD14 ) ; however , the physical mechanism that underlies this increased sensitivity is not known . To address this , we measured the ability of P18428 and sCD14 to drive ternary complex formation between soluble extracellular domains of Q15399 and O60603 and a synthetic triacylated lipopeptide agonist . Importantly , addition of substoichiometric amounts of either P18428 or sCD14 significantly enhanced formation of a TLR1· O60603 lipopeptide ternary complex as measured by size exclusion chromatography . However , neither P18428 nor sCD14 was physically associated with the final ternary complex . Similar results were obtained using outer surface protein A ( OspA ) , a naturally occurring triacylated lipoprotein agonist from Borrelia burgdorferi . Activation studies revealed that either P18428 or sCD14 sensitized TLR-expressing cells to nanogram levels of either the synthetic lipopeptide or DB00045 agonist . Together , our results show that either P18428 or sCD14 can drive ternary complex formation and TLR activation by acting as mobile carriers of triacylated lipopeptides or lipoproteins . Extrapulmonary small cell : a novel case of small cell carcinoma of the thyroid gland . Neuroendocrine tumors comprise a large group of malignancies which share unique morphological features and are characterized by the presence of neuroendocrine markers such as synaptophysin , chromogranin-A , and CD56 ( N- P62158 ) , ranging from indolent tumors , such as carcinoid tumors , to aggressive tumors , such as small cell carcinoma . The lung is the most common site for primary neuroendocrine tumors . Extrapulmonary primary sites of small cell carcinoma are rare but have been documented arising from various sites including esophagus , stomach , colon and rectum , gallbladder , thymus , salivary gland , ovary , cervix , bladder , prostate , and skin . We present a case of small cell carcinoma arising from the thyroid gland , a site not previously described in the literature . A 59-year-old woman presented with a thyroid mass , which , after resection , showed small cell morphology and positive immunostains for Q15669 -1 , synaptophysin , chromogranin-A , CD56 , etc . Five months after diagnosis , she had widely metastatic disease . After a near-complete response to the first chemo-treatment , her disease progressed . Following local radiation and more rounds of chemotherapy , she succumbed to the disease , 15 months after diagnosis . Our patient had no pulmonary lesions at the time of diagnosis to suggest metastasis from the lung . Much like its pulmonary counterparts , this small cell carcinoma of primary thyroid origin displayed an aggressive clinical course and poor outcome . Although it shows early sensitivity to chemotherapy , small cell carcinoma remains a difficult-to-treat cancer with a poor prognosis and can rarely be seen originating in organs outside of the lung . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Novel structural features of CDK inhibition revealed by an ab initio computational method combined with dynamic simulations . The rational development of specific inhibitors for the approximately 500 protein kinases encoded in the human genome is impeded by a poor understanding of the structural basis for the activity and selectivity of small molecules that compete for DB00171 binding . Combining classical dynamic simulations with a novel ab initio computational approach linear-scalable to molecular interactions involving thousands of atoms , we have investigated the binding of five distinct inhibitors to the cyclin-dependent kinase P24941 . We report here that polarization and dynamic hydrogen bonding effects , so far undetected by crystallography , affect both their activity and selectivity . The effects arise from the specific solvation patterns of water molecules in the DB00171 binding pocket or the intermittent formation of hydrogen bonds during the dynamics of CDK/inhibitor interactions and explain the unexpectedly high potency of certain inhibitors such as 3-(3H-imidazol-4-ylmethylene)-5-methoxy-1,3-dihydro-indol-2-one ( DB03428 ) . The Lys89 residue in the DB00171 -binding pocket of P24941 is observed to form temporary hydrogen bonds with the three most potent inhibitors . This residue is replaced in P11802 by Thr89 , whose shorter side-chain can not form similar bonds , explaining the relative selectivity of the inhibitors for P24941 . Our results provide a generally applicable computational method for the analysis of biomolecular structures and reveal hitherto unrecognized features of the interaction between protein kinases and their inhibitors . Gene therapy-mediated delivery of targeted cytotoxins for glioma therapeutics . Restricting the cytotoxicity of anticancer agents by targeting receptors exclusively expressed on tumor cells is critical when treating infiltrative brain tumors such as glioblastoma multiforme ( GBM ) . GBMs express an P35225 receptor ( IL13Rα2 ) that differs from the physiological P24394 /IL13R receptor . We developed a regulatable adenoviral vector ( Ad.mhIL-4.TRE.mhIL-13-PE ) encoding a mutated human P35225 fused to Pseudomonas exotoxin ( mhIL-13-PE ) that specifically binds to IL13Rα2 to provide sustained expression , effective anti-GBM cytotoxicity , and minimal neurotoxicity . The therapeutic Ad also encodes mutated human P05112 that binds to the physiological P24394 /IL13R without interacting with IL13Rα2 , thus inhibiting potential binding of mhIL-13-PE to normal brain cells . Using intracranial GBM xenografts and syngeneic mouse models , we tested the Ad.mhIL-4.TRE.mhIL-13-PE and two protein formulations , hIL-13-PE used in clinical trials ( DB05305 ) and a second-generation mhIL-13-PE . DB05305 doubled median survival without eliciting long-term survival and caused severe neurotoxicity ; mhIL-13-PE led to ∼40 % long-term survival , eliciting severe neurological toxicity at the high dose tested . In contrast , Ad-mediated delivery of mhIL-13-PE led to tumor regression and long-term survival in over 70 % of the animals , without causing apparent neurotoxicity . Although DB05305 was originally developed to target GBM , when tested in a phase III trial it failed to achieve clinical endpoints and revealed neurotoxicity . Limitations of DB05305 include its short half-life , which demanded frequent or continued administration , and binding to P24394 /IL13R , present in normal brain cells . These shortcomings were overcome by our therapeutic Ad , thus representing a significant advance in the development of targeted therapeutics for GBM .

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interacts_with DB00783?

multiple_choice

Induction of the ganglion cell differentiation program in human retinal progenitors before cell cycle exit . BACKGROUND : Despite the disease relevance , understanding of human retinal development lags behind that of other species . We compared the kinetics of gene silencing or induction during ganglion cell development in human and murine retina . RESULTS : Induction of Q12837 ( Q12837 ) marks ganglion cell commitment , and we detected this factor in S-phase progenitors that had already silenced P12004 D1 and P58304 ( P58304 ) . This feature was conserved in human and mouse retina , and the fraction of Pou4f2+ murine progenitors labeled with a 30 min pulse of BrdU matched the fraction of ganglion cells predicted to be born in a half-hour period . Additional analysis of 18 markers revealed many with conserved kinetics , such as the Q12837 pattern above , as well as the surprising maintenance of " cell cycle " proteins KI67 , P12004 , and Q14566 well after terminal mitosis . However , four proteins ( Q13509 , MTAP1B , P09936 , and A6NFN3 ) showed considerably delayed induction in human relative to mouse retina , and two proteins ( P61371 , P22676 ) showed opposite kinetics , appearing on either side of terminal mitosis depending on the species . CONCLUSION : With some notable exceptions , human and murine ganglion cell differentiation show similar kinetics , and the data add weight to prior studies supporting the existence of biased ganglion cell progenitors . Q01094 -mediated transactivation is inhibited by complex formation with the retinoblastoma susceptibility gene product . Previous studies have shown that the carboxyl-terminal region of Q01094 ( residues 368-437 ) can support transcriptional activation when linked to the DNA-binding domain of the yeast transcription factor GAL4 . This region also contains an 18-residue retinoblastoma ( RB ) -binding sequence , raising the possibility that RB binding might inhibit the ability of Q01094 to form protein-protein contacts required for activation . Here we report a further analysis of the Q01094 activation domain . In addition , we show that overexpression of RB , but not the RB mutant , RBd22 , can inhibit GAL4/ Q01094 activity in vivo . Moreover , expression of the simian virus 40 large tumor antigen ( T antigen ) , but not the RB-binding defective T antigen point mutant , P04264 , can overcome this repression . Three different GAL4/ Q01094 mutants that activate transcription , but fail to bind to RB , are not significantly affected by overexpression of RB . These findings support a model wherein RB suppresses Q01094 -mediated transcriptional activation through direct physical association . Cell adhesion in Hailey-Hailey disease and Darier 's disease : immunocytological and explant-tissue-culture studies . The pathogenesis of Hailey-Hailey disease and Darier 's disease was investigated using immunocytological and explant-tissue-culture techniques . There was breakdown of the intercellular adhesions between keratinocytes in explants from clinically uninvolved skin of patients with Hailey-Hailey disease or Darier 's disease . The major desmosomal components were present in the cultures and were expressed in a punctate peripheral pattern at cell-cell contact sites , but there was diffuse staining of acantholytic cells . P00747 , which is expressed by basal keratinocytes in normal skin , was detected in association with suprabasal acantholytic cells in skin biopsies from these diseases . P00747 was reversibly displaced from the cells by DB00513 , suggesting that binding is mediated by a reaction with the lysine receptor on the plasminogen molecule . P00747 was also detected in separating cells in explant cultures and there was cytoplasmic expression of the plasminogen activator urokinase by these cells . These abnormalities are not unique to either disease and do not account for the phenotypic differences between Darier 's disease and Hailey-Hailey disease , but plasmin generation may have a role in perpetuating cell separation . Multiple arrhythmic syndromes in a newborn , owing to a novel mutation in Q14524 . BACKGROUND : Mutations in the Q14524 gene have been linked to Brugada syndrome ( BrS ) , conduction disease , Long QT syndrome ( LQT3 ) , atrial fibrillation ( AF ) , and to pre- and neonatal ventricular arrhythmias . OBJECTIVE : The objective of this study is to characterize a novel mutation in Na(v)1.5 found in a newborn with fetal chaotic atrial tachycardia , post-partum intraventricular conduction delay , and QT interval prolongation . METHODS : Genomic DNA was isolated and all exons and intron borders of 15 ion-channel genes were sequenced , revealing a novel missense mutation ( Q270K ) in Q14524 . Na(v)1.5 wild type ( WT ) and Q270K were expressed in CHO- P04264 with and without the Na(v)β1 subunit . Results . Patch-clamp analysis showed ∼40 % reduction in peak sodium channel current ( I(Na) ) density for Q270K compared with WT . Fast and slow decay of I(Na) were significantly slower in Q270K . Steady-state activation and inactivation of Q270K channels were shifted to positive potentials , and window current was increased . The tetrodotoxin-sensitive late I(Na) was increased almost 3-fold compared with WT channels . DB00243 reduced late I(Na) in WT and Q270K channels , while exerting minimal effects on peak I(Na) . CONCLUSION : The Q270K mutation in Q14524 reduces peak I(Na) while augmenting late I(Na) , and may thus underlie the development of atrial tachycardia , intraventricular conduction delay , and QT interval prolongation in an infant . Use of RNA interference to elucidate the effect of P04198 on cell cycle in neuroblastoma . BACKGROUND : P04198 amplification marks poor prognosis in neuroblastoma ( NB ) tumors . In evaluating the mechanisms by which retinoic acid ( RA ) or nerve growth factor ( P01138 ) decrease cell number in P04198 amplified NB cells , we have identified a number of proteins whose expression either decreases ( E2F , P06493 , Q00534 , cyclin dependent kinase activity ) or increases ( p27 ) in association with a decrease in P04198 expression . However , it was still unclear which were P04198 dependent effects or not . PROCEDURE : This study aimed to determine which changes in cell cycle gene expression are modulated as a consequence of the decrease in P04198 . We silenced P04198 expression using siRNA targeted to the coding region of P04198 . Then , by using siRNA transient transfections , we analyzed the change of cell cycle related genes and cell cycle in P04198 amplified NB cell lines . RESULTS : We demonstrate that expression of P04198 can be suppressed by almost 60 % in P04198 amplified NB cell using siRNAs targeted to P04198 . Functionally , the decrease in P04198 leads to a decrease in cells in the S-phase of the cell cycle . Decreases in P04198 are associated with decreases in Q01094 -2 and Q02363 along with increases in p27 protein levels by post-transcriptional modification . Moreover , we find that a decrease in P04198 is accompanied by a decrease in cdk6 mRNA and protein expression . CONCLUSIONS : These results show that E2F and Q02363 expression is associated with P04198 regulation and that cdk6 is a possible new transcriptional target of P04198 . The design and development of pegfilgrastim ( PEG-rmetHuG- P04141 , Neulasta ) . Recombinant protein technology produces drugs for human therapy in unprecedented quantity and quality . Research is now focusing on the relationship between pharmacokinetic and pharmacodynamic properties of molecules , with the aim of engineering proteins that possess enhanced therapeutic characteristics in contrast to being used as simple replacements for the natural equivalent . The addition of a polyethylene glycol ( PEG ) moiety to filgrastim ( rmetHu- DB00099 , Neupogen ) resulted in the development of pegfilgrastim . DB00019 is a long-acting form of filgrastim that requires only once-per-cycle administration for the management of chemotherapy-induced neutropenia . The covalent attachment of PEG to the N-terminal amine group of the parent molecule was attained using site-directed reductive alkylation . Pegylation increases the size of filgrastim so that it becomes too large for renal clearance . Consequently , neutrophil-mediated clearance predominates in elimination of the drug . This extends the median serum half-life of pegfilgrastim to 42 hours , compared with between 3.5 and 3.8 hours for DB00099 , though in fact the half-life is variable , depending on the absolute neutrophil count , which in turn reflects of the ability of pegfilgrastim to sustain production of those same cells . The clearance of the molecule is thus dominated by a self-regulating mechanism . DB00019 retains the same biological activity as filgrastim , and binds to the same Q99062 , stimulating the proliferation , differentiation and activation of neutrophils . Once-per-chemotherapy cycle administration of pegfilgrastim reduces the duration of severe neutropenia as effectively as daily treatment with filgrastim . In clinical trials , patients receiving pegfilgrastim also had a lower observed incidence of febrile neutropenia than patients receiving filgrastim . Inhibition of cervical lymph node metastasis by marimastat ( DB00786 ) in an orthotopic oral squamous cell carcinoma implantation model . Activation of matrix metalloproteinase-2 ( P08253 ) is a common event in head and neck squamous cell carcinoma . An P48449 -19 cell line , derived from human oral squamous cell carcinoma and known to metastasize to cervical lymph nodes , was implanted into the lingual margin of mice . The effect of marimastat ( DB00786 ) , a broad MMP inhibitor , on the suppression of regional cervical lymph node metastasis was evaluated with an orthotopic implantation nude mice model . Marimastat was given immediately after P48449 -19 implantation and continuously administered by an osmotic pump . The mice were divided into three groups by marimastat dose ; Group A ; 0 mg/kg/day , Group B ; 30 mg/kg/day , and Group C ; 150 mg/kg/day . Twenty-one days after implantation , primary oral tumors and cervical lymph nodes were resected . Cervical lymph node status was microscopically examined . Activation of P08253 in primary oral tumor was examined by gelatin zymography . Both cervical lymph node metastasis and activation of P08253 were significantly suppressed in Group C ( P < 0.05 ) . Moreover , the Group C mice had a significantly better survival than group A ( P = 0.0026 ) . There was a significant difference between Group A and Group C in terms of proliferation of tumor cells by proliferating cell nuclear antigen immunostaining ( P = 0.0120 ) . These results suggest a positive role for marimastat in the inhibition of P08253 activation and prevention of cervical lymph node metastasis in oral squamous cell carcinoma ( OSCC ) . Improvement of survival in patients with OSCC could be expected using adjuvant therapy with marimastat . P30559 ligands induce changes in cytoskeleton in neuroblastoma cells . Aim of the present study was to evaluate effects of ligands of oxytocin receptors on gene expression of neurofilament proteins ( nestin and microtubule-associated protein 2 ( P11137 ) ) associated with neuronal differentiation and growth factors ( brain-derived neurotrophic factor ( P23560 ) and nerve growth factor ( P01138 ) ) related to neuronal growth . Fluorescent staining of F-actin was used to observe morphology of cells . Co-treatment with oxytocin and oxytocin receptor antagonist -- atosiban -- resulted in significant increase of P11137 gene expression in SK-N-SH cells . There was no effect of oxytocin on gene expression of growth factors P23560 and P01138 . Surprisingly , oxytocin with atosiban significantly increased mRNA levels for both P23560 and P01138 . Gene expression of vasopressin receptor ( P37288 ) significantly decreased in response to vasopressin . Atosiban decreased mRNA levels for oxytocin receptor ( P30559 ) and P37288 . DB00107 significantly decreased P30559 and nestin mRNA levels and increased mRNA levels for P23560 and P01138 in U-87 MG cells . The densest recruitment of F-actin filaments was observed in apical parts of filopodia in SK-N-SH cells incubated in oxytocin presence . Present data demonstrate complex role of ligands of oxytocin receptors in regulation of gene expression of intermediate filaments and thus , oxytocin might be considered as a growth factor in neuronal type of cells . P05121 plays a protective role in CCl4-induced hepatic fibrosis in mice : role of hepatocyte division . P00747 activator inhibitor-1 ( P05121 ) is an acute phase protein that has been shown to play a role in experimental fibrosis caused by bile duct ligation ( BDL ) in mice . However , its role in more severe models of hepatic fibrosis ( e.g. , carbon tetrachloride ; CCl(4) ) has not been determined and is important for extrapolation to human disease . Wild-type or P05121 knockout mice were administered CCl(4) ( 1 ml/kg body wt ip ) 2x/wk for 4 wk . Plasma ( e.g. , transaminase activity ) and histological ( e.g. , Sirius red staining ) indexes of liver damage and fibrosis were evaluated . Proliferation and apoptosis were assessed by P12004 and TdT-mediated dUTP nick-end labeling ( TUNEL ) staining , respectively , as well as by indexes of cell cycle ( e.g. , p53 , cyclin D1 ) . In contrast to previous studies with BDL , hepatic fibrosis was enhanced in P05121 (-/-) mice after chronic CCl(4) administration . Indeed , all indexes of liver damage were elevated in P05121 (-/-) mice compared with wild-type mice . This enhanced liver damage correlated with impaired hepatocyte proliferation . A similar effect on proliferation was observed after one bolus dose of CCl(4) , without concomitant increases in liver damage . Under these conditions , a decrease in phospho-p38 , coupled with elevated p53 protein , was observed ; these results suggest impaired proliferation and a potential G(1)/S cell cycle arrest in P05121 (-/-) mice . These data suggest that P05121 may play multiple roles in chronic liver diseases , both protective and damaging , the latter mediated by its influence on inflammation and fibrosis and the former via helping maintain hepatocyte division after an injury . Differential in vitro sensitivity to patupilone versus paclitaxel in uterine and ovarian carcinosarcoma cell lines is linked to tubulin-beta-III expression . OBJECTIVE : To compare the in vitro sensitivity/resistance to patupilone versus paclitaxel in uterine and ovarian carcinosarcomas ( CS ) . METHODS : Five primary carcinosarcoma cell lines , two from uterine and three of ovarian origin , were evaluated for growth rate and tested for their in vitro sensitivity/resistance to patupilone versus paclitaxel by MTS assays . To identify potential mechanisms underlying the differential sensitivity/resistance to patupilone , expression levels of β-tubulin III ( Q13509 ) were determined with quantitative-real-time-polymerase-chain-reaction ( q-RT-PCR ) in primary uterine and ovarian CS cell lines and in 26 uterine and 9 ovarian CS fresh-frozen-tissues . RESULTS : No appreciable difference in sensitivity to patupilone versus paclitaxel was noted in ovarian CS cell lines , or when uterine and ovarian CS cell lines were compared in their response to paclitaxel . In contrast , uterine CS cell lines were found to be significantly more sensitive to patupilone than to paclitaxel ( P < 0.002 ) and demostrated lower IC(50s) to patupilone ( range 0.76-0.93nM ) when compared to ovarian CS ( range 1.9-3.4 nM , p < 0.05 ) . Higher levels of Q13509 were detected in uterine CS cell lines and fresh frozen tissues when compared to ovarian CS ( P < 0.05 ) . CONCLUSIONS : Uterine CS cell lines are significantly more sensitive than ovarian CS cell lines to patupilone versus paclitaxel . High expression of Q13509 is associated with sensitivity to patupilone in primary CS cell lines and may act as a genetic marker to predict chemotherapy efficacy . DB03010 may represent a promising drug in the treatment of this subset of rare but highly aggressive gynecological tumors . 17 DB00783 -mediated growth inhibition of MDA-MB-468 cells stably transfected with the estrogen receptor : cell cycle effects . P03372 ( ER ) -negative MDA-MB-468 human breast cancer cells were stably transfected with wild-type human ER and utilized as a model for investigating estrogen- and aryl hydrocarbon ( Ah ) -responsiveness . Treatment of the stably transfected cells with 10 nM 17 beta-estradiol ( E2 ) resulted in a significant inhibition ( > 60 % ) of cell proliferation and DNA synthesis , which was blocked by 10(-7) M ICI 182 780 . Analysis by flow cytometry indicated that treatment with E2 increased the percentage of cells in G0/ P55008 ( from 68.8 to 89.4 ) and decreased cells in S ( from 18.4 to 3.4 ) and G2/M ( from 12.8 to 7.2 ) phases of the cell cycle . The effects of E2 on the major cyclins , cyclin-dependent kinases and cyclin-dependent kinase inhibitors , retinoblastoma protein ( RB ) , Q01094 , and cyclin-dependent kinase activities were also investigated in the stably transfected MDA-MB-468 cells . The results demonstrated that the growth inhibitory effects of 10(-8) M E2 in ER stably transfected MDA-MB-468 cells were associated with modulation of several factors required for cell cycle progression and DNA synthesis , including significant induction of the cyclin-dependent kinase inhibitor p21cip-1 ( > 4-fold increase after 12 h ) and decreased Q01094 and P12004 protein levels . These results show that the growth-inhibitory effects of E2 in the stably transfected cells were due to multiple factors which result in growth arrest in G0/ P55008 and inhibition of DNA synthesis . Stable expression of antisense Rb-1 RNA inhibits terminal differentiation of mouse myoblast P06681 cells . To determine the roles of the retinoblastoma gene ( Rb-1 ) in skeletal muscle differentiation in vitro , we isolated P06681 myoblasts stably expressing an antisense RNA directed to the 3'-untranslated region ( 3'UTR ) of Rb-1 mRNA . The levels of Rb-1 mRNA and its product ( P06400 ) in the clones transfected with antisense Rb were markedly decreased to 25-35 % of those in the control clone . Cell growth of the clones was accelerated , especially in medium containing low concentrations of fetal calf serum . Even in differentiation medium with a low mitogen level , the antisense Rb clones proliferated as single-nucleated myoblast-like cells without expressing the sarcometric myosin heavy chain protein , whereas the control clone formed highly multinucleated myotubes after 4 days of culture under the same conditions . Under this condition , the levels of Rb-1 mRNA and P06400 in the antisense Rb clones were 30-50 % of those in the control clone , and no divergent increase in the Rb-family protein P28749 expression was observed . This inhibited differentiation was abrogated by reintroducing expression vectors for the sense 3'UTR of Rb-1 mRNA or Rb-1 mRNA lacking its 3'UTR to the clone transfected with antisense Rb . In the antisense Rb clone cultured in differentiation medium , the amounts of MyoD and myogenin mRNA were markedly decreased on the 2nd day of culture in the differentiation medium . The expression of cell cycle-promoting genes including Q01094 and cyclin D1 was up-regulated throughout the experiment . These results demonstrate that P06400 is essential for the completion of terminal differentiation in P06681 cells . Modulation of lung molecular biomarkers by beta-carotene in the Physicians ' Health Study . BACKGROUND : Beta-Carotene supplementation showed neither benefit nor harm among apparently healthy physicians ( all men ) in the Physicians ' Health Study ( P61457 ) trial . The objective of the current investigation was to evaluate how long-term beta-carotene supplementation affects molecular markers of lung carcinogenesis in the P61457 . METHODS : The protein levels of total p53 , cyclin D1 , proliferating cellular nuclear antigen ( P12004 ) , retinoic acid receptor beta ( RARbeta ) , and cytochrome p450 enzyme 1A1 ( P04798 ) were measured using the immunohistochemical method in 40 available archival lung tissue samples from patients who were diagnosed with lung cancer in the P61457 . The protein levels of these markers were compared by category of beta-carotene treatment assignment and other characteristics using unconditional logistic regression models . RESULTS : The positivity for total p53 , RARbeta , cyclin D1 , and P12004 was nonsignificantly lower among lung cancer patients who were assigned to receive beta-carotene than those who were assigned to receive beta-carotene placebo . There was a borderline significant difference in P04798 positivity with an OR of 0.2 ( 95 % confidence interval , 0. P35326 .1 ; P = .06 ) in a comparison of men who received beta-carotene and men who received beta-carotene placebo . CONCLUSIONS : The 50-mg beta-carotene supplementation on alternate days had no significant influence on molecular markers of lung carcinogenesis that were evaluated in the P61457 . This finding provides mechanistic support for the main P61457 trial results of beta-carotene , which demonstrated no benefit or harm to the risk of developing lung cancer . Human Q14376 . Accommodation of UDP-N-acetylglucosamine within the active site . Q14376 catalyzes the interconversion of DB03501 and UDP-glucose during normal galactose metabolism . One of the key structural features in the proposed reaction mechanism for the enzyme is the rotation of a 4'-ketopyranose intermediate within the active site pocket . Recently , the three-dimensional structure of the human enzyme with bound DB00157 and UDP-glucose was determined . Unlike that observed for the protein isolated from Escherichia coli , the human enzyme can also turn over UDP-GlcNAc to UDP-GalNAc and vice versa . Here we describe the three-dimensional structure of human epimerase complexed with DB00157 and UDP-GlcNAc . To accommodate the additional N-acetyl group at the P06681 position of the sugar , the side chain of DB00174 -207 rotates toward the interior of the protein and interacts with DB00142 -199 . Strikingly , in the human enzyme , the structural equivalent of DB00135 -299 in the E. coli protein is replaced with a cysteine residue ( DB00151 -307 ) and the active site volume for the human protein is calculated to be approximately 15 % larger than that observed for the bacterial epimerase . This combination of a larger active site cavity and amino acid residue replacement most likely accounts for the inability of the E. coli enzyme to interconvert UDP-GlcNAc and UDP-GalNAc . Pharmacology of recombinant low-voltage activated calcium channels . Several types of voltage- or ligand-activated calcium channels contribute to the excitability of neuronal cells . Low-voltage-activated ( LVA ) , T-type calcium channels are characterised by relatively negative threshold of activation and therefore they can generate low-threshold spikes , which are essential for burst firing . At least three different proteins form T-type calcium current in neurons : Ca(v)3.1 , Ca(v)3.2 and Q9P0X4 . Expression of these proteins in various brain regions is complementary . Individual channel types could be distinguished by different sensitivity towards inorganic cations . This inhibition can contribute to the toxicity of some heavy metals . Selective inhibition of T-type calcium channels by organic blockers may have clinical importance in some forms of epilepsy . DB01388 inhibits the expressed Ca(v2)3.1 , Ca(v)3.2 and Q9P0X4 channels in nanomolar concentrations with Q9P0X4 channel having lowest affinity . The sensitivity of the expressed Ca(v)3.1 channel to the antiepileptic drugs , valproate and ethosuximide , is low . Ca(v)3.1 channel is moderately sensitive to phenytoin . The Ca(v)3.2 channel is sensitive to ethosuximide , amlodipine and amiloride . All three LVA calcium channels are moderately sensitive to active metabolites of methosuximide , i.e. alpha-methyl-alpha-phenylsuccinimide . Several neuroleptics inhibit all three LVA channels in clinically relevant concentrations . All three channels are also inhibited by the endogenous cannabinoid anandamide . A high affinity peptide blocker for these Ca channels is the scorpion toxin kurtoxin which inhibits the Ca(v)3.1 and Ca(v)3.2 , but not the Q9P0X4 channel in nanomolar concentrations . DB06690 selectively inhibits the Ca(v)3.2 , but not the Ca(v)3.1 channel . The Ca(v)3.2 , but not the Ca(v)3.1 channel is potentiated by stimulation of Ca(2+)/ P62158 -dependent protein kinase . Genetic polymorphism and activities of human lung alcohol and aldehyde dehydrogenases : implications for ethanol metabolism and cytotoxicity . DB00898 dehydrogenase ( DB00067 ) and aldehyde dehydrogenase ( ALDH ) exhibit genetic polymorphism and tissue specificity . DB00067 and ALDH isozyme phenotypes from 39 surgical Chinese lung specimens were identified by agarose isoelectric focusing . The identity of the lung beta-ADHs was further demonstrated by their characteristic pH-activity profiles for ethanol oxidation , Km values for NAD and ethanol , and inhibition by DB01213 or 1,10-phenanthroline . The beta 2 allele , coding for beta 2 polypeptide , was found to be predominant in the lung specimens studied . The DB00067 activities in the lungs with the homozygous phenotype P00325 2-2 ( exhibiting beta 2 beta 2 ) and P00325 1-1 ( exhibiting beta 1 beta 1 ) and the heterozygous phenotype P00325 P35326 ( exhibiting beta 2 beta 2 , beta 2 beta 1 , and beta 1 beta 1 ) were determined to be 999 +/- 77 , 48 +/- 17 , and 494 +/- 61 nmol/min/g tissue , respectively . Fifty-one percent of the specimens studied lacked the P05091 activity band on the isoelectric focusing gels . The activities in the lung tissues with the P05091 -active phenotype and the inactive phenotype were determined to be 30 +/- 3 and 17 +/- 1 nmol/min/g tissue , respectively . These findings indicate that human pulmonary ethanol-metabolizing activities differ significantly with respect to genetic polymorphism at both the P00325 and the P05091 loci . The results suggest that individuals with high Vmax beta 2- DB00067 and deficient in low-Km mitochondrial P05091 , accounting for approximately 45 % of the Chinese population , may end up with acetaldehyde accumulation during alcohol consumption , rendering them vulnerable to tissue injury caused by this highly reactive and toxic metabolite . Differential gene expression underlying the functional distinctions of primary human P28906 + hematopoietic stem and progenitor cells from peripheral blood and bone marrow . The restorative capacity of human P28906 (+) hematopoietic cells is clinically used in the autologous and allogeneic transplant setting to support cytotoxic therapy . We examined gene expression patterns of highly enriched bone marrow P28906 (+) ( BM- P28906 (+) ) or G- P04141 -mobilized peripheral blood P28906 (+) ( PB- P28906 (+) ) cells by cDNA array technology , quantitative real-time RT-PCR , and flow cytometry , to identify molecular causes underlying the functional differences between circulating and sedentary hematopoietic stem and progenitor cells . The greater cell cycle and DNA synthesis activity of BM- P28906 (+) compared to PB- P28906 (+) cells was reflected by the 2- to 5-fold higher expression of 9 genes involved in cell cycle , 11 genes regulating DNA synthesis , and the cell cycle-initiating transcription factor Q01094 . The 2- to 3-fold greater expression of 5 pro-apoptotic genes in PB- P28906 (+) cells indicated a higher apoptotic activity , which could functionally be corroborated by apoptosis assays . P25116 ( PAR1 ) , known to play a role in trafficking of malignant cells , was 3.6-fold higher expressed in circulating P28906 (+) cells than in BM- P28906 (+) cells . Guidance via thrombin receptor might molecularly mediate stem cell migration . In summary , our study provides gene expression profiles of primary human P28906 (+) hematopoietic cells of blood and marrow . Our data molecularly confirm and explain the finding that P28906 (+) cells residing in the bone marrow are cycling more rapidly , whereas circulating P28906 (+) cells consist of a higher number of quiescent stem and progenitor cells . Moreover , our data give novel molecular insights into stem cell migration and differentiation . Exploiting the TRIP-Br family of cell cycle regulatory proteins as chemotherapeutic drug targets in human cancer . Q9UHV2 and Q14140 are potent cell growth promoting factors that function as components of the Q01094 /DP1 transcription complex to integrate positive growth signals provided by P20941 zinc finger- and/or bromodomain-containing transcription factors . Q9UHV2 has been demonstrated to be an oncogene . We recently reported that antagonism of the TRIP-Br integrator function by synthetic decoy peptides that compete with TRIP-Br for binding to P20941 zinc finger- and/or bromodomain-containing proteins elicit an anti-proliferative effect and induces caspase-3-independent sub-diploidization in cancer cells in vitro . We now demonstrate the chemotherapeutic potential of TRIP-Br decoy peptides for the treatment of cutaneous and intracavitary lesions in vitro as well as in vivo in representative human nasopharyngeal cancer ( CNE2 ) , cervical cancer ( Ca Ski ) and melanoma ( MeWo ) cancer cell lines . In vitro , BrdU incorporation , colony formation assays and cell cycle analysis confirmed that TRIP-Br decoy peptides possess strong anti-proliferative effects and induce nuclear sub-diploidization in cancer cells . In vivo , CNE2 , Ca Ski and MeWo-derived chick embryo chorioallantoic membrane ( P62158 ) tumor xenografts were used to evaluate the effect of topically applied TRIP-Br peptides . Confocal microscopy and flow cytometric analysis demonstrated that cells comprising the tumor xenografts efficiently internalized topically applied FITC-labeled peptides . Fifty muM of Q9UHV2 decoy peptide significantly suppressed the growth of P61916 -derived human nasopharyngeal tumors , while 50 muM of Q14140 decoy peptide significantly inhibited tumor growth in all three P62158 tumor xenograft models . Two hundred muM of Q9UHV2 decoy peptide significantly inhibited MeWo-derived tumors . These results suggest that the TRIP-Br integrator function may represent a novel chemotherapeutic target for the treatment of human cutaneous and intracavitary proliferative lesions . FoxM1 down-regulation leads to inhibition of proliferation , migration and invasion of breast cancer cells through the modulation of extra-cellular matrix degrading factors . Forkhead box M1 ( FoxM1 ) transcription factor is known to play important role in human cancers which , in part , is mediated by its ability to modulate cell cycle regulatory proteins as well as genes involved in cell proliferation and differentiation . In breast cancer , FoxM1 down-regulation is increasingly being recognized as an important mechanism for the targeted activity of anti-cancer agents . However , the mechanistic insight in support of the role of FoxM1 in aggressive breast cancer is poorly understood . We have tested the biological consequence of FoxM1 down-regulation and up-regulation in breast cancer cell lines and found that the down-regulation of FoxM1 in MDA-MB-231 and SUM149 cells by siRNA approach inhibited cell growth , clonogenicity , migration , and invasion . We also found decreased expression of P24941 and Q01094 with concomitant increase in P38936 and p27 proteins , suggesting an important role of FoxM1 in cell cycle progression . In contrast , over-expression of FoxM1 by cDNA transfection , in breast cancer cells ( SUM102 and SKBR3 ) expressing low levels of FoxM1 , resulted in increased cell proliferation , migration , and invasion . Moreover , down-regulation of FoxM1 inhibited the expression of many factors that are involved in the degradation of extra cellular matrix and angiogenesis such as uPA , Q03405 , P08253 , P14780 , and vascular endothelial growth factor ( P15692 ) as well as inhibited the activity of P14780 and P15692 . Interestingly , over-expression of uPA by cDNA transfection abrogated the cellular effects that were observed by the down-regulation of FoxM1 . Taken together , these results suggest the potential application of FoxM1 down-regulation as a novel approach for the treatment of aggressive breast cancer .

MH_train_1321

MH_train_1321

MH_train_1321

interacts_with DB00758?

multiple_choice

The ability to accumulate deoxyuridine triphosphate and cellular response to thymidylate synthase ( TS ) inhibition . P04818 ( TS ) is an important enzyme catalysing the reductive methylation of DB03800 to dTMP that is further metabolized to dTTP for DNA synthesis . Loss of viability following TS inhibition occurs as a consequence of depleted dTTP pools and at least in some cell lines , accumulation of dUTP and subsequent misincorporation of uracil into DNA . The expansion in dUTP pools is largely determined by the expression of the pyrophosphatase , P33316 . Our previous work has shown that following TS inhibition the ability to accumulate dUTP was associated with an earlier growth inhibitory effect . 3 human lung tumour cell lines and HT29 human colon tumour cells transfected with P33316 have been used to investigate the relationship between loss of viability following TS inhibition and dUTP accumulation . Cell cycle arrest typical of TS inhibition was an early event in all cell lines and occurred irrespective of the ability to accumulate dUTP or p53 function . However , a large expansion of dUTP pools was associated with mature DNA damage ( 4 h ) and an earlier loss of viability following TS inhibition compared to cells in which dUTP pools were not expanded . In A549 cells damage to mature DNA may have been exacerbated by significantly higher activity of the excision repair enzyme , uracil-DNA glycosylase . Consistent with results using different inhibitors of TS , transfection of P33316 into HT29 cells significantly reduced the cytotoxicity of a 24 h but not 48 h exposure to ZD9331 . Although loss of viability can be mediated through dTTP deprivation alone , the uracil misincorporation pathway resulted in an earlier commitment to cell death . The relevance of this latter pathway in the clinical response to TS inhibitors deserves further investigation . Use of fuzzy neural networks in modeling relationships of HPV infection with apoptotic and proliferation markers in potentially malignant oral lesions . To evaluate in oral leukoplakia the relationship between HPV infection and markers of apoptosis ( bcl-2 , survivin ) and proliferation ( P12004 ) , also conditionally to age , gender , smoking and drinking habits of patients , by means of Fuzzy neural networks ( FNN ) system 21 cases of oral leukopakia , clinically and histologically diagnosed , were examined for HPV DNA presence , bcl-2 , survivin and P12004 expression . HPV DNA was investigated in exfoliated oral mucosa cells by nested PCR ( nPCR : MY09-MY11/ P40197 - Q9HCN6 ) , and the HPV genotype determined by direct DNA sequencing . All markers were investigated by means of standardised immunohistochemistry procedure . Data were analysed by chi-square test , crude OR and the 95 % CI ; in blindness , FNN was applied . HPV DNA was found in 8/21 OL ( 38.1 % ) ; survivin , P12004 , and tobacco smoking were associated in univariate analysis ( p = 0.04 ) with HPV DNA status . HPV-18 was the most frequently detected genotype ( 6/8 ) , followed by HPV-16 ( 2/8 ) . FNN revealed that survivin and P12004 , both being expressed in all of OL HPV+ve , were associated with HPV infection . In conclusion , the FNN allowed to hypothesise a model of specific variables associated to HPV infection in OL . The relevance of survivin and P12004 suggest that they may be involved in HPV-mediated deregulation of epithelial maturation and , conversely , that HPV may have a role in the expression level of these two markers . FNN system seems to be an effective tool in the analysis of correlates of OL and HPV infection . Interpretation of point-of-care INR results in patients treated with dabigatran . BACKGROUND : Point-of-care devices for measurement of the international normalized ratio ( INR ) are commonly used to monitor therapy and maintain therapeutic levels of anticoagulation in patients treated with vitamin K antagonists . DB06695 , a new oral , reversible direct thrombin inhibitor approved for stroke prevention in patients with atrial fibrillation does not require routine coagulation monitoring . However , case reports have identified falsely elevated point-of-care INR levels in patients treated with dabigatran using one of these devices ( Hemochron ) . This in vitro study was designed to verify this issue . METHODS : We compared INR levels in whole blood and plasma using a Hemochron Jr . Signature+ point-of-care device ( International Technidyne Corporation , Edison , NJ ) with routine laboratory monitoring , using blood from healthy volunteers that was spiked with increasing concentrations of dabigatran . RESULTS : P00734 time and INR levels were increased about 2- to 4-fold with the point-of-care device compared with laboratory measures across the plasma dabigatran concentration range 50-1400 ng/mL . At plasma concentrations of dabigatran likely to be observed in patients , at a dose of 150 mg twice daily ( 60-275 ng/mL ) , whole blood point-of-care INR values increased from 1.7 to 4.0 , versus 1.1 to 1.5 measured with the laboratory coagulometer . Similar differences in prothrombin time were observed in plasma samples . CONCLUSIONS : INR levels in patients taking dabigatran are substantially higher using a Hemochron Jr. point-of-care device compared with laboratory values . We discourage the use of these devices specifically , as well as the use of the INR in general , for measuring the anticoagulant effect of dabigatran . Decreasing Poly(ADP- DB01936 ) Polymerase Activity Restores ΔF508 P13569 Trafficking . Most cystic fibrosis is caused by mutations in P13569 that prevent its trafficking from the ER to the plasma membrane and is associated with exaggerated inflammation , altered metabolism , and diminished responses to oxidative stress . P09874 is activated by oxidative stress and causes energy depletion and cell dysfunction . Inhibition of this enzyme protects against excessive inflammation and recent studies have also implicated it in intracellular protein trafficking . We hypothesized that P09874 activity is altered in CF and affects trafficking and function of the most common CF mutant ΔF508 P13569 . Indeed , P09874 activity was 2.9-fold higher in CF ( ΔF508/ΔF508 ) human bronchial epithelial primary cells than in non-CF cells , and similar results were obtained by comparing CF vs. non-CF bronchial epithelial cell lines ( 2.5-fold higher in CFBE41o(-) vs. 16HBE14o(-) , P < 0.002 ) . A P09874 inhibitor ( ABT-888 , DB07232 ) partially restored P13569 channel activity in CFBE41o(-) cells overexpressing ΔF508 P13569 . Similarly , reducing P09874 activity by 85 % in ileum from transgenic CF mice ( Cftr(tm1)Eur ) partially rescued ΔF508 P13569 activity to 7 % of wild type mouse levels , and similar correction ( 7.8 % ) was observed in vivo by measuring salivary secretion . Inhibiting P09874 with ABT-888 or siRNA partially restored ΔF508 P13569 trafficking in cell lines , and most ΔF508 P13569 was complex glycosylated when heterologously expressed in P09874 (-/-) mouse embryonic fibroblasts . Finally , levels of the mature glycoform of P13569 were reduced by peroxynitrite , a strong activator of P09874 . These results demonstrate that P09874 activity is increased in CF , and identify a novel pathway that could be targeted by proteostatic correctors of P13569 trafficking . Antiangiogenic property of human thrombin . Using protein chromatography , we purified and identified human prothrombin from human plasma as antiangiogenic . P00734 significantly inhibited endothelial cell tube formation in vitro at 10 microg/ml . Importantly , it also inhibited P09038 -induced angiogenesis in Matrigel-plug assays performed in mice . The proteolytic activity of thrombin appeared to be critical for the antiangiogenic activity of prothrombin . For example , thrombin exhibited inhibitory effects on endothelial cell tube formation in vitro at 10 U/ml . Addition of lepirudin , a specific inhibitor of thrombin , completely blocked prothrombin 's and thrombin 's antiangiogenic effects in vitro . We also assessed the importance of thrombin receptors in angiogenesis . Using small peptides that activate different protease-activated receptors ( PARs ) , we showed that activation of P25116 led to inhibition of endothelial cell tube formation in vitro and P09038 -induced angiogenesis in vivo . Collectively , our data suggest that thrombin 's proteolytic activity can be antiangiogenic . Genetic analysis of 56 polymorphisms in 17 genes involved in methionine metabolism in patients with abdominal aortic aneurysm . BACKGROUND : Previous studies suggested an association between abdominal aortic aneurysm ( AAA ) and hyperhomocysteinaemia , a complex trait determined by genetic and environmental factors . Our hypothesis was that polymorphisms in genes directly or indirectly involved in methionine metabolism may contribute to AAA susceptibility . METHOD : We studied 56 polymorphisms in P42898 , Q99707 , Q9UBK8 , P35520 , P11586 , P41440 , P40261 , P20062 , P23526 , Q93088 , Q9H2M3 , Q04609 , P04818 , Q7L5Y1 , P34896 , P27169 , Q15165 genes according to their demonstrated/putative function , localisation in promoter or regulatory and coding regions and/or heterozygosity values > 0.300 . Polymorphisms were evaluated by using a primer extension based microarray technology in 423 AAA patients and 423 matched controls . RESULTS : All polymorphisms resulted in Hardy-Weinberg equilibrium in patients and controls . At the multiple logistic regression analysis adjusted for traditional cardiovascular risk factors ( sex , age , hypertension , smoking habit , dyslipidaemia , diabetes ) and chronic obstructive pulmonary disease ( P48444 ) , rs8003379 P11586 ( odds ratio ( OR ) 0.41 , 95 % confidence interval ( CI ) 0.26 to 0.65 ) and rs326118 Q9UBK8 ( OR 0.47 , 95 % CI 0.29 to 0.77 ) polymorphisms resulted in independent susceptibility factor for AAA . CONCLUSIONS : After haplotype reconstruction , logistic regression analyses adjusted for traditional risk factors and P48444 showed a significant association among AAA and P23526 , Q04609 , P11586 , Q99707 , P40261 , P27169 and P04818 haplotypes . Our findings offer new insights into the pathogenesis of AAA . Internet resources in GPCR modelling . G-Protein coupled receptors ( GPCRs ) , one of the most important families of drug targets , belong to the super family of integral membrane proteins characterized by seven transmembrane helices . Because they are difficult to crystallize , the three dimensional structure of these receptors have not yet been determined by X-ray crystallography , except one . In the absence of a 3-D structure , in-silico approaches for solving the structure of this class of proteins are widely used and provide valuable information for structure based drug design . There are several web servers and computer programs available that automate the modelling process of GPCRs . Some of these include Modeller , Swiss-Model server , Homer , etc . Using these tools reliable homology models of human histamine H1 receptor ( P35367 ) and thrombin receptor ( P25116 ) have been generated which explain the binding mode of the standard antagonists of these receptors and may be useful in designing their novel antagonists . Thrombin induces slug-mediated P12830 transcriptional repression and the parallel up-regulation of P19022 by a transcription-independent mechanism in Q96AT9 cells . The proliferation , directional migration to the vitreous and epithelial-mesenchymal transition ( EMT ) of quiescent , differentiated retinal pigment epithelium ( Q96AT9 ) cells is a major feature in the development of proliferative vitreoretinopathy ( P15151 ) following exposure of the immuno-privileged eye niche to serum components , thrombin among them . We have previously documented thrombin induction of Q96AT9 cell proliferation and migration . We here analyzed the effect of thrombin on the E/N cadherin switch , a hallmark of EMT . Results show that thrombin induces the specific repression of epithelial P12830 gene transcription , alongside with the up-regulation of mesenchymal P19022 protein in Q96AT9 cells . We demonstrate , for the first time , that thrombin induces P12830 repression by stimulating snail-2 ( O43623 ) transcription factor expression , and the concomitant up-regulation of P19022 through the transcription-independent increase in protein translation promoted by PI3K/PKC-ζ/ P42345 signaling . Our present findings suggest that the activation of protease-activated receptor-1 ( P25116 ) by thrombin induces EMT of Q96AT9 cells , further supporting a central role for thrombin in P15151 pathogenesis . High loading dose of clopidogrel is unable to satisfactorily inhibit platelet reactivity in patients with glycoprotein IIIA gene polymorphism : a genetic substudy of PRAGUE-8 trial . The study aimed to assess the impact of nine polymorphisms of genes encoding platelet receptors , enzymes , and hemostatic factors on clopidogrel efficacy to inhibit platelet reactivity in patients with stable coronary artery disease undergoing elective coronary angiography either with or without ad hoc percutaneous coronary intervention . The study was performed as a genetic substudy of the PRAGUE-8 trial . Ninety-five patients pretreated with 600 mg clopidogrel at least 6 h prior to coronary angiography were tested . Baseline platelet reactivity to ADP was assessed before the drug was administered . DB00758 efficacy was tested again at 12 and 28 h after administration . Polymorphisms of platelet receptors , glycoprotein ( GP ) Ia ( 807C/T ) , Q9HCN6 ( 13254C/T ) , P05106 ( PlA1/PlA2 ) , P25116 ( IVSn-14A/T ) , Q9H244 ( 32C/T ) , Q9H244 ( H1/H2 ) haplotype , gene variations of cyclooxygenase-1 , Leiden , and factor II mutations were studied . Flow cytometric tests of vasodilator-stimulated phosphoprotein phosphorylation states were used as a measure of drug efficacy . None of the gene polymorphisms influenced baseline ADP-induced platelet reactivity significantly . Twenty-eight hours after drug administration , differences in suppression of ADP-induced platelet reactivity were observed between polymorphism-positive and polymorphism-negative patients . Inhibition of platelet reactivity , after 600 mg of clopidogrel , was significantly less in carriers of PlA2 ( P=0.009 ) for mean decrease in platelet reactivity index . The proportion of clopidogrel nonresponders ( platelet reactivity index > 50 % ) was apparently higher in PlA2 carriers in comparison with PlA1/PlA1 patients ( 54 vs. 24 % , P=0.082 ) . A 600 mg loading dose of clopidogrel failed to acceptably inhibit platelet reactivity in patients who were positive for the PlA2 polymorphism . Presence of functionally active protease-activated receptors 1 and 2 in myenteric glia . Protease-activated receptors ( PARs ) belong to the family of membrane receptors coupled to G-proteins ; their presence is reported in a wide variety of cells . The object of this study was to demonstrate the presence of P25116 and P55085 in myenteric glia of the guinea pig , and to elucidate the cellular mechanisms that are triggered upon receptor activation . Thrombin and P25116 agonist peptide ( P09874 ) activate P25116 with a maximum mean +/- SEM change in intracellular calcium concentration with respect to basal level ( Delta[Ca2+]i ) of 183 +/- 18 nm and 169 +/- 6 nm , respectively . Trypsin and P55085 agonist peptide ( Q9UGN5 ) activate P55085 with a maximum Delta[Ca2+]i of 364 +/- 28 nm and 239 +/- 19 nm , respectively . Inhibition of phospholipase C by U73312 ( 1 microm ) decreased the Delta[Ca2+]i due to P25116 activation from 167 +/- 10 nm to 87 +/- 6 nm . The P55085 -mediated Delta[Ca2+]i decreased from 193 +/- 10 nm to 124 +/- 8 nm when phospholipase C activity was inhibited . Blockade of sphingosine kinase with dimethylsphingosine ( 1 microm ) decreased the Delta[Ca2+]i due to P55085 activation from 149 +/- 19 nm to 67 +/- 1 nm , but did not influence the P25116 -mediated Delta[Ca2+]i . P25116 and P55085 were localized in myenteric glia by immunolabeling . Our results indicate that P25116 and P55085 are present in myenteric glia of the guinea pig , and their activation leads to increases in intracellular calcium via different signal transduction mechanisms that involve activation of phospholipase C and sphingosine kinase . P62158 inhibitors induce spinal analgesia in rats . DB01373 is an important intracellular messenger that interacts with Ca(2+)-binding proteins , such as calmodulin ( P62158 ) , to activate several intracellular enzymes . The involvement of Ca2+ in the transmission of nociceptive signals has been demonstrated at the spina level . Specifically , spinal sensitization induced by persistent nociceptive stimulation seems to be related to an increase of cytosolic calcium and the subsequent activation of several enzymes , some of which are Ca2+/ P62158 dependent . In order to elucidate the possible implication of calmodulin in these pain processes , we have studied the effect of two calmodulin inhibitors ( W-7 and calmidazolium ) or the formalin and tail-flick tests in rats after their intrathecal administration . Antinociceptive effects were observed in both tests by injecting 0.12-1 mumol/rat of calmidazolium and 0.25-2 mumol/rat of W-7 . Calmidazolium was more potent than W-7 in inhibiting both phases of the formalin test , whereas lower doses of W-7 in comparison to calmidazolium affected the tail-flick latencies . In addition , both drugs induced , at high doses , a muscular flaccidity of the hindlimbs that impaired normal walking in the rats . This effect caused ; significant reduction of the rotarod performance when 1 mumol/rat of calmidazolium or 2 mumol/rat of W-7 were injected . Overall , our results show that calmodulin inhibitors are capable of producing spinal analgesia on phasic and tonic noxious stimuli in rats , thus rendering them a promising potential as analgesics . Characterization of the aggregation responses of camel platelets . BACKGROUND : Despite evidence of active hemostasis , camel platelets barely respond to common aggregating agents at standard doses used for human platelet aggregation . OBJECTIVES : The purpose of the study was to find out whether camel platelets can be activated by high doses or combinations of aggregation agonists , and to characterize the receptor that mediates the aggregation response to adenosine diphosphate ( ADP ) , the most potent agonist for camel platelets known so far . METHODS : Aggregation studies were performed with platelet-rich plasma ( PRP ) in response to multiple doses or combinations of ADP , epinephrine ( P08473 ) , collagen , and arachidonic acid ( AA ) . Aggregation responses to ADP were performed before and after the addition of the ADP receptor ( Q9H244 ) antagonist DB00758 . RESULTS : Camel platelets responded to ADP at doses higher than the standard dose for human platelets , and to combinations of P08473 and other agonists , while no aggregation was elicited with P08473 or AA alone . DB00758 blocked the ADP-induced aggregation responses in a dose-dependent fashion in vitro . CONCLUSIONS : Camel platelet aggregation can be activated by increasing the dose of some agonists such as ADP , but not AA or P08473 . Irreversible aggregation of camel platelets could also be triggered by a combination of P08473 and ADP , and collagen and AA . Inhibition with clopidogrel suggests that camel platelets express the ADP receptor , Q9H244 . Understanding platelet function in camels will add to the understanding of platelet function in health and disease . [ Anticoagulants of primary haemostasis ] . Inhibition of platelet function plays an important role in the treatment and secondary prevention of cardiovascular or cerebrovascular ischemic diseases . Established antiplatelet agents use different pharmacological targets for this role . Acetyl salicylic acid achieves a reduction of thromboxane A2 formation by inhibition of P23219 . DB00208 or clopidogrel are ADP- Q9H244 receptor antagonists . Tirofiban , abciximab or eptifibatid are used for the inhibition of the glycoprotein IIb/IIIa receptor which is activated at the surface of platelets preceding the final step of their aggregation . The mechanism of dipyridamole is based on the inhibition of adenosine uptake and of phosphodiesterase-5 . Efforts are made to improve antiplatelet therapy with the aim to find agents with favorable clinical outcome and lower bleeding risk . Current clinical studies focus on a new generation of ADP receptor antagonists ( prasugrel , cangrelor and ticagrelor ) as successors of ticlopidine and clopidogrel after coronary arterial interventions . Developments using platelet targets different from established drugs are thrombin receptor antagonists ( like SCH530348 ) or thromboxane receptor antagonists ( like S18886/terutroban ) in patients with cerebrovascular events . Results from recent experimental studies could lead to new strategies for antiplatelet therapy ( like inhibition of GP Ib receptor , GP VI receptor , platelet-leukocyte interaction , factor XII and others ) in the future . Store-operated Ca2+ entry ( SOCE ) induced by protease-activated receptor-1 mediates Q13586 protein phosphorylation to inhibit SOCE in endothelial cells through AMP-activated protein kinase and p38β mitogen-activated protein kinase . The Ca(2+) sensor Q13586 is crucial for activation of store-operated Ca(2+) entry ( SOCE ) through transient receptor potential canonical and Orai channels . Q13586 phosphorylation serves as an " off switch " for SOCE . However , the signaling pathway for Q13586 phosphorylation is unknown . Here , we show that SOCE activates AMP-activated protein kinase ( AMPK ) ; its effector p38β mitogen-activated protein kinase ( p38β MAPK ) phosphorylates Q13586 , thus inhibiting SOCE in human lung microvascular endothelial cells . Activation of AMPK using 5-aminoimidazole-4-carboxamide-1-β-d-ribofuranoside ( AICAR ) resulted in Q13586 phosphorylation on serine residues and prevented protease-activated receptor-1 ( P25116 ) -induced Ca(2+) entry . Furthermore , AICAR pretreatment blocked P25116 -induced increase in the permeability of mouse lung microvessels . Activation of SOCE with thrombin caused phosphorylation of isoform α1 but not α2 of the AMPK catalytic subunit . Moreover , knockdown of AMPKα1 augmented SOCE induced by thrombin . Interestingly , SB203580 , a selective inhibitor of p38 MAPK , blocked Q13586 phosphorylation and led to sustained Q13586 -puncta formation and Ca(2+) entry . Of the three p38 MAPK isoforms expressed in endothelial cells , p38β knockdown prevented P25116 -mediated Q13586 phosphorylation and potentiated SOCE . In addition , inhibition of the SOCE downstream target P62158 kinase kinase β ( CaMKKβ ) or knockdown of AMPKα1 suppressed P25116 -mediated phosphorylation of p38β and hence Q13586 . Thus , our findings demonstrate that SOCE activates CaMKKβ-AMPKα1-p38β MAPK signaling to phosphorylate Q13586 , thereby suppressing endothelial SOCE and permeability responses . An analysis of the effects of retinoic acid and other retinoids on the development of adrenergic cells from the avian neural crest . In the present work , we have investigated the role of all-trans-retinoic acid ( all-trans RA ) , and several other natural and synthetic retinoids , in the development of adrenergic cells in quail neural crest cultures . Dose response studies using all-trans RA and 13-cis RA revealed a dose-dependent increase in the number of adrenergic cells in neural crest cultures . Similar dose response studies using RA isomers and other natural retinoids did not result in the same increases . In order to determine the receptor mediating the effects of all-trans RA in the neural crest , we tested several synthetic analogs which specifically bind to a particular RA receptor ( RAR ) subtype . We found that the compound AM 580 , which activates the P10276 , produced an increase in adrenergic cells similar to that seen with all-trans RA . The compound DB02877 , which activates all RAR subtypes , also resulted in an increase in adrenergic cells . We conclude that the increase in adrenergic cells seen with all-trans RA is mediated by P10276 and possibly P10826 . To further define the actions of all-trans RA on the neural crest we incubated cultures with 5-bromo-2'-deoxyuridine ( BrdU ) to determine whether all-trans RA could affect the rate of proliferation . The results show that while all-trans RA did not increase the fraction of cells incorporating BrdU into their nuclei at early time points ( 24 h ) , it did increase BrdU incorporation by tyrosine hydroxylase ( TH ) positive cells at 5 days in culture . These findings demonstrate that the increase in adrenergic cells seen with all-trans RA in neural crest cultures is likely due to an increase in the proliferation of cells already expressing TH . Feasibility of a microarray-based point-of-care P33261 genotyping test for predicting clopidogrel on-treatment platelet reactivity . DB00758 is a prodrug which is converted into active metabolite by cytochrome P450 isoenzyme , P33261 . Numerous polymorphisms of P33261 are reported , and a strong link exists between loss-of-function ( LOF ) or gain-of-function polymorphisms , clopidogrel metabolism , and clinical outcome . Hence , a fully automated point-of-care P33261 genotyping assay is more likely to bring personalized antiplatelet therapy into real practice . We assessed the feasibility of the Verigene 2C19/ P35520 Nucleic Acid Test , a fully automated microarray-based assay , compared to bidirectional sequencing , and performed VerifyNow Q9H244 assay to evaluate the effect of P33261 polymorphisms on on-treatment platelet reactivity in 57 Korean patients treated with clopidogrel after percutaneous coronary intervention . The Verigene 2C19/ P35520 assay identified ∗2 , ∗3 , and ∗17 polymorphisms with 100 % concordance to bidirectional sequencing in 180 minutes with little hands-on time . Patients were classified into 4 groups : extensive ( ∗1/∗1 ; n = 12 , 21.1 % ) , intermediate ( ∗1/∗2 , ∗1/∗3 ; n = 33 , 57.9 % ) , poor ( ∗2/∗2 , ∗2/∗3 , and ∗3/∗3 ; n = 11 , 19.3 % ) , and ultrarapid metabolizers ( ∗1/∗17 ; n = 1 , 1.8 % ) . The prevalence of the CYP2C19 ∗2 , ∗3 , and ∗17 alleles was 36.0 % , 12.3 % , and 0.9 % . Platelet reactivity showed gene dose response according to the number of P33261 LOF allele . In conclusion , the Verigene 2C19/ P35520 assay gave accurate P33261 genotype results which were in well match with the differing on-treatment platelet reactivity . Targeting the PI3K/ P42345 axis , alone and in combination with autophagy blockade , for the treatment of malignant peripheral nerve sheath tumors . There is a critical need for efficacious therapeutic strategies to improve the outcome of patients afflicted by malignant peripheral nerve sheath tumors ( MPNST ) . Multiple lines of evidence suggest a role for deregulated phosphoinositide 3-kinase ( PI3K ) / P42345 signaling in MPNST , making this axis an attractive target for therapeutic manipulation . On the basis of previous observations obtained from in vitro experimentation , here we aimed to assess the effects of PI3K/ P42345 blockade on MPNST growth in vivo . The anti-MPNST impact of DB05241 , a dual PI3K/ P42345 inhibitor currently being evaluated in human cancer clinical trials , was tested in two human MPNST xenograft models ( STS26T and MPNST724 ) and an experimental model of pulmonary metastasis ( STS26T ) . DB05241 abrogated human MPNST local and metastatic growth in severe combined immunodeficient mice . Notably , this therapeutic approach failed to induce apoptosis in MPNST cells but rather resulted in marked productive autophagy . Importantly , genetic and pharmacologic autophagy blockade reversed apoptotic resistance and resulted in significant PI3K/ P42345 inhibition-induced MPNST cell death . The addition of the autophagy inhibitor , chloroquine , to the therapeutic regimen of MPNST xenografts after pretreatment with DB05241 resulted in superior antitumor effects as compared with either agent alone . Together , preclinical studies described here expand our previous findings and suggest that PI3K/ P42345 inhibition alone and ( most importantly ) in combination with autophagy blockade may comprise a novel and efficacious therapy for patients harboring MPNST . Characterization of antihistamines using biphasic cutaneous reaction in BALB/c mice . Effects of 11 histamine H1 receptor antagonists on IgE-mediated biphasic cutaneous reaction in mice were examined . The immediate phase reaction ( IPR ) assessed at 1 hour after antigen application was significantly inhibited by all antihistamines examined . The inhibition of IPR by cetirizine and mequitazine were potent , but those by cyproheptadine and diphenhydramine were weak . The later phase reaction ( LPR ) assessed at 24 hours after antigen application was inhibited by chlorpheniramine , oxatomide , ketotifen , mequitazine , emedastine , terfenadine and azelastine . The inhibition of LPR by emedastine was potent , but those by ketotifen and terfenadine were only partial . DB01084 inhibited both IPR and LPR comparably . Present results indicate that H1 receptor activation is involved in the IPR of the biphasic cutaneous reaction , and that the blockade of H1 receptors at IPR does not contribute to the attenuation of following LPR . P35367 antagonists inhibiting the LPR have a property distinct from H1 receptor antagonism , which may have an additional benefit for the treatment of allergic diseases . In vivo participation of nitric oxide in hyperproliferative epidermal phenomena in mice . A significant involvement of nitric oxide ( NO ) in the process of keratinocyte proliferation is reported with many divergences . To determine the involvement of NO in the hyperproliferative process of epidermis in vivo , non-selective inhibitor ( N(G)-nitro-L-arginine-methyl ester.HCl : L-NAME ) and selective inhibitors for inducible NO synthase ( P35228 ) and neuronal NO synthase ( P29475 ) ( DB02533 : AG and DB02207 : 7-NI , respectively ) and a NO-donor ( Sodium nitroprusside : SNP ) were topically applied twice a day in mice ear treated with multiple applications of croton oil . L-NAME and 7-NI treatments decreased and SNP increased ear edema formation . However , ear weight was reduced in groups that received L-NAME and 7-NI , while the AG and SNP groups presented an increment . The histological evaluation of epidermis thickness showed that all NOS inhibitors were able to prevent the increase in epidermis width caused by croton oil , while SNP contributed to enlargement . The same results were observed in the P12004 staining , where treatments with NOS inhibitors caused a reduction in the number of cells in the epidermis , while SNP caused an enhancement . 7-NI treatment reduced polymorphonuclear and mononuclear leukocytes migration when compared to the control group . The AG application increased the migration of polymorphonuclear and mononuclear cells , while the SNP enhanced only the polymorphonuclear cells . Therefore , in the skin NO produced by P29475 is involved in the control of keratinocyte hyperproliferation , with the contribution of P35228 . In the animal model of cutaneous chronic inflammation by croton oil , NO is involved in the exudation and leukocyte migration , with participation of all three enzymes . Effect of firocoxib or flunixin meglumine on recovery of ischemic-injured equine jejunum . OBJECTIVE : To determine whether treatment of horses with firocoxib affects recovery of ischemic-injured jejunum , while providing effective analgesia . ANIMALS : 18 horses . PROCEDURES : Horses ( n = 6 horses/group ) received saline ( 0.9 % NaCl ) solution ( 1 mL/50 kg , IV ) , flunixin meglumine ( 1.1 mg/kg , IV , q 12 h ) , or firocoxib ( 0.09 mg/kg , IV , q 24 h ) before 2 hours of jejunal ischemia . Horses were monitored via pain scores and received butorphanol for analgesia . After 18 hours , ischemic-injured and control mucosa were placed in Ussing chambers for measurement of transepithelial resistance and permeability to lipopolysaccharide . Histomorphometry was used to determine denuded villus surface area . Western blots for cyclooxygenase ( P36551 ) -1 and P35354 were performed . Plasma thromboxane B(2) and prostaglandin E(2) metabolite ( PGEM ) concentrations were determined . RESULTS : Pain scores did not significantly increase after surgery in horses receiving flunixin meglumine or firocoxib . Transepithelial resistance of ischemic-injured jejunum from horses treated with flunixin meglumine was significantly lower than in saline- or firocoxib-treated horses . Lipopolysaccharide permeability across ischemic-injured mucosa was significantly increased in horses treated with flunixin meglumine . Treatment did not affect epithelial restitution . P23219 was constitutively expressed and P35354 was upregulated after 2 hours of ischemia . Thromboxane B(2) concentration decreased with flunixin meglumine treatment but increased with firocoxib or saline treatment . Flunixin meglumine and firocoxib prevented an increase in PGEM concentration after surgery . CONCLUSIONS AND CLINICAL RELEVANCE : Flunixin meglumine retarded mucosal recovery in ischemic-injured jejunum , whereas firocoxib did not . Flunixin meglumine and firocoxib were effective visceral analgesics . DB09217 may be advantageous in horses recovering from ischemic intestinal injury . High-density genetic map of the P38398 region of chromosome 17q12-q21 . To facilitate the positional cloning of the breast-ovarian cancer gene P38398 , we constructed a high-density genetic map of the 8.3-cM interval between D17S250 and GIP on chromosome 17q12-q21 . Markers were mapped by linkage in the CEPH and in extended kindreds in our breast cancer series . The map comprises 33 ordered polymorphisms , including 12 genes and 21 anonymous markers , yielding an average of one polymorphism every 250 kb . Twenty-five of the markers are PCR-based systems . The order of polymorphic genes and markers is cen-D17S250-D17S518- P04626 - P10827 - P10276 -D17S80 - P13645 -[D17S800-D17S857]-GAS- D17S856-EDH17B-D17S855-D17S859-D17S858- [ ++ + P01298 -D17S78 ] -D17S183- P02730 -D17S579- D17S509- [ D17S508-D17S190 = D17S810 ] -D17S791- [ D17S181 = D17S806 ] -D17S797- P17509 - P05106 - [ D17S507 = GIP ] -qter . P38398 lies in the middle of the interval , between P10827 and D17S183 . Markers from this map can be used to determine whether cancer is linked to P38398 in families , to evaluate whether tumors have lost heterozygosity at loci in the region , and to identify probes for characterizing chromosomal rearrangements from patients and from tumors .

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interacts_with DB00317?

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Effects of ethanol and anesthetics on type 1 and 5 metabotropic glutamate receptors expressed in Xenopus laevis oocytes . Previous studies have demonstrated that ethanol and volatile anesthetics inhibit the function of some metabotropic ( G protein-coupled ) receptors , including the 5-hydroxytryptamine2 and muscarinic cholinergic receptors . The metabotropic glutamate receptors ( mGluRs ) show little sequence homology with most other metabotropic receptors and are important modulators of synaptic transmission in the mammalian central nervous system . It was of interest to determine drug actions on these receptors , and we investigated the effects of ethanol , halothane , the anesthetic compound P13726 ( 1-chloro-1,2,2-trifluorocyclobutane ) , and the nonanesthetics F6 ( 1,2-dichlorohexafluorocyclobutane ) and P00451 ( 2,3-chlorooctafluorobutane ) on the function of Q13255 and P41594 expressed in Xenopus laevis oocytes . Halothane , P13726 , and ethanol inhibited P41594 -induced Ca(2+)-dependent Cl- currents , yet pharmacologically relevant concentrations of these compounds had little effect on the glutamate-induced currents in the oocytes expressing Q13255 . F6 had inhibitory effects on both receptors , and P00451 did not affect either Q13255 or P41594 function . The protein kinase C ( PKC ) inhibitor GF109203X enhanced the glutamate-induced current , and the PKC activator phorbol-12-myristate-13-acetate inhibited this current in the oocytes expressing P41594 , but these compounds had little effect on Q13255 function . GF109203X abolished the inhibitory effects of halothane , P13726 , and ethanol on mGluR5s . Conversely , the phosphatase inhibitor calyculin A prolonged the action of halothane and ethanol . Furthermore , mutation of a PKC consensus site ( Ser890 ) of P41594 abolished the inhibitory effects of halothane , P13726 , and ethanol . These results suggest that ethanol and volatile anesthetics inhibit P41594 because they promote PKC-mediated phosphorylation . P05231 sensitizes prostate cancer to the antiproliferative effect of IFNα2 through Q00978 . Development and progression of prostate cancer ( PCa ) are associated with chronic inflammation . The cytokine interleukin 6 ( P05231 ) can influence progression , differentiation , survival , and angiogenesis of PCa . To identify novel pathways that are triggered by P05231 , we performed a gene expression profiling of two PCa cell lines , LNCaP and MDA PCa 2b , treated with 5 ng/ml P05231 . Interferon ( IFN ) regulatory factor 9 ( Q00978 ) was identified as one of the most prevalent P05231 -regulated genes in both cell lines . Q00978 is a mediator of type I IFN signaling and acts together with P42224 and 2 to activate transcription of IFN-responsive genes . The P05231 regulation of Q00978 was confirmed at mRNA and protein levels by quantitative real-time PCR and western blot respectively in both cell lines and could be blocked by the anti- P05231 antibody DB09036 . Three PCa cell lines , PC3 , Du-145 , and LNCaP- P05231 + , with an autocrine P05231 loop displayed high expression of Q00978 . A tissue microarray with 36 PCa tissues showed that Q00978 protein expression is moderately elevated in malignant areas and positively correlates with the tissue expression of P05231 . Downregulation and overexpression of Q00978 provided evidence for an IFN-independent role of Q00978 in cellular proliferation of different PCa cell lines . Furthermore , expression of Q00978 was essential to mediate the antiproliferative effects of IFNα2 . We concluded that P05231 is an inducer of Q00978 expression in PCa and a sensitizer for the antiproliferative effects of IFNα2 . [ New molecular targeting drugs for metastatic colorectal cancer ] . I explain an outline of the molecular target of new drugs for colorectal cancer . First , regorafenib is an orally active , multikinase inhibitor . Second , as angiogenesis inhibitors , aflibercept and ramucirumab . DB08885 is the second anti-angiogenic agent after bevacizumab with a FDA approval for metastatic colorectal cancer . DB05578 is a fully human mAb that binds human P35968 , thus blocking P15692 binding and inhibiting angiogenesis . Finally , I introduce a new combination chemotherapy regimen with molecular targetting drugs . High expression of growth factors and growth factor receptors in ovarian metastases from ileal carcinoids : an immunohistochemical study of 2 cases . OBJECTIVE AND DESIGN : Ovarian metastatic carcinoids are rare neoplasms that show prominent fibrosis of tumor stroma and are often associated with peritoneal carcinomatosis . We studied formalin-fixed and paraffin-embedded tumor specimens of 2 cases of ovarian metastases from ileal enterochromaffin cell carcinoids immunohistochemically to evaluate whether acidic fibroblast growth factor ( P05230 ) , transforming growth factor-alpha ( TGFalpha ) , and their respective receptors ( fibroblast growth factor receptor-4 [ P22455 ] and epidermal growth factor receptor [ P00533 ] ) may play a role in the pathogenesis of stromal fibroblast reaction and in the mechanism of tumor dissemination . RESULTS : In both cases , the majority of tumor cells expressed immunoreactivity for P05230 , P22455 , and TGFalpha . Immunoreactivity for P22455 was detected in stromal cells of both cases , while P00533 -positive stromal cells were found in only 1 case . Immunoreactivity for P22455 was also found in peritoneal mesothelial cells . CONCLUSIONS : The coexpression of P05230 and P22455 in neoplastic enterochromaffin cells suggests that P05230 may act as an autocrine factor stimulating tumor cell growth . In addition , P05230 and TGFalpha may stimulate , in a paracrine fashion , the proliferation of P22455 - and P00533 -immunoreactive stromal fibroblasts . Finally , interaction of P05230 -immunoreactive enterochromaffin cells with P22455 -bearing mesothelial cells may play a role in the mechanism of serosal implant and spread of tumor cells . P00533 variant III mutations in lung tumorigenesis and sensitivity to tyrosine kinase inhibitors . The tyrosine kinase inhibitors gefitinib ( DB00317 ) and erlotinib ( Tarceva ) have shown anti-tumor activity in the treatment of non-small cell lung cancer ( NSCLC ) . Dramatic and durable responses have occurred in NSCLC tumors with mutations in the tyrosine kinase domain of the epidermal growth factor receptor ( P00533 ) . In contrast , these inhibitors have shown limited efficacy in glioblastoma , where a distinct P00533 mutation , the variant III ( vIII ) in-frame deletion of exons 2-7 , is commonly found . In this study , we determined that EGFRvIII mutation was present in 5 % ( 3/56 ) of analyzed human lung squamous cell carcinoma ( SCC ) but was not present in human lung adenocarcinoma ( 0/123 ) . We analyzed the role of the EGFRvIII mutation in lung tumorigenesis and its response to tyrosine kinase inhibition . Tissue-specific expression of EGFRvIII in the murine lung led to the development of NSCLC . Most importantly , these lung tumors depend on EGFRvIII expression for maintenance . Treatment with an irreversible P00533 inhibitor , HKI-272 , dramatically reduced the size of these EGFRvIII-driven murine tumors in 1 week . Similarly , Ba/ P13726 cells transformed with the EGFRvIII mutant were relatively resistant to gefitinib and erlotinib in vitro but proved sensitive to HKI-272 . These findings suggest a therapeutic strategy for cancers harboring the EGFRvIII mutation . Characterization of the interaction of ingenol 3-angelate with protein kinase C . DB05013 ( I3A ) is one of the active ingredients in Euphorbia peplus , which has been used in traditional medicine . Here , we report the initial characterization of I3A as a protein kinase C ( PKC ) ligand . I3A bound to P17252 in the presence of phosphatidylserine with high affinity ; however , under these assay conditions , little PKC isoform selectivity was observed . PKC isoforms did show different sensitivity and selectivity for down-regulation by I3A and phorbol 12-myristate 13-acetate ( PMA ) in WEHI-231 , Q9BPY8 -92 , and Colo-205 cells . In all of the three cell types , I3A inhibited cell proliferation with somewhat lower potency than did PMA . In intact CHO- P04264 cells , I3A was able to translocate different green fluorescent protein-tagged PKC isoforms , visualized by confocal microscopy , with equal or higher potency than PMA . PKC-delta in particular showed a different pattern of translocation in response to I3A and PMA . I3A induced a higher level of secretion of the inflammatory cytokine interleukin 6 compared with PMA in the WEHI-231 cells and displayed a marked biphasic dose-response curve for the induction . I3A was unable to cause the same extent of association of the C1b domain of PKC-delta with lipids , compared with PMA or the physiological regulator diacylglycerol , and was able to partially block the association induced by these agents , measured by surface plasmon resonance . The in vitro kinase activity of P17252 induced by I3A was lower than that induced by PMA . The novel pattern of behavior of I3A makes it of great interest for further evaluation . Molecular correlates of gefitinib responsiveness in human bladder cancer cells . We characterized the effects of the small molecule epidermal growth factor receptor ( P00533 ) inhibitor gefitinib ( ZD1839 , DB00317 ) on cell proliferation in a panel of 17 human bladder cancer cell lines . Gefitinib inhibited DNA synthesis in a concentration-dependent fashion in 6 of 17 lines . Growth inhibition was associated with p27(Kip1) accumulation and decreased cyclin-dependent kinase 2 activity . Gefitinib also inhibited baseline P00533 , AKT , and extracellular signal-regulated kinase ( P29323 ) phosphorylation in the P00533 -dependent cells maintained in serum-free medium , whereas it had no effect on baseline P00533 or P29323 phosphorylation in the P00533 -independent cells . Analyses of candidate markers of P00533 dependency revealed that the gefitinib-sensitive cells expressed higher surface P00533 levels than the gefitinib-resistant lines . Gefitinib-sensitive cells generally expressed higher levels of P12830 and lower levels of vimentin than the gefitinib-resistant cells , but these correlations were not perfect , suggesting that these markers of epithelial-mesenchymal transition can not be used by themselves to prospectively predict P00533 -dependent growth . Together , our results show that bladder cancer cells are markedly heterogeneous with respect to their sensitivity to P00533 antagonists . Although surface P00533 levels and epithelial-mesenchymal transition status seem to roughly correlate with responsiveness , they can not be used by themselves to identify bladder tumors that will be sensitive to P00533 -directed therapy . However , comparing levels of p27(Kip1) or DNA synthesis before and after gefitinib exposure does identify the drug-sensitive cells . Coadministrating luteolin minimizes the side effects of the aromatase inhibitor letrozole . P11511 inhibitors ( AIs ) have been used as adjuvant therapeutic agents for breast cancer . Their adverse side effect on blood lipid is well documented . Some natural compounds have been shown to be potential AIs . In the present study , we compared the efficacy of the flavonoid luteolin to the clinically approved AI letrozole ( DB01006 ; Novartis Pharmaceuticals , East Hanover , NJ ) in a cell and a mouse model . In the in vitro experimental results for aromatase inhibition , the Ki values of luteolin and letrozole were estimated to be 2.44 µM and 0.41 nM , respectively . Both letrozole and luteolin appeared to be competitive inhibitors . Subsequently , an animal model was used for the comparison . P11511 -expressing MCF-7 cells were transplanted into ovariectomized athymic mice . Luteolin was given by mouth at 5 , 20 , and 50 mg/kg , whereas letrozole was administered by intravenous injection . Similar to letrozole , luteolin administration reduced plasma estrogen concentrations and suppressed the xenograft proliferation . The regulation of cell cycle and apoptotic proteins-such as a decrease in the expression of Bcl-xL , cyclin-A/D1/E , P24941 /4 , and increase in that of Bax-was about the same in both treatments . The most significant disparity was on blood lipids . In contrast to letrozole , luteolin increased fasting plasma high-density lipoprotein concentrations and produced a desirable blood lipid profile . These results suggested that the flavonoid could be a coadjuvant therapeutic agent without impairing the action of AIs . Moving beyond chemotherapy : novel cytostatic agents for malignant mesothelioma . It is now known that vascular endothelial growth factor ( P15692 ) and platelet derived growth factor ( PDGF ) are autocrine growth factors in malignant mesothelioma ; epidermal growth factor receptor ( P00533 ) is also highly overexpressed . Cytotoxic drugs that target these growth factors offer fresh potential for the treatment of mesothelioma . Clinical trials have recently been initiated to evaluate the anti-tumour activity of the P15692 inhibitors SU5416 , bevacizumab and thalidomide . ZD1839 ( DB00317 , AstraZeneca ) , an inhibitor of P00533 tyrosine kinase , is also being evaluated . Two clinical trials are planned to evaluate the two PDGF inhibitors Gleevec ( Imatinib mesylate , STI-571 , Novartis Pharmaceuticals ) and PTK787 ( Novartis Pharmaceuticals ) . Critical role of both p27KIP1 and p21CIP1/ P38936 in the antiproliferative effect of ZD1839 ( ' DB00317 ' ) , an epidermal growth factor receptor tyrosine kinase inhibitor , in head and neck squamous carcinoma cells . High expression of the epidermal growth factor receptor ( P00533 ) has been implicated in the development of squamous-cell carcinomas of head and neck ( SCCHN ) . ZD1839 ( ' DB00317 ' ) is an orally active , selective P00533 -TKI ( P00533 -tyrosine kinase inhibitor ) that blocks signal transduction pathways implicated in proliferation and survival of cancer cells , and other host-dependent processes promoting cancer growth . We have demonstrated that ZD1839 induces growth arrest in SCCHN cell lines by inhibiting P00533 -mediated signaling . Cell cycle kinetic analysis demonstrated that ZD1839 induces a delay in cell cycle progression and a P55008 arrest together with a partial G2/M block ; this was associated with increased expression of both p27( P46527 ) and P38936 (CIP1/ P38936 ) cyclin-dependent kinase ( CDK ) inhibitors . The activity of P24941 , the main target of CIP/ Q99828 CDK inhibitors , was reduced in a dose-dependent fashion after 24 h of ZD1839 treatment and this effect correlated to the increased amount of p27( P46527 ) and P38936 (CIP1/ P38936 ) proteins associated with P24941 -cyclin-E and P24941 -cyclin-A complexes . In addition , ZD1839-induced growth inhibition was significantly reduced in cell transfectants expressing p27( P46527 ) or P38936 (CIP1/ P38936 ) antisense constructs . Overall , these results as well as the timing of the effect of ZD1839 on P55008 arrest and p27( P46527 ) and P38936 (CIP1/ P38936 ) upregulation , suggest a mechanistic connection between these events . P00533 ( P00533 ) transactivation by estrogen via the G-protein-coupled receptor , Q99527 : a novel signaling pathway with potential significance for breast cancer . The biological and biochemical effects of estrogen have been ascribed to its known receptors , which function as ligand-inducible transcription factors . However , estrogen also triggers rapid activation of classical second messengers ( DB02527 , calcium , and inositol triphosphate ) and stimulation of intracellular signaling cascades mitogen-activated protein kinase ( Q96HU1 K ) , PI3K and P29474 . These latter events are commonly activated by membrane receptors that either possess intrinsic tyrosine kinase activity or couple to heterotrimeric G-proteins . We have shown that estrogen transactivates the epidermal growth factor receptor ( P00533 ) to Q96HU1 K signaling axis via the G-protein-coupled receptor ( GPCR ) , Q99527 , through the release of surface-bound proHB- P01133 from estrogen receptor ( ER ) -negative human breast cancer cells [ Molecular Endocrinology 14 ( 2000 ) 1649 ] . This finding is consistent with a growing body of evidence suggesting that transactivation of EGFRs by GPCRs is a recurrent theme in cell signaling . GPCR-mediated transactivation of EGFRs by estrogen provides a previously unappreciated mechanism of cross-talk between estrogen and serum growth factors , and explains prior data reporting the P01133 -like effects of estrogen . This novel mechanism by which estrogen activates growth factor-dependent signaling and its implications for breast cancer biology are discussed further in this review . The influence of ionotropic and metabotropic glutamate receptor ligands on anxiety-like effect of amphetamine withdrawal in rats . Chronic amphetamine use results in anxiety-like states after drug cessation . The aim of the study was to determine a role of ionotropic and metabotropic glutamate receptor ligands in amphetamine-evoked withdrawal anxiety in the elevated plus-maze test in rats . In our study memantine ( 8 and 12 mg/kg ) , a noncompetitive N-methyl-d-aspartate ( DB01221 ) receptor antagonist did not reduce amphetamine withdrawal anxiety . DB00659 ( DB01221 and metabotropic glutamate 5 receptor ( P41594 ) antagonist ) at the dose 200 and 400mg/kg showed anxiolytic-like effect , thus increasing the percent of time spent in open arms and a number of open arm entries . P41594 selective antagonist , MTEP ( 3-[(2-methyl-1,3-thiazol-4-yl)ethynyl]pyridine hydrochloride ) and Q14416 /3 agonist , LY354740 ( 1S,2S,5R,6S ) -2-aminobicyclo[3.1.0]hexane-2,6-dicarboxylic acid ) , caused effects similar to acamprosate at doses 1.25-5mg/kg and 2.5-5mg/kg , respectively . None of the glutamate ligands influenced locomotor activity of rats when given to the saline-treated group . Taking into account the positive correlation between amphetamine withdrawal-induced anxiety and relapse to amphetamine taking , our results suggest that modulation of mGluRs may prevent relapse to amphetamine and might pose a new direction in amphetamine abuse therapy . P15121 regulates high glucose-induced ectodomain shedding of tumor necrosis factor ( P01375 ) -alpha via protein kinase C-delta and P01375 converting enzyme in vascular smooth muscle cells . Chronic low-grade inflammation has emerged as a key contributor to the cardiovascular complications of diabetes , however , the mechanisms by which diabetes increases inflammation remain poorly understood . Here , we report that exposure to high glucose ( HG ) stimulates ectodomain shedding of P01375 from rat aortic smooth muscle cells in culture . Our results show that exposure to HG decreases membrane-associated P01375 . This decrease in unprocessed P01375 was prevented by the aldose reductase ( AR ) inhibitor sorbinil and AR small interference RNA . Treatment with HG , but not equimolar mannitol or 3-O-methyl glucose , resulted in phosphorylation and activation of P01375 converting enzyme ( P78536 ) ( P78536 ) , which were attenuated by sorbinil or AR-specific small interference RNA . HG-induced P78536 phosphorylation and P01375 processing were also prevented by P01375 protease inhibitor-1 , an inhibitor of P78536 . Inhibition of protein kinase C ( PKC ) -delta by rottlerin prevented HG-induced P78536 activation and the accumulation of unprocessed P01375 . Treatment with sorbinil decreased elevated levels of circulating P01375 in streptozotocin-treated diabetic rats . DB02712 treatment also decreased the expression of P01375 , matrix metalloproteinase-2 , matrix metalloproteinase-9 , and increased tissue inhibitor of metalloproteinase-3 in vascular smooth muscle cells treated with HG and in balloon-injured carotid arteries of diabetic rats . These results indicate that HG-induced P01375 shedding could be attributed to P78536 activation , which is regulated , in part , by PKC-delta and AR . Therefore , inhibition of P78536 by P01375 protease inhibitor-1 , or pharmacological inhibition of PKC-delta or AR may represent useful strategies for treating vascular inflammation associated with diabetes . Dimerization effect of sucrose octasulfate on rat P05230 . Fibroblast growth factors ( FGFs ) constitute a family of at least 23 structurally related heparin-binding proteins that are involved in regulation of cell growth , survival , differentiation and migration . DB01901 ( SOS ) , a chemical analogue of heparin , has been demonstrated to activate FGF signalling pathways . The structure of rat P05230 crystallized in the presence of SOS has been determined at 2.2 A resolution . SOS-mediated dimerization of P05230 was observed , which was further supported by gel-filtration experiments . The major contributors to the sulfate-binding sites in rat P05230 are Lys113 , Lys118 , Arg122 and Lys128 . An arginine at position 116 is a consensus residue in mammalian FGF molecules ; however , it is a serine in rat P05230 . This difference may be important for SOS-mediated P05230 dimerization in rat . Antagonism of Q9GZP0 by human antibody DB05139 prevents renal scarring in experimental glomerulonephritis . Glomerular mesangial cell proliferation and/or matrix accumulation characterizes many progressive renal diseases . Q9GZP0 was identified recently as a novel mediator of mesangial cell proliferation in vitro and in vivo . This study investigated the long-term consequences of Q9GZP0 inhibition in vivo . Rats with progressive mesangioproliferative glomerulonephritis ( uninephrectomy plus anti-Thy-1.1 antibody ) received the Q9GZP0 -neutralizing , fully human mAb DB05139 on days 3 , 10 , and 17 after disease induction . Glomerular mesangioproliferative changes on day 10 were significantly reduced by anti- Q9GZP0 treatment as compared with control antibody . Eight weeks after disease induction , anti- Q9GZP0 therapy significantly ameliorated focal segmental glomerulosclerosis , podocyte damage ( de novo desmin expression ) , tubulointerstitial damage , and fibrosis as well as the accumulation of renal interstitial matrix including type III collagen and fibronectin . Treatment with anti- Q9GZP0 also reduced the cortical infiltration of monocytes/macrophages on day 56 , possibly related to lower renal cortical complement activation ( C5b-9 deposition ) and/or reduced epithelial-to-mesenchymal transition ( preserved cortical expression of P12830 and reduced expression of vimentin and alpha-smooth muscle actin ) . In conclusion , these data provide evidence for a causal role of Q9GZP0 in the pathogenesis of renal scarring and point to a new therapeutic approach to progressive mesangioproliferative renal disease . Reduced neuronal injury after treatment with NG-nitro-L-arginine methyl ester ( L-NAME ) or 2-sulfo-phenyl-N-tert-butyl nitrone ( S-PBN ) following experimental brain contusion . OBJECTIVE : DB00435 ( NO ) and oxygen free radicals are implicated in the pathophysiology of traumatic brain injury ( TBI ) . Peroxynitrite formation from NO and superoxide contributes to secondary neuronal injury but the neuroprotective effects of nitric oxide synthase ( NOS ) -inhibitors have been contradictory . This study was undertaken to examine whether PTtic administration of the ( NOS ) -inhibitor DB04223 methyl ester ( L-NAME ) , and a combination of L-NAME and the nitrone radical scavenger 2-sulfo-phenyl-N-tert-butyl nitrone ( S-PBN ) favorable affects neuronal injury in a model of TBI . METHODS : A weight-drop model of TBI was used . The animals received L-NAME , S-PBN or a combination of the drugs 15 minutes prothrombin time ( PT ) and sacrificed after 24 hours or six days . NOS activity was measured by the conversion of L-[U-C]arginine to L-[U-C]citrulline . Peroxynitrite formation , cellular apoptosis , neuronal degeneration and survival were assessed by nitrotyrosine- , TUNEL- , Fluoro-Jade- and NeuN-stainings . RESULTS : P29474 and P29475 activity was significantly reduced in animals that received L-NAME alone or the combination with S-PBN. P35228 activity or P35228 immunoreactivity was not affected . All treatments significantly reduced neuronal degeneration and nitrotyrosine immunoreactivity at 24 hours and increased neuronal survival at six days PT . No differences were detected between L-NAME and L-NAME + S-PBN groups . CONCLUSION : NO from NOS contributes to secondary neuronal injury in this TBI-model . PTtic treatment does not inhibit early beneficial NO-related effects . L-NAME and S-PBN limit peroxynitrite formation , promoting neuronal survival . The combination of L-NAME and S-PBN was neuroprotective ; surprisingly no additive effects were found on nitrotyrosine formation , apoptosis or neuronal survival . Simultaneous inhibition of epidermal growth factor receptor ( P00533 ) signaling and enhanced activation of tumor necrosis factor-related apoptosis-inducing ligand ( P50591 ) receptor-mediated apoptosis induction by an scFv:sTRAIL fusion protein with specificity for human P00533 . P00533 ( P00533 ) signaling inhibition by monoclonal antibodies and P00533 -specific tyrosine kinase inhibitors has shown clinical efficacy in cancer by restoring susceptibility of tumor cells to therapeutic apoptosis induction . P01375 -related apoptosis-inducing ligand ( P50591 ) is a promising anti-cancer agent with tumor-selective apoptotic activity . Here we present a novel approach that combines P00533 -signaling inhibition with target cell-restricted apoptosis induction using a P50591 fusion protein with engineered specificity for P00533 . This fusion protein , scFv425:sTRAIL , comprises the P00533 -blocking antibody fragment scFv425 genetically fused to soluble P50591 ( sTRAIL ) . Treatment with scFv425:sTRAIL resulted in the specific accretion to the cell surface of P00533 -positive cells only . P00533 -specific binding rapidly induced a dephosphorylation of P00533 and down-stream mitogenic signaling , which was accompanied by cFLIP(L) down-regulation and Bad dephosphorylation . P00533 -specific binding converted soluble scFv425:sTRAIL into a membrane-bound form of P50591 that cross-linked agonistic P50591 receptors in a paracrine manner , resulting in potent apoptosis induction in a series of P00533 -positive tumor cell lines . Co-treatment of P00533 -positive tumor cells with the P00533 -tyrosine kinase inhibitor DB00317 resulted in a potent synergistic pro-apoptotic effect , caused by the specific down-regulation of O15519 . Furthermore , in mixed culture experiments binding (L)of scFv425:sTRAIL to P00533 -positive target cells conveyed a potent apoptotic effect toward P00533 -negative bystander tumor cells . The favorable characteristics of scFv425:sTRAIL , alone and in combination with DB00317 , as well as its potent anti-tumor bystander activity indicate its potential value for treatment of P00533 -expressing cancers . DB00317 strengthens the cytotoxic effect of docetaxel in NSCLC models that harbor specific molecular characteristics . Non-small cell lung cancer ( NSCLC ) is the most lethal malignant tumor and is also considered one of the most chemoresistant cancers . Despite the benefits obtained from platinum-based therapy , the majority of patients treated will progress and die . In the continuing quest for personalized therapy based on the biomolecular characteristics of each single patient , clinical practice now seems to be oriented towards combining conventional drugs with molecular-targeted agents . In the present study , we evaluated the antitumor activity of docetaxel , one of the most widely used drugs for second-line treatment , and DB00317 , an P00533 -targeting tyrosine kinase inhibitor , administered singly or in sequence . The study was performed on three human NSCLC cell lines ( ChaGo- P04264 , CAEP and RAL ) that exhibit different expression of proliferation and apoptosis-related markers , and do not harbor P00533 mutations . The efficacy of docetaxel and DB00317 differed in the three cell lines and an important synergistic interaction was observed with the sequence 1-h docetaxel --> 72-h DB00317 during which DB00317 doubled the fraction of docetaxel-induced apoptotic cells , amplifying a caspase-dependent apoptosis and inhibiting docetaxel-induced P38936 hyperexpression . Moreover , the important role of MAPK-dependent modulation of this molecular marker was shown using a specific inhibitor . The results from the present preclinical study demonstrate the cytotoxic activity of DB00317 and its ability to increase taxane activity in a model that does not harbor P00533 -specific mutations , thus highlighting the importance of focusing on alternative molecular targets of DB00317 activity . Significance of interleukin-6 signaling in the resistance of pharyngeal cancer to irradiation and the epidermal growth factor receptor inhibitor . PURPOSE : Tumor eradication by chemoradiotherapy for pharyngeal cancer has not been particularly successful . Targeting epithelial growth factor receptor ( P00533 ) could be a potential treatment strategy providing additional benefits , but only a subset of these tumors gives a clinically significant response to P00533 inhibitors . The aim has been to identify the role of interleukin-6 ( P05231 ) signaling and its predictive power in the treatment response of pharyngeal cancer . METHODS AND MATERIALS : Human pharyngeal cancer cell lines , including the hypopharyngeal cancer cell line FaDu and its derived cell line FaDu-C225-R , were selected . Changes in tumor growth , response to treatment , and responsible signaling pathway were investigated in vitro . Furthermore , 95 pharyngeal cancer tissue specimens were analyzed by immunohistochemical staining , and correlations were made between levels of P05231 , P05231 receptor ( IL-6R ) , p-AKT , and p- P40763 expression and the clinical outcome of patients . RESULTS : In vitro , either extrinsic P05231 stimulation of cancer cells or intrinsically activated P05231 signaling detected in FADu-C225-R cells results in resistance to irradiation and P00533 inhibitor . Blocking P05231 signaling attenuated aggressive tumor behavior and sensitized the cells to treatments . The responsible mechanisms included decreased p- P40763 , less nuclear translocation of P00533 , and subsequently attenuated epithelial-mesenchymal transition . Regarding clinical data , staining of p- P40763 and P05231 was significantly linked with lower response rates to treatments and shorter survival in pharyngeal cancer patients . CONCLUSIONS : P05231 and p- P40763 may be significant predictors of pharyngeal carcinoma , and regulating P05231 signaling can be considered a promising therapeutic approach . Differential radiosensitisation by ZD1839 ( DB00317 ) , a highly selective epidermal growth factor receptor tyrosine kinase inhibitor in two related bladder cancer cell lines . The epidermal growth factor receptor ( P00533 ) is expressed in a wide variety of epithelial tumours including carcinoma of the bladder . Stimulation of the P00533 pathway is blocked by ZD1839 ( DB00317 ) , a highly selective P00533 tyrosine kinase inhibitor . Radical radiotherapy is an established organ sparing treatment option for muscle invasive bladder cancer and this study has explored the possibility for the use of ZD1839 as a radiosensitiser in this scenario . The effect of combination treatment with ZD1839 ( 0.01 microM ) and ionising radiation in the established bladder cancer cell lines MGH-U1 and its radiosensitive mutant clone S40b was measured by clonogenic assays . A highly significant radiosensitising effect was seen in both cell lines ( P < 0.001 for MGH-U1 and S40b cell lines ) . This effect was independent of the concentration of the drug and the duration of exposure prior to treatment with ionising radiation . Cell cycle kinetics of both cell lines was not significantly altered with ZD1839 ( 0.01 microM ) as a single agent . A modest induction of apoptosis was observed with ZD1839 ( 0.01 microM ) as a single agent , but a marked induction was observed with the combination treatment of ZD1839 and ionising radiation . These results suggest a potentially important role for ZD1839 in combination with radiotherapy in the treatment of muscle invasive bladder cancer .

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interacts_with DB00588?

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Q9H3N8 antagonism diminishes existing airway inflammation and dysfunction via modulation of Th2 cytokines . BACKGROUND : Airway remodeling and dysfunction are characteristic features of asthma thought to be caused by aberrant production of Th2 cytokines . Q9H3N8 ( Q9H3N8 ) perturbation has previously been shown to modify acute inflammation and Th2 cytokine production in a murine model of asthma . We examined the ability of Q9H3N8 antagonists to therapeutically modify the effects of Th2 cytokine production such as goblet cell hyperplasia ( P30793 ) , and collagen deposition in a sub-chronic model of asthma . In addition , effects on Th2 mediated lung dysfunction were also determined . METHODS : Mice were sensitized to ovalbumin ( OVA ) followed by repeated airway challenge with OVA . After inflammation was established mice were dosed with the Q9H3N8 antagonist , JNJ 7777120 , or anti- P35225 antibody for comparison . Airway hyperreactivity ( P35869 ) was measured , lungs lavaged and tissues collected for analysis . RESULTS : Therapeutic Q9H3N8 antagonism inhibited T cell infiltration in to the lung and decreased Th2 cytokines P35225 and P05113 . P35225 dependent remodeling parameters such as P30793 and lung collagen were reduced . Intervention with Q9H3N8 antagonist also improved measures of central and peripheral airway dysfunction . CONCLUSIONS : These data demonstrate that therapeutic Q9H3N8 antagonism can significantly ameliorate allergen induced , Th2 cytokine driven pathologies such as lung remodeling and airway dysfunction . The ability of Q9H3N8 antagonists to affect these key manifestations of asthma suggests their potential as novel human therapeutics . DB00472 and all other SSRIs are P41595 Agonists - Importance for their Therapeutic Effects . DB00472 and other serotonin-specific re-uptake inhibitors ( SSRIs ) are generally thought to owe their therapeutic potency to inhibition of the serotonin transporter ( P31645 ) . However , research in our laboratory showed that it affects , with relatively high affinity the P41595 receptor in cultured astrocytes ; this finding was confirmed by independent observations showing that fluoxetine loses its ability to elicit SSRI-like responses in behavioral assays in mice in which the P41595 receptor was knocked-out genetically or inhibited pharmacologically . All clinically used SSRIs are approximately equipotent towards P41595 receptors and exert their effect on cultured astrocytes at concentrations similar to those used clinically , a substantial difference from their effect on P31645 . We have demonstrated up-regulation and editing of astrocytic genes for ADAR2 , the kainate receptor Q13002 , P47712 and the P41595 receptor itself after chronic treatment of cultures , which do not express P31645 and after treatment of mice ( expressing P31645 ) for 2 weeks with fluoxetine , followed by isolation of astrocytic and neuronal cell fractionation . Affected genes were identical in both experimental paradigms . DB00472 treatment also altered Ca(2+) homeostatic cascades , in a specific way that differs from that seen after treatment with the anti-bipolar drugs carbamazepine , lithium, or valproic acid . All changes occurred after a lag period similar to what is seen for fluoxetine 's clinical effects , and some of the genes were altered in the opposite direction by mild chronic inescapable stress , known to cause anhedonia , a component of major depression . In the anhedonic mice these changes were reversed by treatment with SSRIs . DB00092 promotes co-stimulation blockade based allograft survival in nonhuman primates . Memory T cells promote allograft rejection particularly in co-stimulation blockade-based immunosuppressive regimens . Here we show that the P06729 -specific fusion protein alefacept ( lymphocyte function-associated antigen-3-Ig ; LFA -3-Ig ) selectively eliminates memory T cells and , when combined with a co-stimulation blockade-based regimen using cytotoxic T lymphocyte antigen-4 ( P16410 ) -Ig , a P33681 - and P42081 -specific fusion protein , prevents renal allograft rejection and alloantibody formation in nonhuman primates . These results support the immediate translation of a regimen for the prevention of allograft rejection without the use of calcineurin inhibitors , steroids or pan-T cell depletion . Involvement of P27361 /2 , P47712 and NF-κB in microglia suppression by cannabinoid receptor agonists and antagonists . Cannabinoids have been consistently shown to suppress microglia activation and the release of cytotoxic factors including nitric oxide , superoxide and proinflammatory cytokines . However , the underlying molecular mechanisms and whether the action of cannabinoids is coupled to the activation of cannabinoid type 1 ( P21554 ) and type 2 ( CB2 ) receptors are still poorly defined . In this study we observed that the P21554 and CB2 receptor non-selective or selective agonists dramatically attenuate P35228 induction and ROS generation in LPS-activated microglia . These effects are due to their reduction of phosphorylation of extracellular signal regulated kinase 1/2 ( P27361 /2 ) , cytosolic phospholipase A ( cPLA ) and activation of NF-κB . Surprisingly , instead of reversing the effect of the respective P21554 and CB2 receptor agonists , the antagonists also suppress P35228 induction and ROS generation in activated microglia by similar mechanisms . Taken together , these results indicate that both cannabinoid receptor agonists and antagonists might suppress microglia activation by P21554 and CB2 receptor independent mechanisms , and provide a new insight into the mechanisms of microglia inhibition by cannabinoids . A pilot study evaluating genetic alterations that drive tobacco- and betel quid-associated oral cancer in Northeast India . The susceptibility of an individual to oral cancer is mediated by genetic factors and carcinogen-exposure behaviors such as betel quid chewing , tobacco use , and alcohol consumption . This pilot study was aimed to identify the genetic alteration in 100 bp upstream and downstream flanking regions in addition to the exonic regions of 169 cancer-associated genes by using Next Generation sequencing with aim to elucidate the molecular pathogenesis of tobacco- and betel quid-associated oral cancer of Northeast India . To understand the role of chemical compounds present in tobacco and betel quid associated with the progression of oral cancer , single nucleotide polymorphisms ( SNPs ) and insertion and deletion ( Indels ) found in this study were analyzed for their association with chemical compounds found in tobacco and betel quid using Comparative Toxogenomic Database . Genes ( AR , P38398 , P10145 , and P04637 ) with novel SNP were found to be associated with arecoline which is the major component of areca nut . Genes ( Q99728 , P51587 , P30279 , P08069 , P52701 , and Q9NS23 ) with novel deletion and genes ( P25054 , Q9HCU9 , O14519 , P42772 , P54826 , P08069 , and P06400 ) with novel insertion were found to be associated with aflatoxin B1 which is produced by fermented areca nut . Genes ( P28332 , P25054 , AR , Q99728 , Q9HCU9 , P38936 , Q01094 , P22455 , Q14315 , P01112 , P08069 , P29460 , P10145 , P41271 , P51692 , and P04637 ) with novel SNP were found to be associated with aflatoxin B1 . Genes ( Q13315 , P38398 , P38936 , P00533 , P10145 , and P04637 ) with novel SNP were found to be associated with tobacco specific nitrosamines . Recombinant P17936 inhibits allergic lung inflammation , P15692 production , and vascular leak in a mouse model of asthma . BACKGROUND : Vascular endothelial growth factor ( P15692 ) plays a pro-inflammatory mediator as well as a vascular permeability factor in bronchial asthma . P01308 -like growth factor ( IGF ) -I is also involved in the inflammatory process associated with bronchial asthma and stimulates P15692 expression . The IGF-binding proteins ( IGFBPs ) , especially P17936 , display distinctive properties and can interfere with various biological processes . METHODS : In this study , an ovalbumin ( OVA ) -induced murine model of allergic airway disease was used to investigate which mechanism is implicated in the preventive and therapeutic actions of P17936 administered exogenously on allergen-induced bronchial inflammation and airway hyper-responsiveness , in particular focusing on the regulation of P15692 expression . RESULTS : Administration of recombinant human P17936 to OVA-inhaled mice substantially attenuated the increases in hypoxia-inducible factor ( HIF ) -α activity , P05019 production , and P15692 protein levels in the lung . In addition , the blockade of P05019 action decreased the OVA-induced P15692 expression , airway inflammation , and bronchial hyper-responsiveness . The administration of recombinant human P17936 or CBO-P11 also reduced significantly increases in inflammatory cells , airway hyper-responsiveness , levels of P05112 , P05113 , P35225 , and vascular permeability in the lung of OVA-inhaled mice . Moreover , when recombinant human P17936 was administered after the completion of OVA inhalation , these therapeutic effects of P17936 were also observed . CONCLUSIONS : These results indicate that P17936 administered exogenously may attenuate antigen-induced airway inflammation and hyper-responsiveness through the modulation of vascular leakage and P15692 expression mediated by HIF-1α/HIF-2α signaling as well as P05019 action in allergic airway disease of mice . Essential role of p38 mitogen-activated protein kinase in contact hypersensitivity . The present study was designed to elucidate the role of p38 mitogen-activated protein kinase ( p38 ) in the pathogenesis of inflammation , using a mouse contact hypersensitivity ( Q99698 ) model induced by 2,4-dinitro-1-fluorobenzene ( DNFB ) . Ear swelling was induced by challenge with DNFB , accompanied by infiltration of mononuclear cells , neutrophils , and eosinophils and a marked increase in mRNA levels of cytokines such as interleukin ( IL ) -2 , interferon ( IFN ) -gamma , P05112 , P05113 , IL-1beta , Q14116 , and tumor necrosis factor-alpha in the challenged ear skin . Both ear swelling and the number of infiltrated cells in DNFB-challenged ear skin were significantly inhibited by treatment with SB202190 , a p38 inhibitor . Furthermore , the DNFB-induced expression of all cytokines except P05112 was significantly inhibited by treatment with SB202190 . Ribonuclease protection assay revealed that the mRNA levels of chemokines such as P02778 and P13500 in ear skin were markedly increased at 24 h after challenge with DNFB . The induction of these chemokines was significantly inhibited by treatment with SB202190 . In p38alpha +/- mice , both ear swelling and infiltration of cells induced by DNFB were reduced compared with those in wild-type mice . However , induction of cytokines by DNFB was also observed in p38alpha +/- mice , although the induction of P01579 , P05113 , and Q14116 was typically reduced compared with that in wild-type mice . Challenge with DNFB slightly induced P02778 and P13500 mRNA in p38alpha +/- mice , with weaker signals than those in SB202190-treated wild-type mice . These results suggest that p38 plays a key role in Q99698 and is an important target for the treatment of Q99698 . Topical glucocorticoids downregulate P23219 positive cells in nasal polyps . BACKGROUND : Influx of inflammatory cells is one of the hallmarks of nasal polyposis . As glucocorticoids ( GC ) are known to exhibit strong anti-inflammatory effects , these drugs are frequently used in the treatment of the disease . Part of the anti-inflammatory effects of GC is attributed to their interference with prostanoid synthesis . As cyclooxygenases ( P36551 ) are key enzymes in the synthesis of both pro- ( P23219 , P35354 ) and anti-inflammatory prostanoids ( P35354 ) , we investigated the role of topical GC on P23219 , P35354 and inflammatory markers in nasal polyps ( NP ) . METHODS : Immunohistochemical analysis of inflammatory markers ( P34810 , CD117 , MBP , elastase , IgE , BB-1 , P05112 , P05113 and P05231 ) , P23219 and P35354 was performed on normal nasal mucosa ( NM ) ( n = 18 ) , non-GC treated NP ( n = 27 ) and topical GC treated NP ( n = 12 ) . NP groups were matched for allergy , asthma and ASA intolerance . RESULTS : Increased numbers of eosinophils , P05113 + cells and IgE+ cells and decreased numbers of mastcells are striking features of NP inflammation ( P < 0.05 ) . In addition , increased numbers of P23219 + cells are observed in NP epithelium compared to NM ( P < 0.05 ) . CONCLUSION : Topical GC significantly reduce the number of P23219 + NP cells ( P < 0.05 ) , but have no significant effect on P35354 + NP cells . No significant reduction in the number of eosinophils is observed for GC treated NP . The number of P05113 + cells is however increased significantly upon GC treatment ( P < 0.05 ) . Effects of two Asian sand dusts transported from the dust source regions of Inner Mongolia and northeast China on murine lung eosinophilia . The quality and quantity of toxic materials adsorbed onto Asian sand dust ( P51689 ) are different based on dust source regions and passage routes . The aggravating effects of two ASDs ( ASD1 and ASD2 ) transported from the source regions of Inner Mongolia and northeast China on lung eosinophilia were compared to clarify the role of toxic materials in P51689 . The ASDs contained different amounts of lipopolysaccharides ( LPS ) and β-glucan ( ASD1 < ASD2 ) and SiO2 ( ASD1 > ASD2 ) . CD-1 mice were instilled intratracheally with ASD1 , ASD2 and/or ovalbumin ( OVA ) four times at 2-week intervals . ASD1 and ASD2 enhanced eosinophil recruitment induced by OVA in the submucosa of the airway , with goblet cell proliferation in the bronchial epithelium . ASD1 and ASD2 synergistically increased OVA-induced eosinophil-relevant cytokines interleukin-5 ( P05113 ) , P35225 ( ASD1 < ASD2 ) and chemokine eotaxin ( ASD1 > ASD2 ) in bronchoalveolar lavage fluid . ASD2 aggravating effects on lung eosinophilia were greater than ASD1 . The role of LPS and β-glucan in ASD2 on the production of pro-inflammatory mediators was assessed using in vitro bone marrow-derived macrophages ( BMDMs ) from wild type , O60603 -deficient ( O60603 -/- ) , O00206 -/- , and MyD88-/- mice ( on Balb/c background ) . ASD2-stimulated O60603 -/- BMDMs enhanced P05231 , IL-12 , P01375 -α , P13500 and MIP-1α secretion compared with ASD2-stimulated O00206 -/- BMDMs . Protein expression from ASD2-stimulated MyD88-/- BMDM were very low or undetectable . The in vitro results indicate that lung eosinophilia caused by P51689 is O00206 dependent . Therefore , the aggravation of OVA-related lung eosinophilia by P51689 may be dependent on toxic substances derived from microbes , such as LPS , rather than SiO2 . Analgesic effect of etoricoxib compared to ibuprofen on post endodontic pain . AIMS : DB01628 is a second-generation selective P35354 inhibitor . There are a few researches investigating analgesic effect of DB01628 in dentistry . METHODS : This randomized , double-blind , active-control study included sixty patients with clinical pulpal diagnosis of necrosis of the first mandibular molar and an associated periapical radiolucency who experienced severe pain ( more than 60 out of 100 in scale of Visual Analog Scale ( VAS ) . The patients were equally randomized into four groups , who received 60 mg etoricoxib ( group 1 ) , 90 mg etoricoxib ( group 2 ) , 120 mg etoricoxib ( group 3 ) , and 400 mg ibuprofen ( group 4 ) . All patients randomly received a single dose of the drug after the first session of the root canal therapy . Using VAS , the severity of pain was recorded 2 , 4 , 6 , 12 , 24 , 48 , and 72 hours after the drug was administered . RESULTS : Changing trends of pain over the time was significant for all groups ( P=0.003 ) . In addition , there was not a significant difference between various study arms ( P=0.146 ) . CONCLUSION : The results showed that ibuprofen had a comparable effect with various dosage of etoricoxib and may remain as the choice analgesic for dental pulpal pain . Chromosome 5q candidate genes in coeliac disease : genetic variation at P05112 , P05113 , P15248 , P35225 , Q9UHF5 and P04150 . Genetic predisposition to coeliac disease ( CD ) is determined primarily by alleles at the P01920 locus , and evidence exists implicating other major histocompatibility complex-linked genes ( 6p21 ) and the P16410 locus on chromosome 2q33 . In addition , extensive family studies have provided strong , reproducible evidence for a susceptibility locus on chromosome 5q ( CELIAC2 ) . However , the gene responsible has not been identified . We have assayed genetic variation at the P05112 , P05113 , P15248 , P35225 , Q9UHF5 and P04150 ( GR ) loci , all of which are present on chromosome 5q and have potential or demonstrated involvement in autoimmune and/or inflammatory disease , in a sample of 409 CD cases and 355 controls . Thirteen single nucleotide polymorphisms were chosen on the basis of functional relevance , prior disease association and , where possible , prior knowledge of the haplotype variation present in European populations . There were no statistically significant allele or haplotype frequency differences between cases and controls . Therefore , these results provide no evidence that these loci are associated with CD in this sample population . Genetically rescued tetrahydrobiopterin-depleted mice survive with hyperphenylalaninemia and region-specific monoaminergic abnormalities . One of the possibly mutated genes in DOPA-responsive dystonia ( DRD , Segawa 's disease ) is the gene encoding P30793 , which is the rate-limiting enzyme for tetrahydrobiopterin ( BH4 ) biosynthesis . Based on our findings on 6-pyruvoyltetrahydropterin synthase ( Q03393 ) gene-disrupted ( Pts(-/-) ) mice , we suggested that the amount of tyrosine hydroxylase ( TH ) protein in dopaminergic nerve terminals is regulated by the intracellular concentration of BH4 . In this present work , we rescued Pts(-/-) mice by transgenic introduction of human Q03393 cDNA under the control of the dopamine beta-hydroxylase promoter to examine regional differences in the sensitivity of dopaminergic neurons to BH4-insufficiency . The DPS-rescued ( Pts(-/-) , DPS ) mice showed severe hyperphenylalaninemia . Human Q03393 was efficiently expressed in noradrenergic regions but only in a small number of dopaminergic neurons . DB03886 and dopamine contents , and TH activity in the striatum were poorly restored compared with those in the midbrain . TH-immunoreactivity in the lateral region of the striatum was far weaker than that in the medial region or in the nucleus accumbens . We concluded that dopaminergic nerve terminals projecting to the lateral region of the striatum are the most sensitive to BH4-insufficiency . Biochemical and pathological changes in DPS-rescued mice were similar to those in human malignant hyperphenylalaninemia and DRD . P22303 antagonist potentiated insulin action in fed but not fasted state . The glucose disposal effect of insulin is doubled in response to a meal . This meal-induced insulin sensitization results from insulin acting on the liver , in the presence of a permissive hepatic parasympathetic feeding signal and elevated hepatic glutathione ( DB00143 ) , to release hepatic insulin-sensitizing substance ( HISS ) , a hormone that acts selectively on skeletal muscle to stimulate insulin-mediated glucose uptake . Blockade of the parasympathetic feeding signal to the liver , either through surgical denervation or atropine-mediated antagonism of hepatic muscarinic receptors , eliminates the HISS response , resulting in HISS-dependent insulin resistance ( HDIR ) and decreasing the response to insulin by approximately 55 % in the fed state . P01308 action in Sprague-Dawley rats , as determined with a rapidly sampled , transient euglycemic clamp in response to insulin ( 50 mU/kg ) , is decreased in a dose-dependent manner by atropine . In this study , we have used the ED75 atropine-induced model of HDIR . After a submaximal dose of atropine , potentiation of the remaining parasympathetic effect with the acetylcholinesterase antagonist neostigmine significantly restored postprandial insulin sensitization in a dose-dependent manner with peak effect at 0.1 microg/kg/min . DB01400 reversed the insulin resistance induced by partial fasting and partial muscarinic inhibition ( hepatic DB00143 levels are at fed levels ) , but not that induced by surgical hepatic denervation ( DB00143 normal , no nerve signal ) or 24-h fasting ( low DB00143 ) . No potentiation of the response to insulin by neostigmine occurred in normal , fed rats . The data suggest the use of either direct or indirectly acting cholinergic agonists for the treatment of impaired postprandial insulin sensitization . Development of drugs to target interactions between leukocytes and endothelial cells and treatment algorithms for inflammatory bowel diseases . Increased understanding of the pathogenesis of inflammatory bowel diseases ( IBDs ) has led to new therapeutic strategies . One of these is to target the molecules that regulate interactions between leukocytes and endothelial cells at sites of inflammation ( mainly leukocyte integrins and endothelial cell adhesion molecules of the immunoglobulin superfamily ) . These molecules have been validated as therapeutic targets for Q9UKU7 ; several have shown efficacy , and 2 have been approved by the Food and Drug Administration for treatment of Q9UKU7 . DB00108 , the first anti-integrin antibody tested for treatment of Q9UKU7 , blocks the α4 subunit . Although it is effective , its clinical use has been limited by its association with risk of progressive multifocal leukoencephalopathy . Other , allegedly more selective drugs that affect leukocyte recruitment in the gastrointestinal tract have been developed or are under investigation and could increase safety . These include vedolizumab and Q99217 181 ( antibodies against α4β7 ) , etrolizumab ( anti-β7 ) , and PF-00547659 ( anti-mucosal vascular addressin cell adhesion molecule 1 ) . Other agents have been developed to block α4 ( the small molecule AJM300 ) , P51686 ( the small molecule DB05005 -B ) , and P02778 ( the antibody eldelumab ) . We review the scientific rationale for inhibiting interactions between leukocytes and endothelial cells to reduce intestinal inflammation and analyze the clinical studies that have been performed to test these new molecules , with particular attention to safety . We propose an evidence-based clinical positioning of this class of drugs . Chronic DB00470 treatment increases DB02527 levels and DB02527 -dependent protein kinase activity in some rat brain regions . When Delta(9)-tetrahydrocannabinol ( Delta(9)-THC,15 mg/kg ) was injected intraperitoneally twice a day for 6 days , tolerance to its analgesic effect appeared to be complete . Chronic exposure to Delta(9)-THC caused a significant reduction in P21554 receptor binding in all brain areas that contain this receptor . Cannabinoid receptor density was markedly reduced in the cerebellum ( 52 % ) , hippocampus ( 40 % ) and globus pallidum ( 47 % ) compared to 30 % in the cortex and striatum . Chronic exposure enhanced the DB02527 pathway , as shown by the significant increase of DB02527 levels and PKA activity in the areas with receptor down-regulation ( cerebellum , striatum and cortex ) . We propose that the increase in DB02527 cascade is part of the biochemical basis of cannabinoid tolerance . Agonism at P41595 receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5-hydroxytryptamine2B ( P41595 ) receptors . To evaluate whether agonism at P41595 receptors is a phenomenon of the class of the ergolines , we studied P41595 receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC50 8.42 and 8.72 ) . DB01200 acted as a partial agonist ( pEC50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5-HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 receptors seems not to be a class effect of the ergolines . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane . Hsp27 regulates epithelial mesenchymal transition , metastasis , and circulating tumor cells in prostate cancer . Defining the mechanisms underlying metastatic progression of prostate cancer may lead to insights into how to decrease morbidity and mortality in this disease . An important determinant of metastasis is epithelial-to-mesenchymal transition ( EMT ) , and the mechanisms that control the process of EMT in cancer cells are still emerging . Here , we report that the molecular chaperone Hsp27 ( P04792 ) drives EMT in prostate cancer , whereas its attenuation reverses EMT and decreases cell migration , invasion , and matrix metalloproteinase activity . Mechanistically , silencing Hsp27 decreased P05231 -dependent P40763 phosphorylation , nuclear translocation , and P40763 binding to the Twist promoter , suggesting that Hsp27 is required for P05231 -mediated EMT via modulation of P40763 /Twist signaling . We observed a correlation between Hsp27 and Twist in patients with prostate cancer , with Hsp27 and Twist expression each elevated in high-grade prostate cancer tumors . Hsp27 inhibition by DB06094 , an antisense therapy currently in phase II trials , reduced tumor metastasis in a murine model of prostate cancer . More importantly , DB06094 treatment decreased the number of circulating tumor cells in patients with metastatic castration-resistant prostate cancer in a phase I clinical trial . Overall , this study defines Hsp27 as a critical regulator of P05231 -dependent and P05231 -independent EMT , validating this chaperone as a therapeutic target to treat metastatic prostate cancer . Single dose topical corticosteroid inhibits P05113 and P35225 in nasal lavage following grass pollen challenge . BACKGROUND : Nasal lavage is a noninvasive method of obtaining inflammatory exudates following nasal allergen challenge ( Q9C000 ) , and permits cells and released mediators to be evaluated . OBJECTIVE : To determine the effects of a single dose of topical steroid on eosinophils and levels of chemokines and cytokines in nasal lavage fluid following Q9C000 in patients with allergic rhinitis . METHODS : Patients with grass pollen seasonal allergic rhinitis ( n = 32 ) out of the allergy season received either nasal budesonide ( 100 microg per nostril ) or matched placebo before allergen challenge in a double blind two-way crossover design . A semi-automated mixed bead array system was employed to measure multiple chemokines and cytokines in small volumes ( 50 microl ) of nasal lavage supernatants . RESULTS : Following Q9C000 there was a rapid onset of nasal symptoms together with nasal eosinophilia , and the appearance of P05113 and P35225 in lavages between 4 and 8 h . Elevated levels of eotaxin , RANTES , P10145 and P13500 were also detected following allergen challenge . A single dose of nasal budesonide caused a decrease in symptoms ( P < 0.05 ) and nasal eosinophils ( P < 0.05 ) with selective abrogation of P05113 and P35225 responses ( P < 0.05 ) , but a lack of effect on levels of eotaxin , RANTES , P10145 and P13500 . CONCLUSION : This study suggests that a single dose of nasal steroid has the capacity to selectively abolish P05113 and P35225 responses following Q9C000 . This model should be convenient for testing novel anti-inflammatory and immunoregulatory agents intended for the treatment of allergic rhinitis . Inhibition of P51587 and DB01643 Synthase Creates Multidrug Sensitive Tumor Cells via the Induction of Combined " Complementary Lethality " . A high mutation rate leading to tumor cell heterogeneity is a driver of malignancy in human cancers . Paradoxically , however , genomic instability can also render tumors vulnerable to therapeutic attack . Thus , targeting DNA repair may induce an intolerable level of DNA damage in tumor cells . P51587 mediates homologous recombination repair , and P51587 polymorphisms increase cancer risk . However , tumors with P51587 mutations respond better to chemotherapy and are associated with improved patient prognosis . P04818 ( TS ) is also involved in DNA maintenance and generates cellular thymidylate . We determined that antisense downregulation of P51587 synergistically potentiated drugs with mechanisms of action related to P51587 function ( cisplatin , melphalan ) , a phenomenon we named " complementary lethality. " TS knockdown induced complementary lethality to TS-targeting drugs ( 5-FUdR and pemetrexed ) but not DNA cross-linking agents . Combined targeting of P51587 and TS induced complementary lethality to both DNA-damaging and TS-targeting agents , thus creating multidrug sensitive tumors . In addition , we demonstrated for the first time that simultaneous downregulation of both targets induced combined complementary lethality to multiple mechanistically different drugs in the same cell population . In this study , we propose and define the concept of " complementary lethality " and show that actively targeting P51587 and TS is of potential therapeutic benefit in multidrug treatment of human tumors . This work has contributed to the development of a P51587 -targeting antisense oligdeoxynucleotide ( ASO ) " BR-1 " which we will test in vivo in combination with our TS-targeting ASO " Q8N1L9 83 " and attempt early clinical trials in the future.Molecular Therapy - Nucleic Acids ( 2013 ) 2 , e78 ; doi:10.1038/mtna.2013.7 published online 12 March 2013 .

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Structure-function studies of linear and cyclized peptide antagonists of the P30968 . Structurally new analogs of the peptidic P30968 antagonist DB00050 as well as conformationally constrained cyclized deca- or pentapeptides were synthesized and selected peptides evaluated comprehensively . To understand how structural variations of the antagonistic peptide effect pharmacodynamic properties , binding affinities and antagonistic potencies toward the human and rat P30968 were determined . Whereas large substituents in position 6 of linear peptides are compatible with high binding affinity ( K(D) < 0.5 nM ) , all cyclized peptides except the cyclo[3-10] analog D-52391 depicted low binding affinity ( K(D) > 10 nM ) . Binding affinity and antagonistic potency in vitro correlated for all peptides and surprisingly no discrimination between human and rat receptor proteins was observed . Since receptor residues W(101) and N(102) are involved in agonist and antagonist binding , equally potent but structurally different antagonists were tested for binding to the respective W(101)A and N(102)A mutants . In contrast to linear decapeptides , residues N(102) and W(101) are not involved in binding of D-23938 and W(101) is the critical residue for D-52391 binding . We conclude that although equally potent , peptidic P30968 antagonists do have distinct interactions within the ligand binding pocket . Finally , selected antagonists were tested for testosterone suppression in male rats . The duration of testosterone suppression below castration levels differed largely from 1 day for DB06785 to 27 days for D-23487 . Systemic availability became evident as the most important parameter for in vivo efficacy . A field synopsis and meta-analysis of genetic association studies in peripheral arterial disease : The CUMAGAS-PAD database . In an electronic search of the literature , the authors systematically retrieved all published studies that investigated genetic susceptibility to peripheral arterial disease ( PAD ) . They created a comprehensive database of all eligible studies , collecting detailed genetic and bioinformatics data on each polymorphism . Data from eligible studies were synthesized using meta-analysis techniques . Gene variants were classified into distinct pathophysiologic pathways , and their potential involvement in PAD pathogenesis was determined . Forty-one publications that examined 44 gene polymorphisms were included . For 37 polymorphisms , the variant form had a functional effect . Twenty-three polymorphisms in 22 potential PAD candidate genes ( F2 , P02675 , P42898 , P05106 , P12821 , AGT , P05231 , P13500 , P05362 , P16581 , P14780 , P37231 , P03956 , P35611 , Q9H244 , P11150 , Q13093 , Q8WTV0 , P08254 , P55157 , P08519 , P32297 ) showed a significant association in individual studies . Eighty-eight percent of the studies had statistical power of less than 50 % , and in 15 studies the genotype distribution in the control group did not conform to Hardy-Weinberg equilibrium . Data on 12 polymorphisms ( P12259 1691 G/A , P42898 677C/T , F2 20210 G/A , P05106 1565 T/C , P12821 I/D , AGT 704C/T , AGT -6G/A , AGT 733C/T , P05231 -174 G/C , P14780 -1562C/T , P05362 1462A/G , P32297 831C/T ) were synthesized , and a positive association was found for 3 ( P05231 -174 G/C , P05362 1462A/G , P32297 831C/T ) . DB00114 values in cerebrospinal fluid : reference values and diagnosis of Q9NVS9 deficiency in paediatric patients . Our aim was to establish reference values for cerebrospinal fluid ( P04141 ) pyridoxal 5'-phosphate ( PLP ) in a paediatric population for the diagnosis of pyridox(am)ine 5'-phosphate oxidase ( Q9NVS9 ) deficiency . For reference values , P04141 samples from 113 paediatric controls ( age range : 1 day-18 years ) from Barcelona and London were analysed . Cerebrospinal fluid PLP and biogenic amine concentrations were analysed by HPLC with fluorescence and electrochemical detection . DB00114 concentrations in 4 patients with Q9NVS9 deficiency were determined . A negative correlation between P04141 PLP values and age of controls was observed in both populations ( r=-0.503 ; p < 0.0001 and r=-0.542 ; p=0.002 ) . Reference values were stratified into 4 ( Barcelona ) and 3 age groups ( London ) . For the newborn period , P04141 PLP reference intervals were 32-78 and 44-89 nmol/L for the Barcelona and London centers , respectively ) . No correlation was observed in the different age groups between PLP values and biogenic amines metabolites . PLP values in neonates with Q9NVS9 deficiency were clearly decreased ( PLP=3.6 , 12.0 , 14.0 and 18.0 nmol/L ) compared with our reference ranges . In conclusion , reference values for P04141 PLP should be stratified according to age . No association was observed between PLP values and biogenic amines metabolites . In our 4 cases with Q9NVS9 deficiency , P04141 PLP values were clearly below the reference values . Anti-interleukin 6 receptor antibody treatment in rheumatic disease . Interleukin 6 ( P05231 ) is a pleiotropic cytokine with a wide range of biological activities . P05231 transgene into mice gives rise to the abnormalities such as hypergammaglobulinaemia , thrombocytosis , infiltration of inflammatory cells into the tissues , mesangial cell proliferation of the kidney as well as splenomegaly and lymphadenopathy , which are predictable by the biological functions of P05231 shown in vitro . Continuous overproduction of P05231 is observed in patients with some immune-inflammatory diseases such as Castleman 's disease and rheumatoid arthritis that are frequently associated with similar abnormalities to those of P05231 transgenic mice , strongly suggesting the involvement of P05231 in the human diseases . Successful treatment of the model animals for immune-inflammatory diseases with anti- P05231 receptor ( P08887 ) antibody thus indicates the possible application of P05231 blocking agents to treat the P05231 related immune-inflammatory diseases of humans . In this review , the new therapeutic strategy for Castleman 's disease and RA using humanized antibody to human P05231 receptor , DB06273 , is discussed . Maturation of dendritic cells by recombinant human P29965 -trimer leads to a homogeneous cell population with enhanced surface marker expression and increased cytokine production . Dendritic cells ( DC ) have been shown to be potent inducers of specific cytotoxic T-cell responses both in vivo and in vitro . Furthermore , exposure to cytokines such as tumour necrosis factor ( P01375 ) -alpha or P25942 triggering changes DC phenotype and cytokine production and may enhance the T-cell activating capacity of the DC . We studied DC phenotype and cytokine production as well as the T-cell proliferation and cytotoxic T lympocyte ( CTL ) activation induced by DC generated in vitro . In addition , the effect of exposure to recombinant human P29965 -trimer ( huCD40LT ) on these parameters was investigated . Effective differentiation of monocytes derived from freshly isolated peripheral blood mononuclear cells ( PBMC ) was obtained with granulocyte macrophage-colony stimulating factor ( GM- P04141 ) and interleukin ( IL ) -4 . The DC expression of human leucocyte antigen ( HLA ) molecules , P33681 , Q01151 , and P42081 was markedly enhanced by exposure to huCD40LT even compared to P01375 exposure . Only a moderate cytokine production was observed initially , while P01375 addition or P25942 triggering , especially , induced enhanced production of P05231 and IL-12 p40 . Surprisingly , comparable induction of T-cell proliferation by a DC allostimulus or through the presentation of PPD , and influenza M1-peptide specific CTL activity was obtained with nonmaturated ( Q01151 - ) and maturated ( Q01151 + ) DC . In conclusion , a final maturation of monocyte-derived DC through huCD40LT resulted in a highly homogeneous cell population with enhanced surface marker expression and high production of pro-inflammatory cytokines . In addition , the induction of responses to allo or recall antigens presented by huCD40LT maturated DC was comparable to the responses obtained with the DC maturated through P01375 exposure . Characterization of the effect of chronic administration of a calcium-sensing receptor antagonist , DB05255 , on renal calcium excretion and serum calcium in postmenopausal women . Ronacaleret is an orally-active calcium-sensing receptor ( P41180 ) antagonist that has the potential for therapeutic utility in the stimulation of PTH release , notably as a bone anabolic agent comparable to recombinant human PTH(1-34) ( DB05829 (1-34) ) . A recent study has shown that , despite the ability to increase circulating PTH levels in postmenopausal women in a dose-dependent manner , minimal effects of DB05255 on bone mineral density have been observed . Therefore , the purpose of this study was to characterize the PTH profile as well as calcium metabolism parameters as a marker of PTH biological activity following the administration of DB05255 or DB05829 (1-34) . Administration of DB05255 led to lower peak levels of PTH than were observed with DB05829 (1-34) , however , greater total PTH exposure was observed . Further , chronic administration of either agent was associated with increases in urinary calcium excretion and serum calcium levels , with the magnitude of the changes following DB05255 significantly greater than that for DB05829 (1-34) . The greater magnitude of effects observed with DB05255 is likely due to the greater total PTH exposure , and is potentially reflective of a state comparable to mild hyperparathyroidism . It is not clear whether the administration of all calcilytics would lead to a similar result , or is due to characteristics specific to DB05255 . Human plasmacytoid dendritic cells express an atrial natriuretic peptide receptor , guanylyl cyclase-A . Atrial natriuretic peptide is a cardiovascular hormone secreted mainly by the cardiac atria and regulates the volume-pressure homeostasis . The action of P01160 is mediated by P16066 . We previously reported that human monocyte-derived dendritic cells express P16066 and respond to P01160 with polarization toward a Th2-inducing phenotype . In the present study , we explored the possibility that pDC are subjected to immunoregulation via the P01160 / P16066 system . We examined P16066 expression on blood pDC and found that P16066 was not expressed on fresh pDC but was induced after stimulation with CpG-oligodeoxynucleotide AAC-30 , P08700 , or interleukin-3 plus P29965 . Activated pDC responded to P01160 with an increase in cGMP production , indicating that P16066 expressed on pDC was functional . We investigated whether tonsillar pDC express P16066 by immunohistochemistry and immunofluorescence staining . We found that P16066 (+) HLA-DR(+) cells were present in the T-cell areas and the perivascular areas . Flow cytometric analysis with tonsillar cells confirmed that lineage(-) CD123(high) pDC express P16066 . These results indicate that the P01160 / P16066 system is involved in immune regulation through pDC in secondary lymphoid organs . Solution structure and backbone dynamics of the catalytic domain of matrix metalloproteinase-2 complexed with a hydroxamic acid inhibitor . P08253 is a member of the matrix metalloproteinase family that has been implicated in tumor cell metastasis and angiogenesis . Here , we describe the solution structure of a catalytic domain of P08253 complexed with a hydroxamic acid inhibitor ( DB01630 ) , determined by three-dimensional heteronuclear NMR spectroscopy . The catalytic domain , designated MMP-2C , has a short peptide linker replacing the internal fibronectin-domain insertion and is enzymatically active . Distance geometry-simulated annealing calculations yielded 14 converged structures with atomic root-mean-square deviations ( r.m.s.d. ) of 1.02 and 1.62 A from the mean coordinate positions for the backbone and for all heavy atoms , respectively , when 11 residues at the N-terminus are excluded . The structure has the same global fold as observed for other MMP catalytic domains and is similar to previously solved crystal structures of P08253 . Differences observed between the solution and the crystal structures , near the bottom of the S1 ' specificity loop , appear to be induced by the large inhibitor present in the solution structure . The MMP-2C solution structure is compared with P22894 crystal structure bound to the same inhibitor to highlight the differences especially in the S1 ' specificity loop . The finding provides a structural explanation for the selectivity between P08253 and P22894 that is achieved by large inhibitors . Intraplatelet signaling mechanisms of the priming effect of matrix metalloproteinase-2 on platelet aggregation . OBJECTIVE : Platelets contain and release some matrix metalloproteinases ( MMPs ) , enzymes involved in the degradation of extracellular matrix , and one of these ( P08253 ) exerts a proaggregatory effect . We explored the signal transduction mechanisms activated by P08253 in human blood platelets . METHODS AND RESULTS : Recombinant , human P08253 , added before stimulation with subthreshold doses of different agonists , potentiated platelet activation , calcium influx , IP3 formation , and pleckstrin phosphorylation . Wortmannin and LY29400 , two P19957 -K inhibitors , suppressed the potentiating effects of P08253 and preincubation with P08253 enhanced the thrombin-induced association of the p85alpha P19957 -K subunit with the cytoskeleton and increased the phosphorylation of P31749 . Protein tyrosine kinase inhibitors , Q96HU1 kinase inhibitors , P04054 inhibitors , cyclooxygenase inhibitors and antagonists of the P47900 and Q9H244 receptors did not affect the potentiating activity of P08253 on platelets . CONCLUSION : Our data show that P08253 , at a concentration released by activated platelets , facilitates platelet activation acting at the level of a second messenger system common to different agonists and related to the activation of P19957 -K . Platelet-released P08253 may contribute to platelet activation in vivo . Aurora kinase inhibitors reveal mechanisms of Q15398 in nucleation of centrosomal and kinetochore microtubules . The overexpression of Aurora kinases in multiple tumors makes these kinases appealing targets for the development of anticancer therapies . This study identified two small molecules with a furanopyrimidine core , IBPR001 and IBPR002 , that target Aurora kinases and induce a DFG conformation change at the DB00171 site of Aurora A . Our results demonstrate the high potency of the IBPR compounds in reducing tumorigenesis in a colorectal cancer xenograft model in athymic nude mice . Human hepatoma up-regulated protein ( Q15398 ) is a substrate of O14965 , which plays a crucial role in the stabilization of kinetochore fibers . This study used the IBPR compounds as well as DB05220 , a proven Aurora A inhibitor , as chemical probes to investigate the molecular role of Q15398 in mitotic spindle formation . These compounds effectively eliminated Q15398 phosphorylation , thereby revealing the coexistence and continuous cycling of Q15398 between unphosphorylated and phosphorylated forms that are associated , respectively , with microtubules emanating from centrosomes and kinetochores . Furthermore , these compounds demonstrate a spatial hierarchical preference for Q15398 in the attachment of microtubules extending from the mother to the daughter centrosome . The finding of inequality in the centrosomal microtubules revealed by these small molecules provides a versatile tool for the discovery of new cell-division molecules for the development of antitumor drugs . P30968 and peritoneal plasmin activity . Most surgical procedures performed by obstetrician-gynecologists are associated with pelvic adhesions that cause subsequent serious sequelae , including small bowel obstruction , infertility , chronic pelvic pain , and difficulty in postoperative treatment , including complexity during subsequent surgical procedures . This study was conducted to determine if gonadotropin-releasing hormone analogues ( GnRHa ) affect the expressing tissue-type plasminogen activator ( t-PA ) and its inhibitor-1 ( P05121 ) in peritoneal cells in culture . Human peritoneal Met5A cells were used to examine the effects of GnRHa leuprolide , buserelin and goserelin on the levels of t-PA and PA-1 . Antigen concentrations were measured in conditioned media and cell lysates by real-time PCR and ELISA . P30968 ( GnRHR ) mRNA was determined by RT-PCR . GnRHR mRNA was detected in Met5A cells . Exposure of Met5A cells to GnRHa induced a rapid decrease of P05121 level in cultured medium but not in cell lysate ( protein and mRNA ) . These effects of GnRHa on P05121 were not associated with any changes in t-PA level . These results suggest that GnRHa may be an effective stimulator of local peritoneal fibrinolytic activity , as it decreases P05121 secretion in peritoneal Met5A cells by a mechanism linked to GnRHR . Lack of direct action of atriopeptidase inhibitor on cellular pH regulation in rabbit S2 proximal straight tubules . While the in vivo diuretic and antihypertensive effects of the newly-developed drug DB00616 ( UK-79300 ) , a prodrug of a selective atriopeptidase inhibitor UK-73967 ( DB00058 ) are well established , the mechanism of its diuretic action is not yet fully understood due to the lack of information about its direct effects on proximal tubules . To elucidate whether this atriopeptidase inhibitor has any direct effects on proximal tubules , we examined the effect of DB00058 on intracellular pH ( pHi ) of the in vitro microperfused proximal S2 segment of rabbit kidney , using the pH-sensitive fluorescent dye , (2',7')-bis(carboxyethyl)-(5,6)-carboxyfluorescein ( BCECF ) . In the steady-state condition , pHi was 7.13 +/- 0.02 ( n = 19 ) . P08473 inhibitor ( DB00058 ) at 100 microM added to the bath changed pHi by only 0.02 +/- 0.07 unit ( n = 7 , p > 0.05 ) in 10 min . The same concentration of DB00058 in the lumen changed pHi by 0.03 +/- 0.02 unit ( n = 6 , p > 0.05 ) . To test whether the synergistic effects of DB00058 on the luminal and basolateral transport systems prevented the apparent change in pHi , we also examined the effect of DB00058 in the presence of amiloride in the lumen . The inhibition of Na+/H+ antiporter by the addition of 1 mM of amiloride to the lumen caused no change in pHi response to basolateral DB00058 application . DB00058 was without effect on pHi also in the presence of atrial natriuretic polypeptide ( P01160 ) . These results suggest that DB00058 per se has no significant effect on the process of proximal acidification over a short time period . DB08816 reduces neutrophil recruitment and lung damage in abdominal sepsis . Abstract Platelets play an important role in abdominal sepsis and Q9H244 receptor antagonists have been reported to exert anti-inflammatory effects . Herein , we assessed the impact of platelet inhibition with the Q9H244 receptor antagonist ticagrelor on pulmonary neutrophil recruitment and tissue damage in a model of abdominal sepsis . Wild-type C57BL/6 mice were subjected to cecal ligation and puncture ( CLP ) . Animals were treated with ticagrelor ( 100 mg/kg ) or vehicle prior to CLP induction . Edema formation and bronchoalveolar neutrophils as well as lung damage were quantified . Flow cytometry was used to determine expression of platelet-neutrophil aggregates , neutrophil activation and P29965 expression on platelets . CLP-induced pulmonary infiltration of neutrophils at 24 hours was reduced by 50 % in ticagrelor-treated animals . Moreover , ticagrelor abolished CLP-provoked lung edema and decreased lung damage score by 41 % . Notably , ticagrelor completely inhibited formation of platelet-neutrophil aggregates and markedly reduced thrombocytopenia in CLP animals . In addition , ticagrelor reduced platelet shedding of P29965 in septic mice . Our data indicate that ticagrelor can reduce CLP-induced pulmonary neutrophil recruitment and lung damage suggesting a potential role for platelet antagonists , such as ticagrelor , in the management of patients with abdominal sepsis . The death domain kinase Q13546 links the immunoregulatory P25942 receptor to apoptotic signaling in carcinomas . P25942 , a tumor necrosis factor ( P01375 ) receptor family member , is widely recognized for its prominent role in the antitumor immune response . The immunostimulatory effects of P25942 ligation on malignant cells can be switched to apoptosis upon disruption of survival signals transduced by the binding of the adaptor protein Q9Y4K3 to P25942 . Apoptosis induction requires a TRAF2-interacting P25942 motif but is initiated within a cytosolic death-inducing signaling complex after mobilization of receptor-bound TRAF2 to the cytoplasm . We demonstrate that receptor-interacting protein 1 ( Q13546 ) is an integral component of this complex and is required for P29965 -induced caspase-8 activation and tumor cell killing . Degradation of the Q13546 K63 ubiquitin ligases cIAP1/2 amplifies the P25942 -mediated cytotoxic effect , whereas inhibition of Q9NQC7 , a Q13546 K63 deubiquitinating enzyme , reduces it . This two-step mechanism of apoptosis induction expands our appreciation of commonalities in apoptosis regulatory pathways across the P01375 receptor superfamily and provides a telling example of how P01375 family receptors usurp alternative programs to fulfill distinct cellular functions . [ Prominent features of management strategies in acute coronary syndromes with the new oral antiplatelet agents ] . The novel oral Q9H244 inhibitors ( prasugrel and ticagrelor ) have been incorporated into the recently updated acute coronary syndrome ( ACS ) guidelines , as an adjunct antiplatelet treatment to aspirin . The studies involving the use of new oral antiplatelet agents that are more potent , predictable and faster platelet inhibitors than clopidogrel have demonstrated superiority with respect to the primary composite endpoint ( cardiovascular death , non-lethal myocardial infarction , stroke ) for both prasugrel and ticagrelor compared to clopidogrel . The subgroup analysis of the relevant studies showed that these new agents differ in their level of efficacy in different ACS patient subgroups : ( 1 ) Mortality was reduced with ticagrelor ; ( 2 ) DB08816 is especially more effective in intermediate-and high-risk non-ST elevation ACS patients in whom early invasive strategy is selected ; ( 3 ) Prasugrel should be especially preferred in patients with acute ST elevation myocardial infarction undergoing percutaneous coronary intervention ( P05154 ) after diagnostic angiography ; and ( 4 ) Prasugrel is more effective in diabetic patients . While clopidogrel is recommended for ACS patients who are followed with a non-invasive strategy or who have not undergone percutaneous revascularization , it is the last line choice or an alternative to the Q9H244 inhibitor therapy for patients undergoing invasive strategy . G protein-coupled receptors disrupted in human genetic disease . Genetic variation in G protein-coupled receptors ( GPCRs ) results in the disruption of GPCR function in a wide variety of human genetic diseases . In vitro strategies have been used to elucidate the molecular pathologies that underlie naturally occurring GPCR mutations . Various degrees of inactive , overactive , or constitutively active receptors have been identified . These mutations often alter ligand binding , G protein coupling , receptor desensitization , and receptor recycling . The role of inactivating and activating calcium-sensing receptor ( P41180 ) mutations is discussed with respect to familial hypocalciuric hypercalemia ( FHH ) and autosomal dominant hypocalemia ( DB00067 ) . Among DB00067 mutations , those associated with tonic-clonic seizures are discussed . Other receptors discussed include rhodopsin , thyrotropin , parathyroid hormone , melanocortin , follicle-stimulating hormone , luteinizing hormone , gonadotropin-releasing hormone ( GnRHR ) , adrenocorticotropic hormone , vasopressin , endothelin-beta , purinergic , and the G protein associated with asthma ( Q6W5P4 ) . Diseases caused by mutations that disrupt GPCR function are significant because they might be selectively targeted by drugs that rescue altered receptors . Examples of drug development based on targeting GPCRs mutated in disease include the calcimimetics used to compensate for some P41180 mutations , obesity therapeutics targeting melanocortin receptors , interventions that alter GnRHR loss from the cell surface in idiopathic hypogonadotropic hypogonadism and novel drugs that might rescue the Q9H244 receptor in a rare bleeding disorder . The discovery of Q6W5P4 suggests that drug screens against variant GPCRs may identify novel drugs . This review of the variety of GPCRs that are disrupted in monogenic disease provides the basis for examining the significance of common pharmacogenetic variants . Q9H244 receptor signalling towards P31749 proceeds through P08069 cross-talk and requires activation of Src , Pyk2 and Rap1 . Previously it was shown that stimulation of the Q9H244 receptor activates P31749 signalling in P13671 glioma cells [ K. Van Kolen and H. Slegers , J. Neurochem. 89 , 442. ] . In the present study , the mechanisms involved in this response were further elucidated . In cells transfected with the Gbetagamma-scavenger beta- O14965 / P25098 or Rap1GAPII , stimulation with 2MeSADP failed to enhance P31749 phosphorylation demonstrating that the signalling proceeds through Gbetagamma-subunits and Rap1 . Moreover , Rap1-GTP pull-down assays revealed that Q9H244 receptor stimulation induced a rapid activation of Rap1 . Treatment of cells with the Ca2+ chelator BAPTA-AM and inhibition of Src and O14939 with Q99463 or 1-butanol , respectively , abrogated Q9H244 receptor-mediated activation of Rap1 and P31749 . In addition inhibition of PKCzeta decreased basal and 2MeSADP-stimulated phosphorylation of P31749 indicating a role for this PKC isoform in P31749 signalling . Although the increased P31749 phosphorylation was abolished in the presence of the P08069 tyrosine kinase inhibitor AG 1024 , 2MeSADP did not significantly increase receptor phosphorylation . Nevertheless , phosphorylation of a 120 kDa P08069 -associated protein was observed . The latter protein was identified by MALDI-TOF/TOF-MS as the proline-rich tyrosine kinase 2 ( Pyk2 ) that co-operates with Src in a O14939 -dependent manner . Consistent with the signalling towards Rap1 and P31749 , activation of Pyk2 was abrogated by Ca2+ chelation , inhibition of O14939 and P08069 tyrosine kinase activity . In conclusion , the data reveal a novel type of cross-talk between Q9H244 and P05019 receptors that proceeds through Gbetagamma- , Ca2+-and O14939 -dependent activation of the Pyk2/Src pathway resulting in GTP-loading of Rap1 required for an increased P31749 phosphorylation . Glycoprotein IIb/IIIa and Q9H244 receptor antagonists yield additive inhibition of platelet aggregation , granule secretion , soluble P29965 release and procoagulant responses . Glycoprotein IIb/IIIa ( P08514 /IIIa ) antagonists , including abciximab and tirofiban , are administered concurrently with clopidogrel , a Q9H244 antagonist , and aspirin in some patients undergoing percutaneous coronary intervention . We studied the effects of , and interactions between , abciximab , tirofiban , aspirin and the Q9H244 antagonist cangrelor on platelet aggregation , alpha and dense granule secretion and procoagulant responses in vitro . Blood was obtained from healthy volunteers . Platelet aggregation , dense granule secretion , alpha granule secretion ( P05121 and soluble P29965 levels ) and procoagulant responses ( annexin-V and microparticle formation ) were assessed using collagen and thrombin receptor activating peptide ( TRAP ) as agonists . All the antagonists used singularly inhibited collagen-induced responses . Combinations of abciximab or tirofiban with aspirin and/or cangrelor gave additive inhibition with the greatest effect seen when abciximab or tirofiban was combined with both aspirin and cangrelor . DB06441 inhibited TRAP-induced responses and , again , there was additive inhibition of these parameters when abciximab or tirofiban were combined with cangrelor . The P08514 /IIIa receptor plays an important role in amplification of platelet activation such that there are important interactions between P08514 /IIIa antagonists and inhibitors of both Q9H244 receptor activation and , to a lesser extent , thromboxane A2 generation . These interactions are likely to have important influences on the safety and efficacy of combination anti-platelet therapies . Mutational analysis of the mitochondrial P47985 of Saccharomyces cerevisiae . II . Biochemical characterization of temperature-sensitive Q13546 - mutations . Although the function of the P47985 is generally understood , little is known of how the structure of this protein supports its mechanistic role in electron transfer in the cytochrome bc1 complex . To better understand the structural basis of iron-sulfur protein function , we have undertaken a mutational analysis of the gene encoding this protein and initially isolated five temperature-sensitive iron-sulfur protein mutants ( Beckmann , J. D. , Ljungdahl , P. O. , and Trumpower , B. L. ( 1989 ) J. Biol. Chem. 264 , 3713-3722 ) . Each of the five ts-rip1- mutants exhibited pleiotropic effects . Although the mutant iron-sulfur proteins manifest several in vitro phenotypes in common , each exhibited unique characteristics . All of the ts-rip1- mutations resulted in membranes with decreased ubiquinol-cytochrome c oxidoreductase activities and decreased thermostability compared to membranes containing wild type iron-sulfur protein . All of the mutations conferred slight but significant resistance to the respiratory inhibitor myxothiazol , and one mutant was hypersensitive to inhibition by DB04799 , a structural analog of ubiquinone . In addition , one of the mutations completely blocks post-translational processing of the iron-sulfur protein , leading to accumulation of pre-iron-sulfur protein in mitochondrial membranes at nonpermissive temperatures . Finally , a mutation 12-amino acid residues away from the carboxyl terminus ( 203S ) results in an extremely unstable protein . This region of the protein may be essential in blocking degradation of pre-iron-sulfur protein by cytoplasmic proteases as the protein is imported into the mitochondria , or may be a " degradation signal , " which tags the iron-sulfur protein for turnover . P04271 modulates growth factors and costimulatory molecules expression in cultured human astrocytes . P04271 is a Ca(2+)-binding protein expressed in the nervous system . Increased levels of P04271 in the extracellular space have been detected in several neurological disorders . We investigated the response of human astrocytes to micromolar chronic concentrations of this protein measuring the expression of some costimulatory molecules , such as Q07011 , Q07011 -L , P25942 , P29965 , the chemokine RANTES and two growth factors P09038 and TGF-β2 . Our findings suggest that high levels of P04271 in the brain parenchyma may modulate the activation status of astrocytes decreasing their neuroprotective role and modifying their interaction with microglia and other inflammatory cells . This effect may contribute to evolution of some neurological disorders . DB04864 , but not tacrine , stimulates P04271 secretion in astrocyte cultures . AIMS : The loss of cholinergic function in the central nervous system contributes significantly to the cognitive decline associated with advanced age and dementias . DB04864 ( HupA ) is a selective inhibitor of acetylcholinesterase ( P22303 ) and has been shown to significantly reduce cognitive impairment in animal models of dementia . Based on the importance of astrocytes in physiological and pathological brain activities , we investigated the effect of HupA and tacrine on P04271 secretion in primary astrocyte cultures . P04271 is an astrocyte-derived protein that has been proposed to be a marker of brain injury . MAIN METHODS : Primary astrocyte cultures were exposed to HupA , tacrine , cholinergic agonists , and P04271 secretion was measured by enzyme-linked immunosorbent assay ( ELISA ) at 1 and 24h . KEY FINDINGS : HupA , but not tacrine , at 100μM significantly increased P04271 secretion in astrocyte cultures . DB00184 ( at 100 and 1000μM ) was able to stimulate P04271 secretion in astrocyte cultures . SIGNIFICANCE : Our data reinforce the idea that P22303 inhibitors , particularly HupA , do not act exclusively on the acetylcholine balance . This effect of HupA could contribute to improve the cognitive deficit observed in patients , which are attributed to cholinergic dysfunction . In addition , for the first time , to our knowledge , these data indicate that P04271 secretion can be modulated by nicotinic receptors , in addition to glutamate , dopamine and serotonin receptors .

MH_train_1325

MH_train_1325

MH_train_1325

interacts_with DB00977?

multiple_choice

Inhibitory effect of natriuretic peptides on aldosterone synthase gene expression in cultured neonatal rat cardiocytes . BACKGROUND : Although previously thought to be synthesized solely in adrenal cortex , we have recently showed that aldosterone is also produced in and the expression of P19099 mRNA was induced in the failing or hypertensive human ventricle . Atrial natriuretic peptide ( P01160 ) and brain natriuretic peptide ( DB04899 ) are cardiac hormones with wide biological effects , including inhibition of renin and aldosterone production . We hypothesized that natriuretic peptides reduce the expression of P19099 mRNA in the heart . METHODS AND RESULTS : To test this hypothesis , we examined whether endogenous or exogenous natriuretic peptides reduce the expression of P19099 mRNA using real-time reverse transcription-polymerase chain reaction . By using HS 142-1 , a functional guanylyl cyclase-A type receptor antagonist , we showed that angiotensin II ( AngII ) pretreated with HS 142-1 increased P19099 mRNA expression ( 1.62+/-0.12-fold , HS 142-1+AngII 10(-7) mol/L versus AngII 10(-7) mol/L alone , P < 0.0001 ) . The treatment with exogenous ( 10(-6) mol/L ) P01160 and DB04899 reduced P19099 mRNA expression ( P01160 , P=0.0042 ; DB04899 , P=0.0012 ) . CONCLUSIONS : We showed that endogenous and exogenous natriuretic peptides reduced P19099 mRNA expression in cultured neonatal rat cardiocytes . This may inhibit the cardiac renin-angiotensin-aldosterone system by suppressing the gene expression of P19099 and restraining cardiac hypertrophy and fibrosis . Increased blood pressure and loss of anp-induced natriuresis in mice lacking Q9UD71 gene . Atrial natriuretic peptide ( P01160 ) is an important regulator of sodium metabolism and indirectly of blood pressure . Evidence has accumulated that P01160 regulates sodium metabolism through a cascade of steps involving an increase in the level of cGMP , activation of cGMP-dependent protein kinase ( PKG ) , and inhibition of renal tubular Na+ , K+-ATPase activity . One of the major substrates for PKG is Q9UD71 . In the present study we observed that P01160 does not induce natriuresis in mice that lack Q9UD71 . In contrast , there was a 4-fold increase in urinary sodium excretion following P01160 administration to wild type mice . P01160 as well as Zaprinast , a selective inhibitor of cGMP phosophodiesterase , inhibited renal Na+ , K+-ATPase activity in wild type mice but had no such effect in mice lacking Q9UD71 . Mean arterial blood pressure , measured in conscious animals , was significantly increased in Q9UD71 deficient mice as compared to wild type mice . The results confirm that Q9UD71 acts as a third messenger in the P01160 signaling pathway in renal tissue and suggest an important role of Q9UD71 in the maintenance of normal blood pressure . DB08875 as a novel therapy for renal cell carcinoma . DB08875 / DB05153 ( Exelexis , Inc. ) has demonstrated remarkable responses in kidney cancer . Preclinical results revealed P15692 , P10721 and MET inhibition in a variety of solid tumors such as thyroid , ovarian , renal , lung , liver and prostate cancers . A phase II trial demonstrated efficacy in renal cancer with a 28 % objective response rate , stable disease rate of 62 % and median progression free survival of 14.7 months . Predominant toxicities of fatigue and diarrhea were noted . Dramatic responses in bone metastases ( three of four patients ) make the agent especially valuable for palliation in a disease , where presence of bone metastases is a predictor of worse survival . DB08875 is an emerging novel agent with promising activity in advanced kidney cancer . Randomized trials are planned in comparison with standard P15692 inhibitor therapy . Defining the role of MET overexpression would help patient selection and enrich and enhance the future evaluation of this targeted novel agent . DB00741 response to stress is associated with myocardial remodeling in salmonid fishes . Cardiac disease is frequently reported in farmed animals , and stress has been implicated as a factor for myocardial dysfunction in commercial fish rearing . DB00741 is a major stress hormone in teleosts , and this hormone has adverse effects on the myocardium . Strains of rainbow trout ( Oncorhynchus mykiss ) selected for divergent post-stress cortisol levels [ high responsive ( HR ) and low responsive ( LR ) ] have been established as a comparative model to examine how fish with contrasting stress-coping styles differ in their physiological and behavioral profiles . We show that the mean cardiosomatic index ( CSI ) of adult HR fish was 34 % higher than in LR fish , mainly because of hypertrophy of the compact myocardium . To characterize the hypertrophy as physiological or pathological , we investigated specific cardiac markers at the transcriptional level . HR hearts had higher mRNA levels of cortisol receptors ( MR , GR1 and GR2 ) , increased P53805 levels [ suggesting enhanced pro-hypertrophic nuclear factor of activated T-cell ( NFAT ) signaling ] and increased P15692 gene expression ( reflecting increased angiogenesis ) . Elevated collagen ( Col1a2 ) expression and deposition in HR hearts supported enhanced fibrosis , whereas the heart failure markers P01160 and DB04899 were not upregulated in HR hearts . To confirm our results outside the selection model , we investigated the effect of acute confinement stress in wild-type European brown trout , Salmo trutta . A positive correlation between post-stress cortisol levels and CSI was observed , supporting an association between enhanced cortisol response and myocardial remodeling . In conclusion , post-stress cortisol production correlates with myocardial remodeling , and coincides with several indicators of heart pathology , well-known from mammalian cardiology . DB01645 potentiates the P01160 effect on a K(+)-conductance in P29320 -293 cells . P29320 -293 cells are known to reflect many features of the late distal tubule . Furthermore , they have the ability to release urodilatin , the structural analog to P01160 . RT-PCR was performed to test for the expression of natriuretic peptide receptors . While the mRNA for the human P01160 receptor ( P16066 , P16066 ) could be amplified , the P09543 -specific receptor P20594 ( P20594 ) and the receptor specific for guanylins , P25092 , could not be detected . In patch clamp experiments the effects of P01160 ( 10 nM ) on membrane voltage ( V(m) ) were monitored and P29320 -293 cells depolarized by 2.3 +/- 0.5 mV ( n=14 ) . In the presence of the P01133 receptor blocker genistein ( 10 microM ) the effect of P01160 was increased by 65 % to 3.9 +/- 0.8 mV ( n=14 ) . After removal of genistein the P01160 -mediated depolarization further increased by 147 % to 5.7 +/- 1.0 mV ( n=14 ) . P01160 given repetitively without genistein had no increasing depolarizing effect in P29320 -293 cells with time . The P01160 effect could be fully blocked by 1 mM Ba(2+) and by 1 microM of the specific PKG inhibitor KT5823 indicating that P01160 inhibits a K(+)-conductance via a cGMP-dependent protein kinase . DB01645 itself hyperpolarized the membrane voltage of P29320 -293 cells by -3.9 +/- 0.6 mV ( n=11 ) and this effect could also be fully blocked by Ba(2+) ( -0.3 +/- 0.1 mV , n=5 ) , indicating that genistein activates a K(+)-conductance which contributes significantly to the membrane potential of P29320 -293 cells . ω-3 fatty acid eicosapentaenoic acid attenuates P25189 +-induced neurodegeneration in fully differentiated human SH-SY5Y and primary mesencephalic cells . DB00159 ( EPA ) , a neuroactive omega-3 fatty acid , has been demonstrated to exert neuroprotective effects in experimental models of Parkinson 's disease ( PD ) , but the cellular mechanisms of protection are unknown . Here , we studied the effects of EPA in fully differentiated human SH-SY5Y cells and primary mesencephalic neurons treated with P25189 (+) . In both in-vitro models of PD , EPA attenuated an P25189 (+) -induced reduction in cell viability . EPA also prevented the presence of electron-dense cytoplasmic inclusions in SH-SY5Y cells . Then , possible mechanisms of the neuroprotection were studied . In primary neurons , EPA attenuated an P25189 (+) -induced increase in Tyrosine-related kinase B ( TrkB ) receptors . In SH-SY5Y cells , EPA down-regulated reactive oxygen species and nitric oxide . This antioxidant effect of EPA may have been mediated by its inhibition of neuronal NADPH oxidase and cyclo-oxygenase-2 ( P35354 ) , as P25189 (+) increased the expression of these enzymes . Furthermore , EPA prevented an increase in cytosolic phospholipase A2 ( P47712 ) , an enzyme linked with P35354 in the potentially pro-inflammatory arachidonic acid cascade . Lastly , EPA attenuated an increase in the bax:bcl-2 ratio , and cytochrome c release . However , EPA did not prevent mitochondrial enlargement or a decrease in mitochondrial membrane potential . This study demonstrated cellular mechanisms by which EPA provided neuroprotective effects in experimental PD . [(11)C]5-HTP and microPET are not suitable for pharmacodynamic studies in the rodent brain . The PET tracer [(11)C]5-hydroxytryptophan ( [(11)C]5-HTP ) , which is converted to [(11)C]5-hydroxytryptamine ( [(11)C]5-HT ) by aromatic amino acid decarboxylase ( P20711 ) , is thought to measure 5-HT synthesis rates . But can we measure these synthesis rates by kinetic modeling of [(11)C]5-HTP in rat ? Male rats were scanned with [(11)C]5-HTP ( 60 minutes ) after different treatments . Scans included arterial blood sampling and metabolite analysis . 5-HT synthesis rates were calculated by a two-tissue compartment model ( 2TCM ) with irreversible tracer trapping or Patlak analysis . DB00190 ( inhibitor peripheral P20711 ) dose-dependently increased [(11)C]5-HTP brain uptake , but did not influence 2TCM parameters . Therefore , 10 mg/kg carbidopa was applied in all subsequent study groups . These groups included treatment with NSD 1015 ( general P20711 inhibitor ) or p-chlorophenylalanine ( PCPA , inhibitor of tryptophan hydroxylase , P17752 ) . In addition , the effect of a low-tryptophan ( DB00150 ) diet was investigated . NSD 1015 or DB00150 depletion did not affect any model parameters , but PCPA reduced [(11)C]5-HTP uptake , and the k3 . This was unexpected as NSD 1015 directly inhibits the enzyme converting [(11)C]5-HTP to [(11)C]5-HT , suggesting that trapping of radioactivity does not distinguish between parent tracer and its metabolites . As different results have been acquired in monkeys and humans , [(11)C]5-HTP-PET may be suitable for measuring 5-HT synthesis in primates , but not in rodents . Phosphodiesterase-4 influences the PKA phosphorylation status and membrane translocation of G-protein receptor kinase 2 ( P25098 ) in P29320 -293beta2 cells and cardiac myocytes . Membrane-recruitment of P25098 ( G-protein receptor kinase 2 ) provides a fundamental step in the desensitization process controlling GPCRs ( G-protein-coupled receptors ) , such as the beta2AR ( beta2-adrenergic receptor ) . In the present paper , we show that challenge of P29320 -293beta2 [ human embryonic kidney cells stably overexpressing the FLAG-tagged beta2AR-GFP ( green fluorescent protein ) ] cells with the beta-adrenoceptor agonist , isoprenaline , causes P25098 to become phosphorylated by PKA ( DB02527 -dependent protein kinase ) . This action is facilitated when DB02527 -specific DB05876 ( phosphodiesterase-4 ) activity is selectively inactivated , either chemically with rolipram or by siRNA ( small interfering RNA ) -mediated knockdown of Q07343 and Q08499 . DB05876 -selective inhibition by rolipram facilitates the isoprenaline-induced membrane translocation of P25098 , phosphorylation of the beta2AR by P25098 , membrane translocation of beta-arrestin and internalization of beta2ARs . DB05876 -selective inhibition also enhances the ability of isoprenaline to trigger the PKA phosphorylation of P25098 in cardiac myocytes . In the absence of isoprenaline , rolipram-induced inhibition of DB05876 activity in P29320 -293beta2 cells acts to stimulate PKA phosphorylation of P25098 , with consequential effects on P25098 membrane recruitment and P25098 -mediated phosphorylation of the beta2AR . We propose that a key role for DB05876 enzymes is : ( i ) to gate the action of PKA on P25098 , influencing the rate of P25098 phosphorylation of the beta2AR and consequential recruitment of beta-arrestin subsequent to beta-adrenoceptor agonist challenge , and ( ii ) to protect P25098 from inappropriate membrane recruitment in unstimulated cells through its phosphorylation by PKA in response to fluctuations in basal levels of DB02527 . P19099 inhibitors in cardiovascular and renal diseases . DB04630 is involved in various cardiovascular pathologies , including hypertension , heart failure , atherosclerosis and fibrosis . P08235 ( MR ) -dependent and -independent , genomic and non-genomic processes mediate its complex effects . Spironolactone and eplerenone , both MR antagonists , are the only commercially available compounds targeting directly the actions of aldosterone . However , due to the poor selectivity ( spironolactone ) , low potency ( eplerenone ) and the fact that only MR-dependent effects of aldosterone can be inhibited , these drugs have limited clinical use . An attractive approach to abolish potentially all of aldosterone-mediated pathologies is the inhibition of aldosterone synthase . This review summarizes current knowledge on the complex effects mediated by aldosterone , potential advantages and disadvantages of aldosterone inhibition and novel directions in the development of aldosterone synthase inhibitors . Gastroduodenal tolerability of lumiracoxib vs placebo and naproxen : a pilot endoscopic study in healthy male subjects . BACKGROUND : DB01283 ( DB01283 ) is a cyclooxygenase-2 ( P35354 ) selective inhibitor . AIM : To compare the gastroduodenal tolerability of lumiracoxib with placebo and naproxen in a randomized , parallel-group , double-blind study . METHODS : : Sixty-five healthy male subjects were randomized to receive 8 days ' dosing with lumiracoxib 200 mg twice daily ( b.d. ) ( n = 21 ) , placebo ( n = 22 ) or naproxen 500 mg b.d . ( n = 22 ) . Endoscopic evaluations of gastric and duodenal mucosae were conducted at baseline and after 8 days ' dosing . Serum was assayed for ex-vivo concentrations of thromboxane B2 ( TxB2 ) to determine cyclooxygenase-1 ( P23219 ) inhibitory activity . RESULTS : Sixty subjects ( 20 per group ) completed the study . No gastroduodenal erosions were observed in subjects receiving lumiracoxib . Thirteen subjects receiving naproxen developed duodenal erosions . At the gastric site , one subject in each of the naproxen and placebo groups had erosions ; one subject receiving naproxen also developed a small asymptomatic gastric ulcer . Gastrointestinal adverse events accounted for 42.3 % of all adverse events , occurring in 3/21 , 4/22 and 6/22 of the lumiracoxib , placebo and naproxen groups , respectively . TxB2 levels were similar for patients receiving placebo or lumiracoxib , but were reduced by > 95 % in patients receiving naproxen , compared with placebo . CONCLUSIONS : Multiple doses of lumiracoxib resulted in gastroduodenal tolerability similar to placebo and superior to naproxen . Paullones are potent inhibitors of glycogen synthase kinase-3beta and cyclin-dependent kinase 5/p25 . Paullones constitute a new family of benzazepinones with promising antitumoral properties . They were recently described as potent , DB00171 -competitive , inhibitors of the cell cycle regulating cyclin-dependent kinases ( CDKs ) . We here report that paullones also act as very potent inhibitors of glycogen synthase kinase-3beta ( GSK-3beta ) ( IC50 : 4-80 nM ) and the neuronal Q00535 /p25 ( IC50 : 20-200 nM ) . These two enzymes are responsible for most of the hyperphosphorylation of the microtubule-binding protein tau , a feature observed in the brains of patients with Alzheimer 's disease and other neurodegenerative ' taupathies ' . DB04014 , the most active paullone , was demonstrated to act by competing with DB00171 for binding to GSK-3beta . DB04014 inhibits the phosphorylation of tau in vivo at sites which are typically phosphorylated by GSK-3beta in Alzheimer 's disease . DB04014 also inhibits the Q00535 /p25-dependent phosphorylation of Q9UD71 in mouse striatum slices in vitro . This dual specificity of paullones may turn these compounds into very useful tools for the study and possibly treatment of neurodegenerative and proliferative disorders . Decreased fasting blood glucose is associated with impaired hepatic glucose production in thyroid-stimulating hormone receptor knockout mice . Our previous study reported that thyroid-stimulating hormone ( DB00024 ) promotes cholesterol synthesis via the cyclic adenosine monophosphate/protein kinase A/ DB02527 regulatory element-binding protein ( DB02527 /PKA/CREB ) pathway after binding to DB00024 receptors ( P16473 ) in the liver . The hepatic DB02527 /PKA/CREB pathway also plays an important role in maintaining fasting glucose homeostasis . These findings implied a possible role for DB00024 in hepatic glucose metabolism . In this study , we used DB00024 receptor knockout mice ( Tshr-ko mice ) to clarify the effect of Tshr deletion on hepatic glucose metabolism , and investigated whether the effects of DB00024 directly regulate hepatic gluconeogenesis in HepG2 cells . Tshr-ko mice exhibited decreased fasting blood glucose levels , increased insulin sensitivity but normal level of fasting plasma insulin . Tshr deletion impaired hepatic glucose production by down-regulating the expression of glucose-6-phosphatase ( G6P ) and phosphoenolpyruvate pyruvate carboxylase ( PEPCK ) mRNA , two rate-limiting enzymes in hepatic gluconeogenesis , and enhancing the abundance of hepatic glucokinase ( GK ) , the first enzyme regulating glycogen synthesis . Moreover , Tshr deletion inhibited the protein expression of hepatic phospho-CREB and increased the protein expression of hepatic phospho-AMP-activated protein kinase ( p-AMPK ) , two up-stream regulators of PEPCK and G6P mRNA . In HepG2 cells , DB00024 increased the expression of G6P and PEPCK at mRNA level . These results indicated the simulative effects of DB00024 on hepatic glucose production in vivo and in vitro , suggesting a novel role for DB00024 in hepatic glucose metabolism . Effect of pregnancy on DB05876 DB02527 -specific phosphodiesterase messenger ribonucleic acid expression in human myometrium . In light of the important role of the second messengers DB02527 and cGMP in the mechanism of relaxation in the human myometrium , specific regulation of the phosphodiesterase ( PDE ) enzymatic system responsible for cyclic nucleotide inactivation is essential . We previously identified in the human myometrium DB05876 DB02527 -specific PDE as by far the most abundant isoform . Here we have studied the expression patterns of mRNAs for the four cloned human DB05876 genes in the myometria of pregnant and non-pregnant women . Concurrent expression of the P27815 , 4B , 4C and 4D genes is demonstrated . We found that the Q08499 transcripts are the most prominently expressed . P27815 and Q07343 mRNAs also are markedly abundant , whereas lower expression is observed for Q08493 mRNAs . Interestingly , we showed that transcripts of PDE4B2 are more abundant in the myometria of pregnant women than in non-pregnant women , whereas no difference between the two tissues was detected for P27815 , 4C and 4D mRNAs . Cultured human myometrial cells , which present a high level of DB05876 activity and express the four DB05876 mRNA subtypes , provide us with an appropriate model to further evaluate whether the level of expression of the PDE 4B2 mRNA subtype is under hormonal regulation . P20711 inhibition does not influence the diuretic and natriuretic response to exogenous alpha-atrial natriuretic peptide in man . The role of dopamine synthesis in the renal actions of human alpha-atrial natriuretic peptide ( alpha P01160 ) was investigated in six dehydrated volunteers using the P20711 inhibitor carbidopa . Each subject received oral placebo or carbidopa ( 100 mg ) followed by an infusion of alpha P01160 10 pmol.kg-1.min-1 for 1 h . The responses to placebo alone and to carbidopa alone were investigated on separate occasions. alpha P01160 produced a similar increase in plasma immunoreactive alpha P01160 whether placebo or carbidopa pretreatment had been given . Urinary dopamine excretion was increased by alpha P01160 . DB00190 pretreatment substantially attenuated this increase without affecting the natriuretic or water-diuretic response to alpha P01160 . DB00190 also failed to alter the change in filtration fraction produced by alpha P01160 . The results suggest that increased synthesis of intrarenal dopamine is not required for the renal effects of alpha P01160 in man . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . DB01269 : a new frontier of target therapy for the treatment of metastatic colorectal cancer . DB01269 is a fully human monoclonal IgG(2) antibody targeting the P01133 receptor ( P00533 ) . This agent represents a new class of drug owing to its fully human nature , and no need for premedication and loading dose . DB01269 selectively binds to P00533 , blocking the extracellular domain of the receptor , and has not been associated with the formation of any antibodies directed against it . The drug is indicated as monotherapy for the treatment of patients with P00533 -expressing metastatic colorectal carcinoma with non-mutated ( wild-type ) P01116 after failure of fluoropyrimidine- , oxaliplatin- and irinotecan-containing chemotherapy regimens . The safety profile is favorable and is generally well tolerated ; the most common toxicities are skin rashes and diarrhea . Therefore , panitumumab 's hypersensitivity reaction rate is lower when compared with a chimeric monoclonal antibody such as cetuximab . DB01269 increases the clinician 's repertoire of agents to treat metastatic colorectal carcinoma . The available clinical data are the most promising for a single-agent anti- P00533 monoclonal antibody in this disease at the present time . These new data open different clinical scenarios in metastatic colorectal carcinoma patients and encourage clinicians and basic researchers to investigate new therapeutic approaches for this patient subset . Altered regulation of renal nitric oxide , atrial natriuretic peptide and cyclooxygenase systems in aldosterone escape in rats . The present study was aimed to determine whether there is an altered role of local nitric oxide ( NO ) , atrial natriuretic peptide ( P01160 ) and cyclooxygenase ( P36551 ) systems in the kidney in association with the aldosterone escape . Male Sprague-Dawley rats were used . DB04630 ( 200 microg/day ) was infused through entire time course . The control group was kept on a low sodium diet ( 0.02 mEq/day ) , and the experimental group was supplied with a higher sodium diet ( 2. /day ) . Four days after beginning the regimen , the kidneys were taken . The protein expression of NO synthase ( NOS ) and P36551 isoforms was determined by semiquantitative immunoblotting . The mRNA expression of components of P01160 system was determined by real-time polymerase chain reaction . The activities of soluble and particulate guanylyl cyclases were determined by the amount of cGMP generated in responses to sodium nitroprusside and P01160 , respectively . There developed aldosterone escape in the experimental group . Accordingly , the renal content and the urinary excretion of NO increased . The expression of P29475 was increased in the inner medulla . Neither the expression of P29474 nor that of P35228 was changed . The expression and the catalytic activity of soluble guanylyl cyclase remained unaltered . The mRNA expression of P01160 was increased . Neither the expression of P16066 or P17342 nor the activity of particulate guanylyl cyclase was altered in the papilla . The protein expression of P35354 was increased in the inner medulla , while that of P23219 remained unchanged . In conclusion , the upregulation of P29475 , P01160 , and P35354 may be causally related with the aldosterone escape . Cloning and functional characterization of a testicular DB00024 receptor cDNA from the African catfish ( Clarias gariepinus ) . A cDNA encoding a putative thyroid-stimulating hormone receptor ( cfTSH-R ) was cloned from the testis of the African catfish ( Clarias gariepinus ) . The cfTSH-R showed the highest amino acid sequence identity with the DB00024 -Rs of other fish species . In addition , an insertion of approximately 50 amino acids , specific for the P16473 subfamily , was also present in the carboxy terminus of the amino-terminal extracellular domain of the cfTSH-R . Next to the testis and thyroid follicles , abundant cfTSH-R expression was detected in cerebellum , brain , ovary , seminal vesicles and pituitary , while weaker expression was found in muscle , stomach , intestine , head-kidney , liver , kidney and heart . P29320 -T 293 cells , transiently expressing the cfTSH-R , significantly increased intracellular DB02527 levels in response to human DB00024 . Catfish LH , human choriogonadotropin and human DB00094 were also able to induce this cfTSH-R-mediated response , although with considerably lower efficiency than human DB00024 . These results indicated that a functional cfTSH-R had been cloned from the testis of African catfish .

MH_train_1326

MH_train_1326

MH_train_1326

interacts_with DB00379?

multiple_choice

Gating properties of Q14524 mutations and the response to mexiletine in long-QT syndrome type 3 patients . BACKGROUND : DB00379 ( Mex ) has been proposed as a gene-specific therapy for patients with long-QT syndrome type 3 ( LQT3 ) caused by mutations in the cardiac sodium channel gene ( Q14524 ) . The degree of QT shortening and the protection from arrhythmias vary among patients harboring different mutations . We tested whether the clinical response to Mex in LQT3 could be predicted by the biophysical properties of the different mutations . METHODS AND RESULTS : We identified 4 Q14524 mutations in 5 symptomatic LQT3 patients with different responses to Mex ( 6 to 8 mg . kg(-1) . d(-1) ) . We classified the mutations as sensitive to Mex ( P1332L , R1626P ; >/= 10 % of QTc shortening and QTc < 500 ms or no arrhythmias ) or insensitive to Mex ( S941N , M1652R ; negligible or no QTc shortening and sudden death ) . We measured Na(+) current from P29320 293 cells transfected with wild-type ( WT ) or mutant Nav1.5 . All mutations showed impaired inactivation of Na(+) current , but the mutations identified in patient responders to Mex ( P1332L , R1626P ) showed a hyperpolarizing shift of V(1/2) of steady-state inactivation . Furthermore , Mex produced use-dependent block with the order R1626P=P1332L > S941N=WT > M1652R , suggesting that Mex-sensitive mutants present prolonged recovery from Mex block . CONCLUSIONS : We propose that voltage dependence of channel availability and shifts of V(1/2) of steady-state inactivation correlate with the clinical response observed in LQT3 patients . This supports the view that the response to Mex is mutation specific and that in vitro testing may help to predict the response to therapy in LQT3 . Interaction between the insulin receptor and Grb14 : a dynamic study in living cells using BRET . Grb14 is a molecular adaptor that binds to the activated insulin receptor ( IR ) and negatively regulates insulin signaling . We have studied the dynamics of interaction of the IR with Grb14 , in real time , in living P29320 cells , using bioluminescence resonance energy transfer ( BRET ) . P01308 rapidly and dose-dependently stimulated this interaction . Removing insulin from the incubation medium only resulted in a modest decrease in BRET signal , indicating that the interaction between the IR and Grb14 can remain long after insulin stimulus has disappeared . BRET saturation experiments indicated that insulin markedly increases the affinity between IR and Grb14 , resulting in recruitment of the adaptor to the activated IR . In addition , using both BRET and co-immunoprecipitation experiments , we demonstrated that insulin induced the dimerization of Grb14 , most likely as a result of simultaneous binding of two Grb14 molecules on the activated IR . We also investigated the relationships between IR , Grb14 and the protein tyrosine phosphatase P18031 . We observed that insulin-induced BRET between the IR and P18031 was markedly reduced by Grb14 , suggesting that Grb14 regulated this interaction in living cells . Using site-specific antibodies against phosphorylated tyrosines of the insulin receptor , we showed that Grb14 protected the three tyrosines of the kinase loop from dephosphorylation by P18031 , while favouring dephosphorylation of tyrosine 972 . This resulted in decreased P35568 binding to the IR and decreased activation of the P29323 pathway . Our work suggests that Grb14 may regulate signalling through the insulin receptor by controlling its tyrosine-dephosphorylation in a site-specific manner . Is abiraterone acetate well tolerated and effective in the treatment of castration-resistant prostate cancer ? This Practice Point commentary discusses the findings of the first phase I trial to evaluate abiraterone acetate ( an inhibitor of the androgen-regulating enzyme P05093 ) in the treatment of castration-resistant prostate cancer . This open-label , dose-escalation study by Attard et al. showed that abiraterone was well tolerated but often induced a syndrome of secondary mineralocorticoid excess that improved with eplerenone ( a mineralocorticoid receptor antagonist ) . DB05812 is a potent suppressor of adrenal androgen synthesis , and produced lasting prostate-specific antigen responses in approximately half of the patients . A few patients had partial regression of distant metastases . Although promising , these results should be interpreted with caution owing to the small sample size and because the study was not primarily designed to examine drug efficacy . Multi-institutional , prospective trials should provide additional information on the tolerability and activity of this compound and further define the population most likely to benefit from this endocrine approach . Novel treatments of GERD : focus on the lower esophageal sphincter . Up to 50 % of patients with gastroesophageal reflux disease ( GERD ) still suffer from GERD symptoms despite proton pump inhibitor ( PPI ) therapy , indicating a need for new treatments . The lower esophageal sphincter ( LES ) plays a crucial role in maintaining the mechanical barrier necessary for prevention of gastric reflux . Transient LES relaxation ( TLESR ) is an important factor behind the occurrence of reflux , and preclinical studies have identified a number of targets for pharmacologic modification of TLESR . However , only gamma-aminobutyric acid ( GABA ) type B receptor ( GABA(B) ) agonists and metabotropic glutamate receptor 5 ( P41594 ) modulators have shown positive proof of concept in the clinical setting . The P41594 negative allosteric modulator DB05070 improved symptoms in GERD patients , but was associated with central side effects such as dizziness . DB00181 , a GABA(B) receptor agonist , reduces the incidence of TLESR and improves GERD symptoms in both adult and pediatric GERD patients . However , the utility of baclofen is similarly limited by poor tolerability and recent research has focused on the development of GABA(B) receptor agonists with improved tolerability . DB05031 , a prodrug of R-baclofen , reduced the number of reflux episodes in a dose-ranging study and was similarly tolerated to placebo . AZD3355 and AZD9343 are GABA(B) receptor agonists with limited central nervous system activity that have been shown in preclinical studies to reduce the incidence of TLESR and decrease esophageal acid exposure ; data from clinical studies of these agents in GERD patients are awaited with interest . Agents that target TLESR activity may therefore offer a promising new add-on treatment for patients who suffer from GERD symptoms despite PPI therapy . Activated human neutrophils rapidly release the chemotactically active D2D3 form of the urokinase-type plasminogen activator receptor ( Q03405 /CD87 ) . The urokinase-type plasminogen activator receptor ( Q03405 /CD87 ) exists both in cell-bound and soluble forms . Neutrophils contain extensive intracellular pools of Q03405 that are translocated to the plasma membrane upon activation . In the present study , we investigated the ability of human neutrophils to shed Q03405 from cell surface following activation and addressed the possible involvement of the released receptor in the inflammatory response . We first observed that the spontaneous release of suPAR by resting neutrophils was strongly and rapidly ( within minutes ) enhanced by calcium ionophore ionomycin and to a lesser extent when cells were primed with P01375 and then stimulated with fMLP or P10145 . We demonstrated that suPAR is produced by resting and activated neutrophils predominantly as a truncated form devoid of N-terminal D1 domain ( D2D3 form ) that lacks P06744 anchor . Migration of formyl peptide receptor-like 1 ( P25090 ) -transfected human embryonic kidney ( P29320 ) 293 cells toward the supernatants harvested from activated neutrophils was significantly diminished when D2D3 form of suPAR was immunodepleted from the supernatants . We conclude that activated neutrophils release the chemotactically active D2D3 form of suPAR that acts as a ligand of P25090 . Interestingly , we present evidence that P80108 ( P80108 ) that has previously been shown to shed Q03405 in cancer cells is not involved in suPAR release from human neutrophils . We suggest that production of the chemotactically active D2D3 form of suPAR by activated human neutrophils in vivo could contribute to the recruitment of monocytes and other formyl peptide receptors-expressing cells to the sites of acute inflammation where neutrophil accumulation and activation occur . Encapsulation of viral vectors for gene therapy applications . In gene therapy , a number of viruses are currently being used as vectors to provide transient expression of therapeutic proteins . A drawback of using free virus is that it gives a potent immune response , which reduces gene transfer and limits re-administration . An alternative delivery system is to encapsulate the virus in poly(lactide-co-glycolide) ( P00747 ) microspheres prior to administration . A recombinant adenovirus ( Ad ) expressing green fluorescent protein ( GFP ) was used to test the transduction efficiency of Ad encapsulated in microspheres on target cells . The number of infected cells that expressed GFP was measured by flow cytometry . It was demonstrated that encapsulated viral vectors could successfully transduce target cells with encapsulation efficiencies up to 23 % and that the level of transduction could be controlled by varying both the quantity of microspheres and the amount of Ad in the microspheres . High transduction efficiencies and its recognized biocompatibility make P00747 -encapsulated Ad an attractive alternative to the use of free virus in gene therapy applications . The infectivity of Ad was found to be significantly influenced by the processing conditions and changes in environmental factors . Free Ad and encapsulated Ad were able to infect both E1 complimenting cells ( P29320 293 ) and non-complimenting cells ( A549 ) , with the viral expression in P29320 293 cells being 2.1 times greater than for A549 cells . Detection and quantification of cimicoxib , a novel P35354 inhibitor , in canine plasma by HPLC with spectrofluorimetric detection : development and validation of a new methodology . DB05095 ( CX ) is a selective P35354 inhibitor recently launched on the veterinary market . No analytical method to detect CX in biological samples has been published to date . The chromatographic separation was performed with a Kinetex C18 analytical column ( 100 mm × 4.6 mm , 2.6 μm particle size ) at 25 °C . The mobile phase consisted of acetonitrile:buffer ( 10 mM AcONH4 , pH 4.5 ) ( 35:65 , v/v ) at a flow rate of 1 mL/min . Excitation and emission wavelengths were 268 and 430 nm , respectively . The extraction used 500 μL of plasma added to 100 μL of IS ( 5 μg/mL ) and 100 μL of 10 % CF3COOH , extracted with 600 μL of C2H2:Et2O ( 3:7 , v/v ) . The organic phase was evaporated and reconstituted with 200 μL of mobile phase . The CX recovery ranged from 74.5 % to 82.6 % . The limit of quantification was 25 ng mL(-1) . The chromatographic runs were specific with no interfering peaks at the retention times of the analytes , as confirmed by HPLC-mass spectrometry experiments . The other validation parameters were in agreement with the international guidelines . The method was successfully tested on two dogs treated at two dose rates . It facilitated tracking of the plasma concentration for 24h and calculation of the main pharmacokinetic parameters . In conclusion , this method ( extraction , separation and applied techniques ) is simple , effective and specific . This is the first time that a method for the quantification of CX in plasma has been reported . This technique may have applications for further pharmacokinetic studies . Substituent effects of N-(1,3-diphenyl-1H-pyrazol-5-yl)benzamides on positive allosteric modulation of the metabotropic glutamate-5 receptor in rat cortical astrocytes . CDPPB [ 3-cyano-N-(1,3-diphenyl-1H-pyrazol-5-yl)benzamide ] was recently described as the first centrally active , positive allosteric modulator of rat and human metabotropic glutamate receptor ( mGluR ) P41594 subtype . We explored the structural requirements for potentiation of glutamate-induced calcium release in naturally expressed P41594 in cultured rat astrocytes and increasing affinity for the allosteric antagonist binding site by evaluating 50 analogues of CDPPB . In the fluorometric calcium assay , CDPPB exhibited an EC50 value of 77 +/- 15 nM in potentiating P41594 -mediated responses in cortical astrocytes and a Ki value of 3760 +/- 430 nM in displacing [3H]methoxyPEPy binding in membranes of cultured P29320 -293 cells expressing rat P41594 . The structure-activity relationships showed that electronegative aromatic substituents in the para-position of the benzamide moiety of CDPPB increase potency . Both binding and functional activities were further increased with a halogen atom in the ortho-position of the 1-phenyl ring . These effects of substitution do not match those of either aromatic ring of MPEP [ 2-methyl-6-(phenylethynyl)pyridine ] for the antagonist allosteric binding site . Combination of the optimal substituents and aromatic positions resulted in 4-nitro-N-(1-(2-fluorophenyl)-3-phenyl-1H-pyrazol-5-yl)benzamide ( VU-1545 ) showing Ki = 156 +/- 29 nM and EC50 = 9.6 +/- 1.9 nM in the binding and functional assays , respectively . [ The chemical structure and pharmacological properties of a novel isoxazolidinedione insulin sensitizer , DB04971 ] . DB04971 is an isoxazolidine-3,5-dione derivative . This drug activates both Q07869 gamma and Q07869 alpha , and shows not only a hypoglycemic effect but also a stronger triglyceride-lowering effect than the thiazolidine-2,4-diones . DB04971 improved both the impaired insulin-stimulated autophosphorylation levels of Zucker fatty rats and impaired insulin-induced P14672 translocation to the plasma membrane as well as insulin-induced glucose uptake in high fat diet rats , indicating that DB04971 enhances insulin signaling and reduces insulin resistance . Furthermore , DB04971 prevented several diabetic complications , such as cataract , nephropathy , and neuropathy in Zucker diabetic fatty rats . As a non-thiazolidinedione insulin sensitizer , DB04971 has been the first to start clinical trials and is currently undergoing evaluation in clinical studies for diabetic patients . Technique appraisement of comparative proteomics and screening of differentiation-related protein in gastric carcinoma . BACKGROUND/AIMS : Different differentiations of cancer have resulted in its unique biological characteristics . We screen and appraise differentially expressed proteins in different differentiated gastric adenocarcinoma with comparative proteomics technology in order to find regulatory factors of tumor differentiation related and finally reach the purpose of tumor differentiation reversal . METHODOLOGY : With two-dimensional fluorescence difference gel electrophoresis ( 2-D DIGE ) and liquid chromatography in conjunction with tandem mass spectrometry ( LC–MS/MS ) , the differentially expressed proteins from 8 patients with different differentiated gastric adenocarcinoma were identified and some factors identified were verified with application of QPCR and Western blot techniques . RESULTS : Significant differences in 35 protein spots were found and 48 kinds of proteins were identified . Other than structural proteins and non-specific protein , six possible proteins associated with tumor differentiation were determined - the serine protease inhibitor B1 ( serine protease inhibitor , clade B , member 1 , P30740 ) , calcium-phospholipid binding protein III ( annexin A3 ) , transcription factor Nm23-H1 , adenine phosphoribosyl-transferase enzyme P07741 ( DB00173 Phosphoribosyltransferase in APO and AMP ) , glutathione S-transferase P1-1 ( Q86UG4 -π-1 ) , antimicrobial peptides P81605 -lL . The identified P30740 , annexin A3 , Nm23-H1 and P07741 were verified , confirming the expression of these factors was in line with proteomics identification . CONCLUSIONS : Protein expression in different differentiated gastric adenocarcinoma was varied . Clathrin-dependent internalization of the angiotensin II AT₁A receptor links receptor internalization to P35354 protein expression in rat aortic vascular smooth muscle cells . The major effects of Angiotensin II ( AngII ) in vascular tissue are mediated by AngII AT1A receptor activation . Certain effects initiated by AT1A receptor activation require receptor internalization . In rat aortic vascular smooth muscle cells ( RASMC ) , AngII stimulates cyclooxygenase 2 protein expression . We have previously shown this is mediated by β-arrestin-dependent receptor internalization and NF-κB activation . In this study , a specific inhibitor of clathrin-mediated endocytosis ( CME ) , pitstop-2 , was used to test the hypothesis that clathrin-dependent internalization of activated AT1A receptor mediates NF-κB activation and subsequent cyclooxygenase 2 expression . Radioligand binding assays , real time qt-PCR and immunoblotting were used to document the effects of pitstop-2 on AngII binding and signaling in RASMC . Laser scanning confocal microscopy ( LSCM ) was used to image pitstop-2׳s effects on AT1 receptor/GFP internalization in P29320 -293 cells and p65 NF-κB nuclear localization in RASMC . Pitstop-2 significantly inhibited internalization of AT1A receptor ( 44.7 % ± 3.1 % Control vs. 13.2 % ± 8.3 % Pitstop-2 ; n=3 ) as determined by radioligand binding studies in RASMC . Studies utilizing AT1A receptor/GFP expressed in P29320 293 cells and LSCM confirmed these findings . Pitstop-2 significantly inhibited AngII-induced p65 NF-κB phosphorylation and nuclear localization , P35354 message and protein expression in RASMC without altering activation of Q8NFH3 /44 P29323 or TNFα signaling . Pitstop-2 , a specific inhibitor of clathrin-mediated endocytosis , confirms that internalization of activated AT1A receptor mediates AngII activation of cyclooxygenase 2 expression in RASMC . These data provide support for additional intracellular signaling pathways activated through β-arrestin mediated internalization of G protein-coupled receptors , such as AT1A receptors . [ Synthesis of ( 2'-bromo-4 ' , 5'-dimethoxy-phenyl ) -(2,3-dibromo-4,5-dimethoxy-phenyl)-methane as P18031 inhibitor ] . OBJECTIVE : To synthesize (2'-bromo-4',5'-dimethoxy-phenyl)- ( 2,3- dibromo-4,5-dimethoxy-phenyl ) -methane ( 6 ) as protein tyrosine phosphatase 1B ( P18031 ) inhibitor . METHOD : DB04088 was synthesized by Friedel-Crafts reaction , bromination and decarbonylation and screened inhibitory activity against P18031 by the colorimetric assay . The structure of synthetic intermediates and target product were identified on the basis of spectral analysis . RESULT : DB04088 was synthesized successfully in four steps and evaluated for its P18031 inhibitory activity , the screening result shown that compound 6 displayed good inhibitory activity against P18031 . CONCLUSION : The target compound 6 was synthesized with the overall yield of 20 % , which was a new compound and shown good inhibitory activity against P18031 ( inhibition 40.16 % at 5 mg x L(-1) ) . DB11582 suppresses osteoclastogenesis induced by O14788 and cancer cells through inhibition of inflammatory pathways : a new use for an old drug . BACKGROUND AND PURPOSE : Most patients with cancer die not because of the tumour in the primary site , but because it has spread to other sites . Common tumours , such as breast , multiple myeloma , and prostate tumours , frequently metastasize to the bone . To search for an inhibitor of cancer-induced bone loss , we investigated the effect of thiocolchicoside , a semi-synthetic colchicoside derived from the plant Gloriosa superba and clinically used as a muscle relaxant , on osteoclastogenesis induced by receptor activator of NF-κB ligand ( O14788 ) and tumour cells . EXPERIMENTAL APPROACH : We used RAW 264.7 ( murine macrophage ) cells , a well-established system for osteoclastogenesis , and evaluated the effect of thiocolchicoside on O14788 -induced NF-κB signalling and osteoclastogenesis as well as on osteoclastogenesis induced by tumour cells . KEY RESULTS : DB11582 suppressed osteoclastogenesis induced by O14788 , and by breast cancer and multiple myeloma cells . Inhibition of the NF-κB pathway was responsible for this effect since the colchicoside inhibited O14788 -induced NF-κB activation , activation of IκB kinase ( IKK ) and suppressed inhibitor of NF-κBα ( IκBα ) phosphorylation and degradation , an inhibitor of NF-κB . Furthermore , an inhibitor of the IκBα kinase γ or NF-κB essential modulator , the regulatory component of the IKK complex , demonstrated that the NF-κB signalling pathway is mandatory for osteoclastogenesis induced by O14788 . CONCLUSIONS AND IMPLICATIONS : Together , these data suggest that thiocolchicoside significantly suppressed osteoclastogenesis induced by O14788 and tumour cells via the NF-κB signalling pathway . Thus , thiocolchicoside , a drug that has been used for almost half a century to treat muscle pain , may also be considered as a new treatment for bone loss . β-Arrestin-1 mediates thyrotropin-enhanced osteoblast differentiation . DB00024 ( DB00024 ) activation of the DB00024 receptor ( P16473 ) , a 7-transmembrane-spanning receptor ( 7TMR ) , may have osteoprotective properties by direct effects on bone . P16473 activation by DB00024 phosphorylates protein kinases P31749 , p38α , and P27361 /2 in some cells . We found DB00024 -induced phosphorylation of these kinases in 2 cell lines engineered to express TSHRs , human embryonic kidney P29320 - P16473 cells and human osteoblastic U2OS- P16473 cells . In U2OS- P16473 cells , DB00024 up-regulated pAKT1 ( 7.1±0.5-fold ) , p38α ( 2.9±0.4-fold ) , and pERK1/2 ( 3.1±0.2-fold ) , whereas small molecule P16473 agonist P06681 had no or little effect on pAKT1 ( 1.8±0.08-fold ) , p38α ( 1.2±0.09-fold ) , and pERK1/2 ( 1.6±0.19-fold ) . Furthermore , DB00024 increased expression of osteoblast marker genes P05186 ( 8.2±4.6-fold ) , O14788 ( 21±5.9-fold ) , and osteopontin ( P10451 ; 17±5.3-fold ) , whereas P06681 had little effect ( P05186 , 1.7±0.5-fold ; O14788 , 1.3±0.6-fold ; and P10451 , 2.2±0.7-fold ) . β-Arrestin-1 and -2 can mediate activatory signals by 7TMRs . DB00024 stimulated translocation of β-arrestin-1 and -2 to P16473 , whereas P06681 failed to translocate either β-arrestin . Down-regulation of β-arrestin-1 by siRNA inhibited DB00024 -stimulated phosphorylation of P27361 /2 , p38α , and P31749 , whereas down-regulation of β-arrestin-2 increased phosphorylation of P31749 in both cell types and of P27361 /2 in P29320 - P16473 cells . Knockdown of β-arrestin-1 inhibited DB00024 -stimulated up-regulation of mRNAs for P10451 by 87 ± 1.7 % and O14788 by 73 ± 2.4 % , and P10451 secretion by 74 ± 10 % . We conclude that DB00024 enhances osteoblast differentiation in U2OS cells that is , in part , caused by activatory signals mediated by β-arrestin-1 . Two intronic mutations cause 17-hydroxylase deficiency by disrupting splice acceptor sites : direct demonstration of aberrant splicing and absent enzyme activity by expression of the entire P05093 gene in P29320 -293 cells . To date , only two among 46 mutations in the P05093 gene cause 17-hydroxylase deficiency ( 17OHD ) by disrupting mRNA splice donor sites . We studied two subjects with intronic P05093 mutations : a compound heterozygote for Y329D plus an AG to CG substitution at the 3' end of intron 2 , and a homozygote for a TTTT deletion near the 3' end of intron 3 . We hypothesized that both mutations caused 17OHD by disrupting splice acceptor sites . To prove this mechanism , the entire P05093 genes ( wild type and both mutations ) were amplified , subcloned into pcDNA3 , and expressed in P29320 -293 cells . The mRNA derived from the wild-type P05093 gene was correctly spliced and translated into active enzyme , as shown by the correct sequence in the RT-PCR products and by the 17-hydroxylation of progesterone . In contrast , cells expressing the mutant genes had no 17-hydroxylase activity . The mRNA derived from the AG to CG mutation used the first AG in exon 3 as the splice acceptor site , shifting the reading frame and introducing a stop codon . RNA derived from the TTTT deletion skipped exon 4 entirely , deleting 29 amino acids in-frame . Our data show that these are the first two 17OHD cases resulting from mutations that alter splice acceptor sites . These studies also demonstrate the feasibility of expressing the entire P05093 gene , with simultaneous protein and RNA analysis , as a general methodology for characterizing how intronic P05093 mutations cause 17OHD . Inhibition of the invasion capacity of carcinoma cells by DB05476 , 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 , a novel 3-amidinophenylalanine-based inhibitor of the uPA system , at inhibiting the invasive capacity of carcinoma cells . First , Q03405 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 expression levels correlated positively with invasion capacity , which could be significantly inhibited by DB05476 . 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 . Thus , our results demonstrate the potential of DB05476 in vitro as a promising adjuvant antimetastatic therapy of carcinomas . Immunohistochemical detection of alpha1E voltage-gated Ca(2+) channel isoforms in cerebellum , P01308 -1 cells , and neuroendocrine cells of the digestive system . Polyclonal antibodies were raised against a common and a specific epitope present only in longer alpha1E isoforms of voltage-gated Ca(2+) channels , yielding an " anti-E-com " and an " anti-E-spec " serum , respectively . The specificity of both sera was established by immunocytochemistry and immunoblotting using stably transfected P29320 -293 cells or membrane proteins derived from them . Cells from the insulinoma cell line P01308 -1 , tissue sections from cerebellum , and representative regions of gastrointestinal tract were stained immunocytochemically . P01308 -1 cells expressed an alpha1E splice variant with a longer carboxy terminus , the so-called alpha1Ee isoform . Similarily , in rat cerebellum , which was used as a reference system , the anti-E-spec serum stained somata and dendrites of Purkinje cells . Only faint staining was seen throughout the cerebellar granule cell layer . After prolonged incubation times , neurons of the molecular layer were stained by anti-E-com , suggesting that a shorter alpha1E isoform is expressed at a lower protein density . In human gastrointestinal tract , endocrine cells of the antral mucosa ( stomach ) , small and large intestine , and islets of Langerhans were stained by the anti-E-spec serum . In addition , staining by the anti-E-spec serum was observed in Paneth cells and in the smooth muscle cell layer of the lamina muscularis mucosae . We conclude that the longer alpha1Ee isoform is expressed in neuroendocrine cells of the digestive system and that , in pancreas , alpha1Ee expression is restricted to the neuroendocrine part , the islets of Langerhans. alpha1E therefore appears to be a common voltage-gated Ca(2+) channel linked to neuroendocrine and related systems of the body . Vampire bat plasminogen activator DB04925 -alpha-1 ( desmoteplase ) : a thrombolytic drug optimized by natural selection . P00747 activators are enzymes found in all vertebrate species investigated so far . Their physiological function is the generation of localized proteolysis in the context of tissue remodeling , wound healing and neuronal plasticity . The common vampire bat ( Desmodus rotundus ) is a New World species that feeds exclusively on blood . Its saliva contains highly potent plasminogen activators , specialized in rapid lysis of fresh blood clots . Biochemical and pharmacological evidence indicates that these plasminogen activators represent a new class of thrombolytics with pharmacological and toxicological properties superior to human tissue-type plasminogen activator , the clot dissolving agent now most frequently used in medicine . A form of the enzyme produced by recombinant DNA technology is currently employed to test this hypothesis in clinical studies . Rat peroxisome proliferator-activated receptors and brown adipose tissue function during cold acclimatization . Brown adipose tissue ( Q14032 ) hyperplasia is a fundamental physiological response to cold ; it involves an acute phase of mitotic cell growth followed by a prolonged differentiation phase . Peroxisome proliferator-activated receptors ( PPARs ) are key regulators of fatty acid metabolism and adipocyte differentiation and may therefore mediate important metabolic changes during non-shivering thermogenesis . In the present study we have investigated Q07869 mRNA expression in relation to peroxisome proliferation in rat Q14032 during cold acclimatization . By immunoelectron microscopy we show that the number of peroxisomes per cytoplasmic volume and acyl- DB01992 oxidase immunolabeling density remained constant ( thus increasing in parallel with tissue mass and cell number ) during the initial proliferative phase and the acute thermogenic response but increased after 14 days of cold exposure , correlating with terminal differentiation of Q14032 . A pronounced decrease in Q14032 PPARalpha and PPARgamma mRNA levels was found within hours of exposure to cold , which was reversed after 14 days , suggesting a role for either or both of these subtypes in the proliferation and induction of peroxisomes and peroxisomal beta-oxidation enzymes . In contrast , PPARdelta mRNA levels increased progressively during cold exposure . Transactivation assays in HIB 1B and P29320 -293 cells demonstrated an adrenergic stimulation of peroxisome proliferator response element reporter activity via Q07869 , establishing a role for these nuclear receptors in hormonal regulation of gene transcription in Q14032 . The appearance of dermcidin isoform 2 , a novel platelet aggregating agent in the circulation in acute myocardial infarction that inhibits insulin synthesis and the restoration by acetyl salicylic acid of its effects . Hyperglycemia with severe reduction of plasma insulin level is frequently associated with acute ischemic heart disease . Since insulin is reported to be an anti thrombotic humoral factor , the mechanism of the impaired insulin synthesis was investigated . The plasma from the patients with acute myocardial infarction ( AMI ) was analyzed by SDS-polyacrylamide gel electrophoresis . P81605 isoform 2 ( dermcidin ) was determined by enzyme linked immunosorbent assay . P01308 synthesis was determined by in vitro translation of glucose induced insulin mRNA synthesis in the pancreatic β cells . DB00435 ( NO ) was determined by methemoglobin method . SDS-polyacrylamide gel electrophoresis of AMI plasma demonstrated the presence of a novel protein band of Mr 11 kDa that was determined to be dermcidin . Addition of 0.1 μM dermcidin inhibited insulin synthesis by > 65 fold compared to control through the inhibition of NO synthesis in the pancreatic cells . The oral administration of 150 mg acetyl salicylic acid ( aspirin ) to the AMI patients increased the plasma insulin level from 13 ( median ) to 143 μunits/dl ( median ) with concomitant decrease of plasma dermcidin level from 112 to 9 nM in these patients within 12 h . It was also found that while the injection of 3.0 ± 0.05 ( n = 10 ) nmol dermcidin with 0.25 ± 0.03 μmol ADP/g body weight caused coronary thrombus in mice , ADP itself at this concentration failed to produce thrombus . These results indicated that dermcidin was a novel platelet aggregating agent , and potentiated the ADP induced thrombosis in the animal model as well as acutely inhibited glucose induced insulin synthesis .

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DB00107 alleviates the neuroendocrine and cytokine response to bacterial endotoxin in healthy men . DB00107 is a hormone and neurotransmitter found to have anti-inflammatory functions in rodents . Here we used experimental bacterial endotoxinemia to examine the role of exogenous oxytocin administration on innate immune responses in humans . Ten healthy men received , in a randomized , placebo-controlled , crossover design , placebo , oxytocin , LPS , and LPS + oxytocin . DB00107 treatment resulted in a transient or prolonged reduction of endotoxin-induced increases in plasma DB01285 , cortisol , procalcitonin , P01375 , IL-1 receptor antagonist , P05112 , P05231 , macrophage inflammatory protein-1alpha , macrophage inflammatory protein-1beta , monocyte chemoattractant protein-1 ( P13500 ) , interferon-inducible protein 10 , and P15692 . In vitro , oxytocin had no impact on LPS effects in releasing P01375 , P05231 , and P13500 in monocytes and peripheral blood mononuclear cells from healthy human donors . In summary , oxytocin decreases the neuroendocrine and cytokine activation caused by bacterial endotoxin in men , possibly due to the pharmacological modulation of the cholinergic anti-inflammatory pathway . DB00107 might be a candidate for the therapy of inflammatory diseases and conditions associated with high cytokine and P15692 levels . Acute-phase response proteins are related to cachexia and accelerated angiogenesis in gastroesophageal cancers . BACKGROUND : Accurate outcome prediction in gastroesophageal malignancies is challenging . Acute-phase response proteins ( APRPs ) have been claimed to be independent prognosticators , although the basis for their association with prognosis remains unexplained . We hypothesized that , similarly to pancreatic and lung cancers , changes in APRPs in gastroesophageal malignancies are associated with cachexia and accelerated angiogenesis . METHODS : P02741 ( CRP ) , albumin and transferrin serum levels were evaluated and the Glasgow Prognostic Score ( GPS ) calculated . These data were compared with concentrations of circulating interleukin ( IL ) -1 , P05231 and P10145 , tumor necrosis factor-alpha ( P01375 ) , vascular endothelial growth factor ( P15692 ) -A , P49767 and midkine in 96 gastroesophageal cancer patients ( 49 with cachexia ) and 42 healthy subjects . RESULTS : Albumin and CRP levels were altered in the cancer patients , with further CRP elevation in those with cachexia . P02787 was decreased only in the cachectic patients . The interrelationships between the APRPs were strengthened in cachexia and only then were APRPs correlated with the cytokines elevated in gastroesophageal cancer-related cachexia : P05231 , P10145 , P15692 and midkine . GPS corresponded well to transferrin , IL-1 , P05231 , P10145 , P01375 , P15692 and midkine concentrations . CONCLUSIONS : Cachexia in gastroesophageal cancers is associated with changes in APRP concentrations . This , together with a direct relationship of APRPs with accelerated angiogenesis , may constitute a foundation for the association of APRPs and GPS with outcome in these malignancies . Cysteinyl leukotrienes enhance tumour necrosis factor-alpha-induced matrix metalloproteinase-9 in human monocytes/macrophages . BACKGROUND : P14780 ( P14780 ) is an important enzyme responsible for airway remodelling . Monocytes/macrophages have a cysteinyl leukotriene 1 ( cysLT1 ) receptor , but its function is poorly understood . OBJECTIVE : To elucidate the function of the cysLT1 receptor of human monocytes/macrophages in P14780 production . METHODS : We examined the effect of cysLTs ( LTC4 , -D4 and -E4 ) on P01375 -induced P14780 production in THP-1 cells , a human monocytic leukaemia cell line and peripheral blood P08571 + monocytes/macrophages . In addition , we examined the effect of pranlukast , a cysLT1 receptor antagonist , on the enhancement of P01375 -induced P14780 production by cysLTs . RESULTS : ELISA revealed that LTC4 and -D4 , but not -E4 , enhanced P01375 -induced P14780 production in THP-1 cells and peripheral blood P08571 + monocytes/macrophages . Real-time polymerase chain reaction demonstrated that LTC4 and -D4 , but not -E4 , increased P14780 mRNA expression induced by P01375 in THP-1 cells . Moreover , we demonstrated that pranlukast completely inhibited the enhancement of P01375 -induced P14780 production by LTC4 and -D4 in THP-1 cells and peripheral blood P08571 + monocytes/macrophages . CONCLUSION : LTC4 and -D4 enhanced the P01375 -induced P14780 production via binding the cysLT1 receptor in human monocytes/macrophages . DB01411 inhibited the enhancements by LTC4 and D4 . DB01370 alters iron and manganese uptake and regulation of surface transferrin receptors in primary rat oligodendrocyte cultures . P02787 ( Tf ) is a major transport protein for both iron ( Fe ) and aluminum ( Al ) , as well as manganese ( Mn ) and it can mediate cellular uptake of these elements via cell surface Tf receptors . To study the effect of Al-Tf on Tf receptor regulation , primary oligodendrocyte cultures were prepared from cortices of newborn rats . The effects of Al-Tf on 54Mn and 59Fe uptake were compared to those of apo- , Fe- , or Mn-Tf ( 1.25 microM ) . To examine changes in cell surface Tf binding capacity , preincubation ( 4 h , 37 degrees C ) was performed with apo- , Al- or Fe-Tf and homologous receptor binding studies were subsequently conducted with 125I-Fe-Tf at 4 degrees C . Incubation with Al-Tf , but not with equimolar amounts of Al chloride or Al citrate , led to dose-related increases in cellular Al . Incubation with either Al- or Fe-Tf decreased 59Fe uptake , while incubation with either Al- or Mn-Tf decreased 54Mn uptake . Surface Tf receptor sites/cell were 1.05 , 0.60 and 0.46 x 10(5) after incubations with equivalent amounts of apo- , Fe- , and Al-Tf respectively . The data suggest that Al-Tf down-regulates surface Tf receptors on oligodendrocytes and can limit Fe and Mn uptake through this mechanism . 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 . Uremic Toxins Induce ET-1 Release by Human Proximal Tubule Cells , which Regulates Organic Cation Uptake Time-Dependently . In renal failure , the systemic accumulation of uremic waste products is strongly associated with the development of a chronic inflammatory state . Here , the effect of cationic uremic toxins on the release of inflammatory cytokines and endothelin-1 ( ET-1 ) was investigated in conditionally immortalized proximal tubule epithelial cells ( ciPTEC ) . Additionally , we examined the effects of ET-1 on the cellular uptake mediated by organic cation transporters ( OCTs ) . Exposure of ciPTEC to cationic uremic toxins initiated production of the inflammatory cytokines P05231 ( 117 ± 3 % , p < 0.001 ) , P10145 ( 122 ± 3 % , p < 0.001 ) , and ET-1 ( 134 ± 5 % , p < 0.001 ) . This was accompanied by a down-regulation of O75051 mediated 4-(4-(dimethylamino)styryl)-N-methylpyridinium-iodide ( ASP+ ) uptake in ciPTEC at 30 min ( 23 ± 4 % , p < 0.001 ) , which restored within 60 min of incubation . Exposure to ET-1 for 24 h increased the ASP+ uptake significantly ( 20 ± 5 % , p < 0.001 ) . These effects could be blocked by BQ-788 , indicating activation of an P24530 -receptor-mediated signaling pathway . Downstream the receptor , P35228 inhibition by ( N(G)-monomethyl-l-arginine ) l-NMMA acetate or aminoguanidine , as well as protein kinase C activation , ameliorated the short-term effects . These results indicate that uremia results in the release of cytokines and ET-1 from human proximal tubule cells , in vitro . Furthermore , ET-1 exposure was found to regulate proximal tubular O75051 transport activity in a differential , time-dependent , fashion . Experimental Staphylococcus aureus infection of the mammary gland induces region-specific changes in innate immune gene expression . Staphylococcus aureus is a prolific mastitis-causing bacterium that resides naturally in the environment of the dairy cow . The aim of this study was to profile immune gene expression in tissue from the alveolar , ductal , gland cistern and teat canal regions of the bovine mammary gland following intramammary infection with S. aureus . Quantitative real-time PCR ( qPCR ) was used to profile expression of innate immune genes including pattern recognition receptors ( PRRs ) , cytokines , antimicrobial peptides ( AMPs ) and acute phase proteins ( APPs ) . Consistent expression of Toll-like receptors ( TLRs ) 1-10 and NOD-like receptors ( NODs ) 1-2 was detected in all four tissue regions . Pro-inflammatory cytokines ( P05231 , Q16552 and P10145 ) and anti-inflammatory cytokine ( P22301 ) were induced in all 4 tissues . P05067 ( SAA3 and HP ) and AMP ( DEFB4 and Q8NG35 ) genes showed the greatest induction throughout the mammary gland in response to S. aureus , with particularly high expression in alveolar tissue ( SAA3 and HP > 133- and > 80-fold respectively , P < 0.05 ; DEFB4 and Q8NG35 > 9- and > 27-fold respectively , P < 0.05 ) . Collectively , our data show both sentinel and effector immune functions throughout the mammary gland in response to S. aureus challenge . Favorable coagulation profile with fondaparinux after hip surgery in elderly patients . Twenty-three patients with fondaparinux prophylaxis over 75 years of age who underwent hip fracture surgery were enrolled in the study . DB00569 ( 2.5 mg ) was administered subcutaneously 6 h postoperatively and then every 24 h for 28 days . Coagulation and inflammatory parameters were measured preoperatively , then 10 h , 2 , 7 , and 28 days postoperatively . Increased D-dimers , positive acute phase proteins , and P05231 , and decreased negative acute phase proteins were observed preoperatively ( P < 0.05 ) . Maximum values were reached 10 h postoperatively for P05231 and D-dimer , and on postoperative days 2 and 7 for positive acute phase proteins ( P < 0.05 ) . P02787 , prealbumin and antithrombin levels were lowest 10 h postoperatively and on postoperative day 2 ( P < 0.05 ) . Increased D-dimers , P05231 , and positive acute phase proteins , and decreased negative acute phase proteins persisted until postoperative day 28 ( P < 0.05 ) . P00734 fragments ( F1 + 2 ) reached peak levels preoperatively and decreased gradually until postoperative day 28 . Fondaparinux promoted the inhibition of thrombin generation , as documented by negative correlation between F1 + 2 and FXa inhibition ( r = -0.46 ; P < 0.001 ) . Fondaparinux-induced FXa inhibition increased gradually until postoperative day 28 . This increase correlated positively with antithrombin activity ( r = 0.4 ; P < 0.05 ) . Fondaparinux prophylaxis counteracted pro-thrombogenic effect associated with hip fracture and subsequent surgery without severe bleeding complications . Overproduced interleukin 6 decreases blood lipid levels via upregulation of very-low-density lipoprotein receptor . BACKGROUND : Interleukin 6 ( P05231 ) blockade raises blood lipid levels in patients with rheumatoid arthritis . OBJECTIVE : To examine the influence of P05231 on lipid metabolism . METHODS : Vascular smooth muscle cells ( VSMC ) were cultured in the presence of P05231 , soluble P05231 receptor ( sIL6R ) , P05231 +sIL6R or tumour necrosis factor alpha ( TNFalpha ) for 24 h . After culture , the expression of very-low-density lipoprotein receptor ( P98155 ) , low-density lipoprotein receptor ( P01130 ) and low-density lipoprotein-related protein-1 ( Q07954 ) were measured by real-time PCR . Human P05231 was injected into mice twice a day for 2 weeks and then P98155 expression in several tissues and the change of total cholesterol ( TC ) and triglyceride ( TG ) levels were investigated . Finally , the effect of anti- P05231 receptor ( P08887 ) antibody injection on blood lipid levels was examined . RESULTS : P05231 +sIL6R significantly induced expression of P98155 mRNA in VSMC ( 8.6-fold , p < 0.05 ) , but P05231 or sIL6R alone and TNFalpha did not do so . None of these cytokines induced P01130 and Q07954 mRNA expression . P05231 injection into mice increased the expression of P98155 in heart , adipose tissue and liver and decreased TC and TG levels . The injection of anti- P08887 antibody normalised the reduced levels of TC and TG caused by P05231 injection , whereas it had no influence on the levels of TC and TG in normal mice . CONCLUSIONS : Overproduced P05231 decreased blood lipid levels by increasing P98155 expression in several tissues . It is concluded that P05231 blockade normalises reduced lipid levels caused by P05231 , but does not affect normal lipid metabolism . Effects on thrombin generation of single injections of DB02351 in patients with calf vein thrombosis . STUDY OBJECTIVE : To determine whether single injections of DB02351 , a direct thrombin inhibitor , can inhibit thrombin generation in patients with calf vein thrombosis and , if so , if the inhibition is sustained . DESIGN : Phase II open label cohort study . SETTING : Tertiary-care referral centres , university affiliated hospitals . PATIENTS : 10 patients with venographically-demonstrated calf vein thrombosis . INTERVENTION : Patients received a single injection of DB02351 , either 1.0 mg/kg subcutaneously or 0.6 mg/kg as a 15 min intravenous infusion . P00734 fragment ( F1++2 ) levels , as an index of thrombin generation , were measured before as well as 6 h post- and 24 h post- DB02351 administration . Patients were followed with non-invasive tests to detect thrombus extension into the proximal veins . RESULTS : There was a significant reduction in the levels of F1+2 with both regimens , 6 h after DB02351 . The F1+2 levels 24 h post- DB02351 showed a significant increase relative to the 6 h post- DB02351 results . One patient developed thrombus extension into the popliteal vein and was treated with conventional anticoagulants . CONCLUSION : The single injections of DB02351 used in the study produced incomplete and temporary suppression of F1+2 . Complete and permanent inhibition of thrombin generation with DB02351 in patients with calf vein thrombosis may require higher doses , multiple subcutaneous injections and/or prolonged intravenous infusion . P50579 is required for P19526 initiation and proliferation . In a chemical screening , we tested the antiangiogenic effects of fumagillin derivatives and identified fumagillin as an inhibitor of definitive hematopoiesis in zebrafish embryos . DB02640 is known to target methionine aminopeptidase II ( MetAP2 ) , an enzyme whose function in hematopoiesis is unknown . We investigated the role of MetAP2 in hematopoiesis by using zebrafish embryo and human umbilical cord blood models . Zebrafish metap2 was expressed ubiquitously during early embryogenesis and later in the somitic region , the caudal hematopoietic tissue , and pronephric duct . metap2 was inhibited by morpholino and fumagillin treatment , resulting in increased mpo expression at 18 hours postfertilization and reduced c-myb expression along the ventral wall of dorsal aorta at 36 hours postfertilization . It also disrupted intersegmental vessels in Tg(fli1:gfp) embryos without affecting development of major axial vasculatures . Inhibition of MetAP2 in CB P28906 (+) cells by fumagillin had no effect on overall clonogenic activity but significantly reduced their engraftment into immunodeficient nonobese diabetes/severe combined immunodeficiency mice . metap2 knock-down in zebrafish and inhibition by fumagillin in zebrafish and human CB P28906 (+) cells inhibited P62158 Kinase II activity and induced P29323 phosphorylation . This study demonstrated a hitherto-undescribed role of MetAP2 in definitive hematopoiesis and a possible link to noncanonical Wnt and P29323 signaling . 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 , is a selective , non-competitive inhibitor of acetylcholinesterase ( P22303 ) . In vivo , DB04892 produces rapid , potent , and long-lasting P22303 inhibition . As a possible result of its preferential brain selectivity , DB04892 is significantly less toxic than (-)-physostigmine . In studies using the Stone maze paradigm , DB04892 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 appears to have a second mode of action . Reduced secretion of beta-amyloid ( Abeta ) has been observed in cell lines exposed to DB04892 , occurring through translational regulation of beta-amyloid precursor protein ( beta- P05067 ) 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 treatment tended to reduce beta- P05067 and Abeta levels in plasma samples . Clinical studies also reveal that DB04892 ( 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 in patients with AD are currently ongoing . 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 . The anti-androgen drug dutasteride renders triple negative breast cancer cells more sensitive to chemotherapy via inhibition of HIF-1α-/ P15692 -signaling . BACKGROUND : Triple negative breast cancer ( TNBC ) is characterized by lack of expression of both estrogen and progesterone receptor as well as lack of amplification of P04626 . Patients with TNBC carry an unfavorable prognosis compared to other breast cancer subtypes given that endocrine or P04626 targeted therapies are not effective , rendering chemotherapy the sole effective treatment option to date . Therefore , there is a high demand for additional novel treatment options . FINDINGS : We previously published a list of genes showing both higher gene expression rates in TNBC and , in addition , are known to encode targets of non-oncologic drugs . P18405 , which encodes the type-1 isoform of the steroid-5alpha-reductase , which is involved in androgen metabolism , was found to be one of these genes . DB01126 is a dual blocker of both the type-1 and type-2 isoform of P18405 and is indicated in the treatment of benign prostate hyperplasia . Treatment of TNBC cell lines with dutasteride was associated with a dose-dependent decrease in cell viability , altered protein expression of P15692 and HIF-1α and increased chemosensitivity . CONCLUSION : Our results demonstrate that the P18405 -corresponding anti-androgenic drug dutasteride might act as a combinatorial therapeutic option besides standard chemotherapy in highly aggressive TNBC . Increased P05231 levels in pituitary-deficient patients are independently related to their carotid intima-media thickness . OBJECTIVE : Increased cardiovascular morbidity and mortality has been observed in patients with pituitary deficiency and untreated growth hormone deficiency ( GHD ) . We investigated peripheral inflammatory and fibrinolytic markers and their associations with arterial intima-media thickness ( IMT ) in GHD . DESIGN : Cross-sectional study . PATIENTS : Thirty-four patients with GHD , but without cardiovascular disease , were compared to healthy controls matched for age , sex , body mass index ( BMI ) and smoking habits . MEASUREMENTS : IMT of the common carotid artery , P02741 ( CRP ) , interleukin-6 ( P05231 ) , fibrinogen , plasminogen activator inhibitor-1 ( P05121 ) activity and tissue plasminogen activator antigen ( tPA-ag ) were measured . RESULTS : Median P05231 concentrations were increased by 208 % and 248 % in GHD patients compared to BMI-matched and nonobese controls , respectively . Median CRP and tPA-ag levels were increased by 237 % and 167 % in patients compared to nonobese controls , but not significantly different compared to BMI-matched controls . Plasma levels of fibrinogen and P05121 activity did not differ between groups . Age , low-density lipoprotein ( LDL ) cholesterol , tPA-ag and P05231 were positively correlated , and P05019 was negatively correlated to IMT in the patient group , but only age and P05231 were independently related to IMT . CONCLUSIONS : P05231 concentrations were increased in GHD patients compared to controls and independently related to IMT in patients . This finding may help to explain the variance in IMT and the increased vascular morbidity and mortality in hypopituitary patients with GHD . P55157 inhibitor decreases plasma cholesterol levels in P01130 -deficient WHHL rabbits by lowering the VLDL secretion . To examine whether a microsomal triglyceride transfer protein ( P55157 ) -inhibitor is effective in patients with homozygous familial hypercholesterolemia , we administered ( 2S ) -2-cyclopentyl-2-[4-[(2,4-dimethyl-9H-pyrido[2,3-b]indol-9-yl)methyl]phenyl]-N- [ ( 1S ) -2-hydroxy-1-phenylethyl ] ethanamide ( DB04852 ) , a new P55157 inhibitor , to low-density lipoprotein ( LDL ) -receptor-deficient Watanabe heritable hyperlipidemic ( WHHL ) rabbits at doses of 3 , 6 , and 12 mg/kg for 4 weeks . In the 12 mg/kg group , the plasma cholesterol and triglyceride levels were decreased by 70 % and 45 % , respectively , and the very low-density lipoprotein ( VLDL ) secretion rate was decreased by 80 % . The composition of newly secreted VLDL was similar in each group . This suggests that DB04852 diminished the number of VLDL particles secreted from the liver . Although the ratio of vitamin E/LDL was not altered by DB04852 , triglyceride accumulation and a decrease in vitamin E were observed in the liver . In conclusion , an inhibition of VLDL secretion led to a decrease of plasma LDL in WHHL rabbits , and P55157 inhibitors should have hypolipidemic effects against homozygous familial hypercholesterolemia . Vitamin D analogues . The plethora of actions attributed to 1,25(OH)2D3 throughout the body have suggested potential therapeutic applications for the treatment of hyperproliferative diseases , immune dysfunction , endocrine disorders , and metabolic bone disease . However , the potent calcemic activity of the natural vitamin D hormone has precluded its use in most cases . New vitamin D analogues are under development that display greater specificity , in most cases , by retaining the therapeutic properties of 1,25(OH)2D3 , but with lower calcemic activity . Two analogues have been approved for use in patients : calcipotriol ( DB02300 from Leo Pharmaceuticals , Copenhagen , Denmark ) for the treatment of psoriasis ; and 19-nor-1,25(OH)2D2 ( DB00910 from Abbott Laboratories , Abbott Park , IL ) for secondary hyperparathyroidism . Many others analogues are currently being tested in preclinical and clinical trials for the treatment of various types of cancer and osteoporosis , and for immunosuppression . The selectivity of the analogues can be attributed to the combined interactions with four proteins : the vitamin D receptor ( P11473 ) , the serum vitamin D binding protein ( DBP ) , the vitamin D-24-hydroxylase and to a newly described membrane receptor . Low DBP affinity has been shown to be responsible for the reduced calcemic actions of calcipotriol and 22-oxacalcitriol ( O75051 ) , which is being tested for secondary hyperparathyroidism . However , the low calcemic activity of other analogues , including 19-nor-1,25(OH)2D2 , involves other , as yet undefined , mechanisms . Understanding of the molecular basis for the selectivity of the vitamin D analogues will allow the design of more effective and safer vitamin D compounds for the treatment of a wide range of clinical disorders . P01308 action on H292 bronchial carcinoma cells as compared to normal bronchial epithelial cells . DB00030 may contribute to bronchial carcinoma due to P08069 activation by high local concentrations . Therefore , effects of insulin and P05019 on human bronchial carcinoma cells ( H292 ) and normal bronchial epithelium cells ( P02100 ) were studied . TGF-β was included since it also influences carcinoma progression . H292 and P02100 cells expressed both the insulin receptor and the P08069 ; in H292 cells an additional , shorter , splicing variant ( IR-A ) of the insulin receptor was present . P06213 expression was around four to five times higher in H292 than in P02100 cells at mRNA and protein levels . P01308 and TGF-β exerted contrary actions on proliferation and gene expression in H292 cells . Genes regulated by insulin , P05019 , and TGF-β were linked to inflammation , cell adhesion , muscle contraction and differentiation . P01308 and P05019 also suppressed DNA repair genes . EC(50) for insulin-induced proliferation was around 5 nM in H292 and around 30 nM P02100 cells . The EC(50) values for gene expression ranged from 9 to 90 nM in both cell types , dependent on the gene studied . In H292 cells , the proliferative response was much stronger if TGF-β was present . In P02100 cells this interaction of insulin and TGF-β was not observed , and changes in gene expression were mostly lower by at least 10-fold as compared to H292 . All in all , the insulin effects in H292 were generally much stronger than in P02100 cells and - with regard to proliferation - occurred at lower concentrations . Thus , insulin will hardly induce cancer from normal bronchial cells but may favour progression of pre-existing tumours .

MH_train_1328

MH_train_1328

MH_train_1328

interacts_with DB00333?

multiple_choice

Association of polymorphisms in nicotinic acetylcholine receptor alpha 4 subunit gene ( P43681 ) , mu-opioid receptor gene ( P35372 ) , and ethanol-metabolizing enzyme genes with alcoholism in Korean patients . Findings obtained from several studies indicate that ethanol enhances the activity of alpha4beta2 neuronal nicotinic acetylcholine receptor and support the possibility that a polymorphism of the nicotinic acetylcholine receptor alpha4 subunit gene ( P43681 ) modulates enhancement of nicotinic receptor function by ethanol . To identify the association between the CfoI polymorphism of the P43681 and alcoholism , we examined distribution of genotypes and allele frequencies in Korean patients diagnosed with alcoholism ( n = 127 ) and Korean control subjects without alcoholism ( n = 185 ) with polymerase chain reaction-restriction fragment length polymorphism methods . We were able to detect the association between the CfoI polymorphism of the P43681 and alcoholism in Korean patients ( genotype P = .023 ; allele frequency P = .047 ) . The genotypes and allele frequencies of known polymorphisms in other alcoholism candidate genes , such as alcohol metabolism-related genes [ alcohol dehydrogenase 2 ( P00325 ) , aldehyde dehydrogenase 2 ( P05091 ) , alcohol dehydrogenase 3 ( P00326 ) , and cytochrome P450 2E1 ( P05181 ) ] and mu-opioid receptor gene ( P35372 ) , were studied . The polymorphisms of P00325 , P05091 , and P05181 were significantly different in Korean patients with alcoholism and Korean control subjects without alcoholism , but P00326 and P35372 did not differ between the two groups . Production of an antiserum specific to the ADP-ribosylated form of elongation factor 2 from archaebacteria and eukaryotes . An antiserum to ADP-ribosylated elongation factor 2 ( DB02059 - P13639 ) from S. acidocaldarius was raised in rabbits using stained , homogenized , DB02059 - P13639 -containing slices from SDS-gels as a source of antigen . P13639 ( P13639 ) from S. acidocaldarius was cloned in E. coli and the expressed gene product was used in order to adsorb all anti- P13639 antibodies which do not contain the ADP-ribosyl group within their epitopes , as E. coli is unable to synthesize the ADP-ribosyl acceptor diphthamide . The remaining antibodies were specific to ADP-ribosylated P13639 from Thermoplasma acidophilum , S. acidocaldarius and Desulfurococcus mucosus . ADP-ribosylated P13639 from eukaryotic sources also reacted with the adsorbed antiserum as shown for P13639 isolated from the killi-fish Cynolebias whitei , the mouse species BALB/c and Han/Wistar rats . The adsorbed antiserum did not cross-react with ADP-ribosylated actin or rho protein or with DB03147 -containing D-amino acid oxidase . Polymorphisms associated with egg number at 300 days of age in chickens . We looked for variations that could be associated with chicken egg number at 300 days of age ( EN300 ) in seven genes of the hypothalamic-pituitary-gonadal axis , including gonadotrophin-releasing hormone-I ( DB00644 ) , P30968 ( GnRHR ) , neuropeptide Y ( P01303 ) , dopamine D2 receptor ( P14416 ) , vasoactive intestinal polypeptide ( P01282 ) , P01282 receptor-1 ( VIPR-1 ) , prolactin ( PRL ) , and the QTL region between 87 and 105 cM of the Z chromosome . Ten mutations in the seven genes were chosen to do marker-trait association analyses in a population comprising 1310 chickens , which were obtained from a company located in Guangdong Province of China . The C1704887T of VIPR-1 was found to have a highly significant association with EN300 . The T5841629C of P14416 and the C1715301T of VIPR-1 were significantly associated with EN300 . A highly significant association was also found between the C1704887T-C1715301T haplotypes of VIPR-1 and EN300 . H1H3 had the highest EN300 . Four PCR-RFLP variations in the candidate QTL region were selected to investigate their genetic effects on EN300 . The haplotypes of T32742468C-G32742603A in this region showed a highly significant association with EN300 . Bioinformatics analyses showed that both T32742468C and G32742603A were located in intron 1 of the SH3-domain P62993 -like 2 ( Q99962 ) gene . We conclude that five SNPs , including C1704887T and C1715301T of VIPR-1 , T5841629C of P14416 , and T32742468C and G32742603A of Q99962 , would be useful as markers for breeding to increase chicken EN300 . Multiple cholinergic nicotinic receptor genes affect nicotine dependence risk in African and European Americans . Several independent studies show that the chromosome 15q25.1 region , which contains the P30532 - P32297 - P30926 gene cluster , harbors variants strongly associated with nicotine dependence , other smoking behaviors , lung cancer and chronic obstructive pulmonary disease . We investigated whether variants in other cholinergic nicotinic receptor subunit ( CHRN ) genes affect the risk of nicotine dependence in a new sample of African Americans ( AAs ) ( N = 710 ) . We also analyzed this AA sample together with a European American ( EA ) sample ( N = 2062 , 1608 of which have been previously studied ) , allowing for differing effects in the two populations . Cases are current nicotine-dependent smokers and controls are non-dependent smokers . Variants in or near Q07001 - P07510 , P36544 and Q9GZZ6 show modest association with nicotine dependence risk in the AA sample . In addition , P43681 , Q05901 - Q15825 and P11230 show association in at least one population . P07510 and P43681 harbor single nucleotide polymorphisms ( SNPs ) that have opposite directions of effect in the two populations . In each of the population samples , these loci substantially increase the trait variation explained , although no loci meet Bonferroni-corrected significance in the AA sample alone . The trait variation explained by three key associated SNPs in P30532 - P32297 - P30926 is 1.9 % in EAs and also 1.9 % in AAs ; this increases to 4.5 % in EAs and 7.3 % in AAs when we add six variants representing associations at other CHRN genes . Multiple nicotinic receptor subunit genes outside chromosome 15q25 are likely to be important in the biological processes and development of nicotine dependence , and some of these risks may be shared across diverse populations . Production and characterization of antibodies to gonadotropin-releasing hormone receptor . Antibodies to the gonadotropin-releasing hormone ( DB00644 ) receptor of bovine pituitary membranes have been raised in rabbits by immunization with affinity-purified receptor preparations . These antibodies did not compete with 125I-labeled DB00644 analog ( DB06719 ) for binding to the receptors but did precipitate rat and bovine solubilized receptors labeled with 125I- DB06719 . Binding of the antibodies to the receptors was also demonstrated by immunoprecipitation of 125I-labeled purified receptors and photoaffinity-labeled receptors . The antibodies did not have a DB00644 -like activity but rather inhibited , in a dose-dependent manner , DB00644 -stimulated luteinizing hormone release from cultured rat pituitary cells . In addition , the antibodies did not inhibit luteinizing hormone release stimulated by high K+ concentration . This suggests that the antibodies recognize domains of the receptor other than the binding site of the hormone and thereby inhibit the biological response . These P30968 antibodies provide a useful tool for studying P30968 structure , function , localization , and biosynthesis . Inhibition of canine exocrine pancreatic secretion by peptide YY is mediated by P10082 -preferring P28062 receptors . It is still unclear , which receptor subtype , Q03519 and/or P28062 , mediates the inhibitory action of P10082 on exocrine pancreatic secretion . The present study was undertaken to characterize functionally the Y receptor subtype that mediates the inhibition of exocrine pancreatic secretion by peptide YY ( P10082 ) . In eight conscious dogs with chronic gastric and pancreatic fistulas , we compared the action of intravenous infusion of 200 and 400 pmol/kg/h of the Y receptor agonists P10082 1-36 , DB05004 , P10082 13-36 , Pro34PYY 1-36 , and P01303 1-36 on the pancreatic secretory response to secretin ( 20.5 pmol/kg/h ) and cerulein ( 29.6 pmol/kg/h ) . P10082 13-36 , Pro34PYY 1-36 , and P01303 1-36 were also studied by giving a fivefold dose ( 1,000 and 2,000 pmol/kg/h ) . P10082 1-36 and the P49146 agonist DB05004 significantly inhibited pancreatic secretory responses to secretin and cerulein , whereas inhibition by P01303 1-36 and the P49146 agonist P10082 13-36 was attainable only at doses of 1,000 and 2,000 pmol/kg/h . The Q03519 receptor agonist Pro34PYY 1-36 was without effect on pancreatic secretion . We conclude that in dogs the inhibition of exocrine pancreatic secretion by P10082 is mediated via P28062 receptors of a P10082 -preferring subtype . Lack of evidence for the mu-opioid receptor splice variant MOR1C in rats . We previously reported the existence of P35372 (C) mRNA and P35372 (C)-immunoreactivity ( -ir ) in rats . However , the sequence that we reported for rat P35372 (C) appears not to be present in the rat genome . We have therefore reexamined whether P35372 (C) mRNA or P35372 (C)-ir exist in rats . We used reverse-transcription polymerase chain reaction ( RT-PCR ) to attempt to amplify P35372 , P35372 (A) , P35372 (B) , the rat P35372 (C) sequence we previously reported , and P35372 (C1) and P35372 ( P06681 ) ( which have recently been reported to exist in rats ) . In RNA extracted from rats , we were able to demonstrate PCR products representing P35372 , P35372 (A) , and P35372 (B) splice variants . All three products were confirmed as related to P35372 by Southern blot . However , we were unable to detect either the P35372 (C) product reported previously by us or the P35372 (C)-like products reported to exist in rats by others . In RNA extracted from mice we were able to detect P35372 , P35372 (A) , P35372 (B) , and P35372 (D)-like products . To test the specificity of our P35372 (C) antiserum , we examined P35372 (C)-ir in control and knockout mice . P35372 (C)-ir had a distribution in control mice similar to that previously reported in rats , including coexisting with vGLUT2 . However , although P35372 -ir was absent in P35372 knockout mice , the density and distribution of P35372 (C)-ir were unchanged , suggesting that the antiserum crossreacts with another molecule in tissue . We find no evidence for P35372 (C) mRNA in rats . Furthermore , we conclude that P35372 (C)-ir represents crossreactivity . Role of antispasmodics in the treatment of irritable bowel syndrome . Irritable bowel syndrome ( IBS ) is a long-lasting , relapsing disorder characterized by abdominal pain/discomfort and altered bowel habits . Intestinal motility impairment and visceral hypersensitivity are the key factors among its multifactorial pathogenesis , both of which require effective treatment . Voltage-gated calcium channels mediate smooth muscle contraction and endocrine secretion and play important roles in neuronal transmission . Antispasmodics are a group of drugs that have been used in the treatment of IBS for decades . DB01616 citrate , a spasmolytic , decreases the sensitivity of smooth muscle contractile proteins to calcium , and it is a selective P08908 receptor antagonist . DB01616 , in combination with simethicone , has been demonstrated to effectively reduce abdominal pain and discomfort in a large placebo-controlled trial . Mebeverine is a musculotropic agent that potently blocks intestinal peristalsis . Non-placebo-controlled trials have shown positive effects of mebeverine in IBS regarding symptom control ; nevertheless , in recent placebo-controlled studies , mebeverine did not exhibit superiority over placebo . Otilonium bromide is poorly absorbed from the GI tract , where it acts locally as an L-type calcium channel blocker , an antimuscarinic and a tachykinin NK2 receptor antagonist . Otilonium has effectively reduced pain and improved defecation alterations in placebo-controlled trials in IBS patients . DB09090 bromide is also an L-type calcium channel blocker that acts locally in the GI tract . DB09090 improves motility disorders and consequently reduces stool problems in IBS patients . Phloroglucinol and trimethylphloroglucinol are non-specific antispasmodics that reduced pain in IBS patients in a placebo-controlled trial . Antispasmodics have excellent safety profiles . T-type calcium channel blockers can abolish visceral hypersensitivity in animal models , which makes them potential candidates for the development of novel therapeutic agents in the treatment of IBS . Assessment of partially deoxygenated deoxynojirimycin derivatives as glucosylceramide synthase inhibitors . Q16739 ( Q16739 ) 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 ) 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 -hydroxyl of the deoxynojirimycin core is important for Q16739 inhibition . Here we show that P13671 -OH appears of less important , which may set guidelines for the development of Q16739 inhibitors that have less affinity ( in comparison with DB00419 ) for other glycoprocessing enzymes , in particular those hydrolases that act on glucosylceramide . Pharmacogenomics of methadone maintenance treatment . DB00333 is the major opioid substitution therapy for opioid dependence . Dosage is highly variable and is often controlled by the patient and prescriber according to local and national policy and guidelines . Nevertheless many genetic factors have been investigated including those affecting its metabolism ( P20813 -consistent results ) , efflux transport ( P-gp-inconsistent results ) , target μ-opioid receptor ( μ-opioid receptor-inconsistent results ) and a host of other receptors ( P14416 ) and signaling elements ( P48051 and P32121 ; not replicated ) . None by themselves have been able to substantially explain dosage variation ( the major but not sole end point ) . When multiple genes have been combined such as P08183 , P20813 , P35372 and P14416 a greater contribution to dosage variation was found but not as yet replicated . As stabilization of dosage needs to be made rapidly , it is imperative that larger internationally based studies be instigated so that genetic contribution to dosage can be properly assessed , which may or may not tailor to different ethnic groups and each country 's policy towards an outcome that benefits all . Identification of G2/M targets for the Q96HU1 kinase pathway by functional proteomics . Although the importance of the extracellular signal-regulated kinase ( P29323 ) pathway in regulating the transition from P55008 to S has been extensively studied , its role during the G2/M transition is less well understood . Previous reports have shown that inhibition of the P29323 pathway in mammalian cells delays entry as well as progression through mitosis , suggesting the existence of molecular targets of this pathway in M phase . In this report we employed 2-DE and MS to survey proteins and PTMs in the presence versus absence of Q02750 /2 inhibitor . Targets of the P29323 pathway in G2/M were identified as elongation factor 2 ( P13639 ) and nuclear matrix protein , 55 kDa ( Nmt55 ) . Phosphorylation of each protein increased under conditions of P29323 pathway inhibition , suggesting indirect control of these targets ; regulation of P13639 was ascribed to phosphorylation and inactivation of upstream P13639 kinase , whereas regulation of Nmt55 was ascribed to a delay in normal mitotic phosphorylation and dephosphorylation . 2-DE Western blots probed using anti-phospho- DB00156 -Pro antibody demonstrated that the effect of P29323 inhibition is not to delay the onset of phosphorylation controlled by cdc2 and other mitotic kinases , but rather to regulate a small subset of targets in M phase in a nonoverlapping manner with cdc2 . P35372 mutant , T394A , abolishes opioid-mediated adenylyl cyclase superactivation . This study was to characterize the effects of a point-mutant at C-terminal of mu opioid receptor ( MOR ) , namely MOR T394A , in chronic opioid-induced cellular responses . After 18 h of exposure to [ D-Ala , N-Me- DB00120 , DB00145 -ol ] enkephalin ( DAMGO ) , adenylyl cyclase ( AC ) superactivation , a hallmark for the cellular adaptive response after chronic opioid stimulation , was observed in the cells expressing wild-type receptor , but was totally abolished in the cells expressing MOR T394A . Receptor phosphorylation was also attenuated in cells with MOR T394A after prolonged preexposure to agonist . Furthermore , Q96HU1 kinase kinase-1 ( Q02750 ) overexpression was able to rescue AC superactivation in cells with MOR T394A , but showed no effect in the wild-type MOR-expressing cells . These results indicated that the amino acid T394 at C-terminus of MOR played a critical role in chronic agonist-induced AC superactivation and receptor phosphorylation . Localization of the succinate receptor in the distal nephron and its signaling in polarized MDCK cells . When the succinate receptor ( Q9BXA5 ) is activated in the afferent arterioles of the glomerulus it increases renin release and induces hypertension . To study its location in other nephron segments and its role in kidney function , we performed immunohistochemical analysis and found that Q9BXA5 is located in the luminal membrane of macula densa cells of the juxtaglomerular apparatus in close proximity to renin-producing granular cells , the cortical thick ascending limb , and cortical and inner medullary collecting duct cells . In order to study its signaling , Q9BXA5 was stably expressed in Madin-Darby Canine Kidney ( MDCK ) cells , where it localized to the apical membrane . Activation of the cells by succinate caused Gq and Gi-mediated intracellular calcium mobilization , transient phosphorylation of extracellular regulated kinase ( P29323 )1/2 and the release of arachidonic acid along with prostaglandins E2 and I2 . Signaling was desensitized without receptor internalization but rapidly resensitized upon succinate removal . Immunohistochemical evidence of phosphorylated P27361 /2 was found in cortical collecting duct cells of wild type but not Q9BXA5 knockout streptozotocin-induced diabetic mice , indicating in vivo relevance . Since urinary succinate concentrations in health and disease are in the activation range of the Q9BXA5 , this receptor can sense succinate in the luminal fluid . Our study suggests that changes in the luminal succinate concentration may regulate several aspects of renal function . Linkage assignment of eleven genes to the porcine genome . We report comparative linkage mapping of eleven genes in the swine genome by RFLP analysis . These genes include : Acid phosphatase type 5 ( P13686 ) , Cholecystokinin Type B Receptor ( P32239 ) , Antibiotic Peptide ( P49913 ) , P01308 -like Growth Factor 1 Receptor ( P08069 ) , Integrin Alpha M ( P11215 ) , Integrin Beta 2 ( ITGbeta2 ) , Opioid Receptor Mu-1 ( P35372 ) , Pro-hormone Converter ( PC1/3 ) , DB00162 Binding Q12988 ( P10745 ) , Ribosomal DNA ( RNR1 ) , and Zona Pellucida Glycoprotein 1 ( P60852 ) . The P32239 and ITGbeta2 loci define the ends of the linkage groups on Chromosomes ( Chro ) ( SSC ) 9p and 13qter , respectively . Stimulation of central β2-adrenoceptors suppresses NFκB activity in rat brain : a role for IκB . In this study we examined the impact of systemic treatment with the long-acting brain penetrant β2-adrenoceptor agonist clenbuterol on NFκB activity and IκB expression in rat brain . DB01407 decreased NFκB activity ( p65 DNA binding ) in nuclear extracts prepared from rat cortex and hippocampus for up to 8h following a single treatment . This was accompanied by increased expression of IκBα mRNA and protein . The temporal increase in IκB protein expression paralleled the suppression of NFκB activity , suggesting that IκBα mediates the suppression NFκB activity observed . These actions of clenbuterol were prevented by pre-treatment with the non-selective β-adrenoceptor antagonist propranolol , the β2-adrenoceptor antagonist ICI-118,551 , but not the β1-adrenoceptor antagonist metoprolol , suggesting that the effects of clenbuterol on IκBα expression and NFκB activity are mediated specifically by the β2-adrenoceptor . In addition , the actions of clenbuterol were mimicked by systemic administration of another highly selective long-acting β2-adrenoceptor agonist formoterol . As neurodegenerative diseases are associated with inflammation we determined if clenbuterol could suppress NFκB activation that occurs in response to an inflammatory stimulus . In this regard we demonstrate that clenbuterol inhibited IκB phosphorylation and IκB degradation and inhibited NFκB activity in hippocampus and cortex of rats following a central injection of the inflammagen bacterial lipopolysaccharide ( LPS ) . In tandem , clenbuterol blocked expression of the NFκB-inducible genes P01375 -α and P05362 following LPS administration . Our finding that clenbuterol and formoterol inhibit NFκB activity in the CNS further supports the idea that β2-adrenoceptors may be an attractive target for treating neuroinflammation and combating inflammation-related neurodegeneration . Predictive model for risk of severe gastrointestinal toxicity following chemotherapy using patient immune genetics and type of cancer : a pilot study . PURPOSE : Severe chemotherapy-induced gastrointestinal toxicity ( CIGT ) is common and results in treatment delays , dose reductions , and potential premature treatment discontinuation . Currently , there is no diagnostic marker to predict CIGT . Proinflammatory cytokines , produced via toll-like receptor signaling , are key mediators of this toxicity . Hence , this pilot study investigated the association between immune genetic variability and severe CIGT risk . METHODS : Genomic DNA from 34 patients ( 10 with severe CIGT ) who had received 5-fluoruracil-based chemotherapy regimens was analyzed for variants of IL-1B , P60568 , P05231 , IL-6R , P22301 , P01375 , TGF-b , O60603 , O00206 , Q9Y6Y9 , Q99836 , P23560 , CRP , ICE , and P35372 . Multivariate logistic regression created a prediction model of severe CIGT risk . RESULTS : There were no significant differences between patients with and without severe CIGT with regards to age , sex , type of cancer , or chemotherapy treatment regimens . The prediction model of severe CIGT risk included O60603 and P01375 genetic variability and cancer type ( colorectal and gastric ) . This prediction model was both specific and sensitive , with a receiver operator characteristic area under the curve of 87.3 % . CONCLUSIONS : This is the first report of immune genetic variability , together with cancer type , being predictive of severe CIGT risk . These outcomes are being validated in a larger patient population . Genetic , psychosocial , and demographic factors associated with social disinhibition in Mexican-origin youth . INTRODUCTION : The genetic heritability for sensation-seeking tendencies ranges from 40 to 60 % . Sensation-seeking behaviors typically manifest during adolescence and are associated with alcohol and cigarette experimentation in adolescents . Social disinhibition is an aspect of sensation-seeking that is closely tied to cigarette and alcohol experimentation . METHODS : We examined the contribution of candidate genes to social disinhibition among 1132 Mexican origin youth in Houston , Texas , adjusting for established demographic and psychosocial risk factors . Saliva samples were obtained at baseline in 2005-06 , and social disinhibition and other psychosocial data were obtained in 2008-09 . Participants were genotyped for 672 functional and tagging SNPs potentially related to sensation-seeking , risk-taking , smoking , and alcohol use . RESULTS : Six SNPs were significantly associated with social disinhibition scores , after controlling for false discovery and adjusting for population stratification and relevant demographic/psychosocial characteristics . Minor alleles for three of the SNPs ( rs1998220 on P35372 ; rs9534511 on P28223 ; and rs4938056 on O95264 ) were associated with increased risk of social disinhibition , while minor alleles for the other three SNPs ( rs1003921 on P48547 ; rs16116 downstream of P01303 ; and rs16870286 on Q8N0U6 ) exhibited a protective effect . Age , linguistic acculturation , thrill and adventure-seeking , and drug and alcohol-seeking were all significantly positively associated with increased risk of social disinhibition in a multivariable model ( P < 0.001 ) . CONCLUSIONS : These results add to our knowledge of genetic risk factors for social disinhibition . Additional research is needed to verify whether these SNPs are associated with social disinhibition among youth of different ethnicities and nationalities , and to elucidate whether and how these SNPs functionally contribute to social disinhibition . Autoantibodies against four kinds of neurotransmitter receptors in psychiatric disorders . There is a hypothesis that autoimmune abnormalities in neurotransmitter receptors might cause some psychiatric disorders . Using a sensitive radioligand assay , we detected serum autoantibodies to recombinant human muscarinic cholinergic receptor 1 ( P11229 , 34.4 % ) , mu-opioid receptor ( P35372 , 13.1 % ) , P08908 ( P08908 , 7.4 % ) , and dopamine receptor D2 ( P14416 , 4.9 % ) in 122 psychiatric patients . Positive antibodies to P11229 were found in 34.1 % , 34.9 % , 33.3 % , and 9.1 % of patients with schizophrenic disorders ( n=44 ) , mood disorders ( n=63 ) , other psychiatric disorders ( n=15 ) and autoimmune diseases ( n=33 ) , respectively . All three patients with neuroleptic maliganant syndrome had high activities of autoantibodies to P11229 , P35372 , and/or P08908 . Our data suggest that autoimmunity to neurotransmitter receptors might be associated with the induction of psychiatric symptoms and have some relation to neuroleptic malignant syndrome . The bovine 5' AMPK gene family : mapping and single nucleotide polymorphism detection . The DB00131 -activated protein kinase ( AMPK ) family is an ancient stress response system whose primary function is regulation of cellular DB00171 . Activation of AMPK , which is instigated by environmental and nutritional stresses , initiates energy-conserving measures that protect the cell by inhibition and phosphorylation of key enzymes in energy-consuming biochemical pathways . The seven genes that comprise the bovine AMPK family were mapped in cattle by using a radiation hybrid panel . The seven genes mapped to six different cattle chromosomes , each with a LOD score greater than 10.0 . Q13131 mapped to BTA 20 , P54646 and O43741 to BTA 3 , Q9Y478 to BTA 17 , P54619 to BTA 5 , Q9UGJ0 to BTA 4 , and Q9UGI9 to BTA 2 . Five of the seven genes mapped to regions expected from human/cattle comparative maps . O43741 and Q9UGI9 , however , have not been mapped in humans . We predict these genes to be located on HSA 1 and 2 , respectively . Additionally , one synonymous and one non-synonymous single nucleotide polymorphism ( SNP ) were detected in Q9UGI9 in Bos taurus cattle . In an effort to determine ancestral origins , various herds of mixed breed cattle as well as other ruminant species were characterized for sequence variation in this region of Q9UGI9 . Owing to the physiological importance of this gene family , we believe that its individual genes are candidate genes for conferring resistance to diseases in cattle . Genome-wide association analyses identify 18 new loci associated with serum urate concentrations . Elevated serum urate concentrations can cause gout , a prevalent and painful inflammatory arthritis . By combining data from > 140,000 individuals of European ancestry within the Global DB08844 Genetics Consortium ( GUGC ) , we identified and replicated 28 genome-wide significant loci in association with serum urate concentrations ( 18 new regions in or near Q7Z4K8 , P09529 , Q9UHJ3 , Q8WVE6 , P15692 , Q9UIG0 , Q9UGJ0 , P52823 , Q14541 , Q9NQ94 , Q99700 , Q8WVN8 , P08069 , O94916 , O75444 , HLF , P36896 - P37023 and Q9C0J1 ) . Associations for many of the loci were of similar magnitude in individuals of non-European ancestry . We further characterized these loci for associations with gout , transcript expression and the fractional excretion of urate . Network analyses implicate the inhibins-activins signaling pathways and glucose metabolism in systemic urate control . New candidate genes for serum urate concentration highlight the importance of metabolic control of urate production and excretion , which may have implications for the treatment and prevention of gout . Nicotinic acetylcholine receptors containing the α6 subunit contribute to ethanol activation of ventral tegmental area dopaminergic neurons . DB00184 and alcohol are often co-abused suggesting a common mechanism of action may underlie their reinforcing properties . Both drugs acutely increase activity of ventral tegmental area ( VTA ) dopaminergic ( DAergic ) neurons , a phenomenon associated with reward behavior . Recent evidence indicates that nicotinic acetylcholine receptors ( nAChRs ) , ligand-gated cation channels activated by ACh and nicotine , may contribute to ethanol-mediated activation of VTA DAergic neurons although the nAChR subtype(s) involved has not been fully elucidated . Here we show that expression and activation of nAChRs containing the α6 subunit contribute to ethanol-induced activation of VTA DAergic neurons . In wild-type ( WT ) mouse midbrain sections that contain the VTA , ethanol ( 50 or 100 mM ) significantly increased firing frequency of DAergic neurons . In contrast , ethanol did not significantly increase activity of VTA DAergic neurons in mice that do not express Q15825 , the gene encoding the α6 nAChR subunit ( α6 knock-out ( KO ) mice ) . DB00898 -induced activity in WT slices was also reduced by pre-application of the α6 subtype-selective nAChR antagonist , α-conotoxin MII[E11A] . When co-applied , ethanol potentiated the response to ACh in WT DAergic neurons ; whereas co-application of ACh and ethanol failed to significantly increase activity of DAergic neurons in α6 KO slices . Finally , pre-application of α-conotoxin MII[E11A] in WT slices reduced ethanol potentiation of ACh responses . Together our data indicate that α6-subunit containing nAChRs may contribute to ethanol activation of VTA DAergic neurons . These receptors are predominantly expressed in DAergic neurons and known to be critical for nicotine reinforcement , providing a potential common therapeutic molecular target to reduce nicotine and alcohol co-abuse . P35372 and P20813 gene variants as risk factors in methadone-related deaths . DB00333 is a medication valued for its effectiveness in the treatment of heroin addiction ; however , many fatal poisonings associated with its use have been reported over the years . We have examined the association between P20813 and micro-opioid receptor ( P35372 ) gene variations and apparent susceptibility to methadone poisoning . Genomic DNA was extracted from postmortem whole blood of 40 individuals whose deaths were attributed to methadone poisoning . The presence of P20813 *4,*9 , and *6 alleles and the P35372 A118G variant was determined by SNP genotyping . P20813 *4 , *9 , and *6 alleles were found to be associated with higher postmortem methadone concentrations in blood ( P < or = 0.05 ) . P35372 A118G was also associated with higher postmortem methadone concentrations in blood but not to a level of statistical significance ( P = 0.39 ) . In these methadone-related deaths , P35372 118GA was associated with higher postmortem benzodiazepine concentrations ( P = 0.04 ) , a finding not associated with morphine-related deaths . The risk of a methadone-related fatality during treatment may be evaluated in part by screening for P20813 *6 and A118G . DB00139 receptor Q9BXA5 provides a direct link between high glucose levels and renin release in murine and rabbit kidney . Diabetes mellitus is the most common and rapidly growing cause of end-stage renal disease in developed countries . A classic hallmark of early diabetes mellitus includes activation of the renin-angiotensin system ( DB01367 ) , which may lead to hypertension and renal tissue injury , but the mechanism of DB01367 activation is elusive . Here we identified a paracrine signaling pathway in the kidney in which high levels of glucose directly triggered the release of the prohypertensive hormone renin . The signaling cascade involved the local accumulation of succinate and activation of the kidney-specific G protein-coupled metabolic receptor , Q9BXA5 , in the glomerular endothelium as observed in rat , mouse , and rabbit kidney sections . Elements of signal transduction included endothelial Ca2+ , the production of NO and prostaglandin ( DB00917 ) , and their paracrine actions on adjacent renin-producing cells . This Q9BXA5 signaling cascade may serve to modulate kidney function and help remove metabolic waste products through renal hyperfiltration , and it could also link metabolic diseases , such as diabetes , or metabolic syndrome with DB01367 overactivation , systemic hypertension , and organ injury . mu-Opioid receptor agonists differentially regulate the expression of miR-190 and Q13562 . The agonists of mu-opioid receptor ( P35372 ) induce extracellular signal-regulated kinase ( P29323 ) phosphorylation through different pathways : morphine uses the protein kinase C ( PKC ) -pathway , whereas fentanyl functions in a beta-arrestin2-dependent manner . In addition , the two pathways result in the different cellular location of phosphorylated P29323 and the activation of different sets of transcriptional factors . In the current study , the influence of the two pathways on the expression of microRNAs ( miRNAs ) was investigated . After treating the primary culture of rat hippocampal neurons and the mouse hippocampi with morphine or fentanyl for 3 days , seven miRNAs regulated by one or two of the agonists were identified . One of the identified miRNAs , miR-190 , was down-regulated by fentanyl but not by morphine . This down-regulation was attenuated by 1,4-diamino-2,3-dicyano-1,4-bis(methylthio)butadiene ( U0126 ) , which blocks the phosphorylation of P29323 . When fentanyl-induced but not morphine-induced P29323 phosphorylation was blocked in the primary cultures from beta-arrestin2(-/-) mouse , fentanyl did not decrease the expression of miR-190 . However , a PKC inhibitor that blocked morphine-induced P29323 phosphorylation specifically had no effect on the miR-190 down-regulation . Therefore the decrease in miR-190 expression resulted from the agonist-selective P29323 phosphorylation . In addition , the expressional changes in one of the miR-190 targets , neurogenic differentiation 1 ( Q13562 ) , correlated with those in miR-190 expression , suggesting the P35372 could regulate the Q13562 pathways via the control of miR-190 expression .

MH_train_1329

MH_train_1329

MH_train_1329

interacts_with DB00502?

multiple_choice

Prenatal diagnosis of carnitine palmitoyltransferase 2 deficiency in chorionic villi : a novel approach . DB00583 palmitoyltransferase 2 ( P23786 ) deficiency , the most common autosomal recessive inherited disease of the mitochondrial long-chain fatty acid ( LCFA ) beta-oxidation , may result in three distinct clinical phenotypes , namely , a mild adult muscular form , a severe infantile hepatocardiomuscular disease , and a neonatal form , which includes dysmorphic features in addition to hepatocardiomuscular symptoms . Both the latter forms are life-threatening diseases , and prenatal diagnosis ( P01160 ) can be offered to couples at a one-fourth risk of having an affected child . P01160 of P23786 deficiency hitherto relied mostly on mutation detection from fresh chorionic villi ( 10 weeks ' gestation ) , since P23786 activity could be assayed on cultured amniocytes only ( 16-17 weeks ' gestation ) .We devised a P23786 activity assay from 10 mg of chorionic villi sampling ( CVS ) . Combining this enzymatic assay to haplotype study using polymorphic markers linked to the P23786 gene , we were able to carry out within 2 days , P23786 deficiency P01160 , in two unrelated families , using a CVS performed at the 11th week of gestation . Association study of 45 candidate genes in nicotine dependence in Han Chinese . Numerous genetic linkages , association studies have been performed in different ethnic groups and revealed many susceptibility loci and genes for nicotine dependence . However , limited similar researches were performed in Han Chinese . This study was designed to investigate the association of candidate genes with nicotine dependence in Han Chinese . We genotyped 384 SNPs within 45 candidate genes with nicotine dependence in a Han Chinese population consisting 223 high nicotine dependent subjects and 257 low nicotine dependent subjects by employing GoldenGate genotyping assay ( Illumina ) . Following association analysis was performed using PLINK software . Individual SNP-based association analysis revealed that nine SNPs located in P35462 ( rs2630351 ) , P21918 ( rs1967550 ) , Q9Y6R4 ( rs2314378 ) , DDC ( rs11575461 ) , Q05901 ( rs4954 ) , O75899 ( rs2779562 ) , P14416 ( rs11214613 and rs6589377 ) and P43681 ( rs2236196 ) were significantly associated with FTND after correction for multiple testing with the p values from 2.59×10(-7) to 9.99×10(-5) . Haplotype-based association analysis revealed haplotype G-A-A formed by rs2630351 , rs167771 and rs324032 and haplotype G-G-G-A formed by rs3773678 , rs2630349 , rs2630351 and rs167771 in P35462 ; haplotype of G-A formed by rs2779562 and rs2808566 in O75899 and haplotype of T-T-A-G-A formed by rs6832644 , rs4057797 , rs9764 , rs4552421 and rs10033119 in P25929 are associated with FTND ( p=3.61×10(-7)-8.78×10(-6) ) . Our results provided confirmation of the previous findings that P14416 , P35462 , DDC , Q05901 , O75899 and P43681 are associated with nicotine dependence . Furthermore , we for the first time report a significant association between nicotine dependence and P21918 , Q9Y6R4 and P25929 . These findings need independent replication in the future studies . DB00502 induces neurotoxicity by the DB01221 receptor downstream signaling pathway , alternative from glutamate excitotoxicity . The DB01221 receptor is believed to be important in a wide range of nervous system functions including neuronal migration , synapse formation , learning and memory . In addition , it is involved in excitotoxic neuronal cell death that occurs in a variety of acute and chronic neurological disorders . Besides of agonist/coagonist sites , other modulator sites , including butyrophenone site may regulate the N-methyl-D-aspartate receptor . It has been shown that haloperidol , an antipsychotic neuroleptic drug , interacts with the Q13224 subunit of DB01221 receptor and inhibits DB01221 response in neuronal cells . We found that DB01221 receptor was co-immunoprecipitated by anti-Ras antibody and this complex , beside NR2 subunit of DB01221 receptor contained haloperidol-binding proteins , P29475 and Ras- P01286 . Furthermore , we have shown that haloperidol induces neurotoxicity of neuronal cells via DB01221 receptor complex , accompanied by dissociation of Ras- P01286 from membranes and activation of c-Jun-kinase . Inclusion of insulin prevented relocalization of Ras- P01286 and subsequent neuronal death . DB00502 -induced dissociation of Ras- P01286 leads to inhibition of membrane-bound form of Ras protein and changes downstream regulators activity that results in the initiation of the apoptotic processes via the mitochondrial way . Our results suggest that haloperidol induces neuronal cell death by the interaction with DB01221 receptor , but through the alternative from glutamate excitotoxicity signaling pathway . Q13224 -containing DB01221 receptors as possible targets for the neuroprotective and antidepressant effects of fluoxetine . Accumulating evidence has indicated the involvement of glutamatergic neurotransmission in the pathophysiology of excitotoxicity and in the mechanism of action of antidepressants . We have previously shown that tricyclic desipramine and the selective serotonin reuptake inhibitor fluoxetine inhibit DB01221 receptors ( NMDARs ) in the clinically relevant , low micromolar concentration range . As the different subtypes of NMDARs are markedly different in their physiological and pathological functions , our aim was to investigate whether the effect of antidepressants is subtype-specific . Using whole-cell patch-clamp recordings in rat cortical cell cultures , we studied the age-dependence of inhibition of DB01221 -induced currents after treatment with desipramine and fluoxetine , as the expression profile of the NMDAR subtypes changes as a function of days in vitro . We also investigated the inhibitory effect of these antidepressants on DB01221 -induced currents in P29320 293 cell lines that stably expressed rat recombinant NMDARs with GluN1a/ Q12879 or GluN1a/ Q13224 subunit compositions . The inhibitory effect of desipramine was not age-dependent , whereas fluoxetine displayed a continuously decreasing inhibitory profile , which was similar to the Q05586 / Q13224 subtype-selective antagonist ifenprodil . In P29320 293 cells , desipramine equally inhibited DB01221 currents in both cell lines , whereas fluoxetine showed an inhibitory effect only in cells that expressed the Q05586 / Q13224 subtype . Our data show that fluoxetine is a selective inhibitor of Q13224 -containing NMDARs , whereas desipramine inhibits both Q05586 / Q12879 and Q05586 / Q13224 subtypes . As the clinical efficacy of these drugs is very similar , the putative NMDAR-associated therapeutic effect of antidepressants may be mediated only via inhibition of the Q13224 -containing subtype . The manifestation of the Q05586 / Q13224 -selectivity of fluoxetine suggests the neuroprotective potential for this drug in both acute and chronic neurodegenerative disorders . A novel inhibitory mechanism of nitrogen-containing bisphosphonate on the activity of Cl- extrusion in osteoclasts . DB09152 -containing bisphosphonates have been well known to be inhibited farnesyl diphosphate synthase ( P14324 ) , an enzyme in mevalonic acid metabolism , resulting in disturbance in polymerization of cytoskeleton structure in bone resorption and promotion of apoptosis in mature osteoclasts . Although bisphosphonates have been reported to activate ion transporters in native epithelium and Xenopus oocytes , little is known whether bisphosphonates affect acid hydrochronic acid extrusion in osteoclasts during bone resorption . The aim of this study was to determine the role of bisphosphonates on inhibition of hydrochronic acid extrusion in osteoclasts . Effects of zoledronic acid , a nitrogen-containing bisphosphonate , on the Cl(-) current activated by extracellular acidification were examined in two types of osteoclasts derived from RAW264.7 cells and mouse bone marrow macrophages ( BMMs ) . Extracellular acidification induced outwardly rectifying Cl(-) currents in mouse osteoclasts . DB00399 dose-dependently inhibited the acid-activated Cl(-) current . The non-nitrogen bisphosphonate etidronic acid had no effect on the acid-activated Cl(-) current . DB00759 -induced P14324 silencing caused a significant decrease in the Cl(-) current . The inhibitor of geranylgeranyl transferase suppressed the Cl(-) current . By contrast , the inhibitory action of zoledronic acid was rescued by addition of geranylgeranyl acid , a derivative of mevalonic acid . The activity of acid-activated Cl(-) currents was dependent on expression of P51798 during osteoclastogenesis . These results suggest that nitrogen-containing bisphosphonates suppress the activity of osteoclastic acid-activated Cl(-) currents through P14324 inhibition , suggesting the inhibition of Cl(-) extrusion activity . Clinical and genetic factors associated with nausea and vomiting in cancer patients receiving opioids . BACKGROUND : This study investigates whether demographical , disease-related and genetic factors contribute to inter-individual differences in nausea and vomiting among patients receiving opioids for cancer pain . METHODS : Cancer patients receiving opioids were included from 17 centres in 11 European countries . Intensities of nausea and vomiting were reported by 1579 patients on four-point categorical scales . In stratified regression models including demographical and disease-related factors as covariates , 96 single nucleotide polymorphisms ( SNPs ) in 16 candidate genes related to opioid- or nausea/vomiting signalling pathways ( P08183 , P35372 , P41145 , P32121 , P42226 , P21964 , P20309 , P08912 , P35367 , P14416 , P35462 , P25103 , P46098 , O95264 , Q8WXA8 , P21554 ) were analysed for association with nausea and vomiting . FINDINGS : Age , body mass index , Karnofsky Performance Status , gender , use of antiemetics , type of opioid , type of cancer and eight SNPs were associated with the inter-individual differences in nausea and vomiting among cancer patients treated with opioids ( p < 0.01 ) . The SNPs were rs1176744 , rs3782025 and rs1672717 in O95264 ; rs165722 , rs4680 and rs4633 in P21964 ; rs10802789 and rs685550 in P20309 . Only the SNP rs1672717 in O95264 passed the Benjamini-Hochberg criterion for a 10 % false discovery rate . INTERPRETATION : Clinical characteristics and SNPs within the O95264 , P21964 and P20309 genes may be associated with the variability in nausea and vomiting among cancer patients receiving opioids . This knowledge may help to identify patients at particular risk for nausea and vomiting during treatment with opioids for cancer pain . Expression of P35354 and DB01221 receptor genes at the cochlea and midbrain in salicylate-induced tinnitus . OBJECTIVE/HYPOTHESIS : The expression of the genes for cyclooxygenase ( P36551 ) and DB01221 receptor ( NR ) has seldom been reported in tinnitus . We hypothesized that expression of P35354 and NR was altered in the cochlea and midbrain in salicylate-induced tinnitus . STUDY DESIGN : Experimental study on mice . METHODS : We evaluated the tinnitus score and mRNA expression levels of P35354 and NR subtype 2B ( Q13224 ) in the cochlea and midbrain in response to intraperitoneal injections of salicylate for 4 days . RESULTS : At day 4 of tinnitus induction , the mean weights of the whole body and midbrain did not change greatly in both control and salicylate groups . The tinnitus score was not elevated from day 1 to day 4 in the control group , but increased day by day in the salicylate group . The mRNA expression level of P35354 decreased slightly in the salicylate group in the cochlea ( 1.1 ± 0.33 vs. 1.3 ± 0.49 , P = .0752 ) and in the midbrain ( 0.9 ± 0.10 versus 1.0 ± 0.35 , P = .0489 ) . Inversely , the expression levels of the Q13224 gene increased moderately in the salicylate group in the cochlea ( 3.7 ± 0.47 versus 2.3 ± 1.13 , P < 0.0001 ) and in the midbrain ( 1.6 ± 0.64 versus 1.0 ± 0.44 , P = .0007 ) . CONCLUSIONS : Salicylate induced tinnitus and altered the expression of the P35354 and Q13224 genes in the cochlea and midbrain of mice . These findings might contribute to further understanding of pathophysiology and therapy of tinnitus . Influence of phenolic constituents from Yucca schidigera bark on arachidonate metabolism in vitro . Yucca schidigera Roezl . ( Agavaceae ) has been traditionally used to treat a variety of diseases including arthritis and rheumatism . Phenolic constituents isolated from yucca bark , such as resveratrol , trans-3,3',5,5'-tetrahydroxy-4'-methoxystilbene , and the yuccaols , have been shown to possess various activities in vitro , such as antioxidant , radical scavenging , P35228 expression inhibitory , and platelet aggregation inhibitory effects . In the present study , the influence of a phenolic-rich fraction from yucca bark and of its main phenolic constituents on key enzymes of arachidonate metabolism was investigated . The fraction and the pure phenolics were shown to inhibit P23219 , P35354 , and Q06643 4 formation by 5- P28300 in vitro to different extents . The degree of P23219 inhibition was found to be strongly dependent on the substitution pattern of ring B of the stilbenic moiety . The same trend was observed for the P35354 inhibitory potential , which was , however , in general much lower for the yuccaols as compared with resveratrol . DB02709 was also the only compound possessing an Q06643 4 formation inhibitory activity . The inhibitory activity on key enzymes of arachidonate metabolism observed in this study might contribute to the explanation of the anti-inflammatory and antiplatelet effects observed for Y. schidigera and its phenolic constituents . Inhibition of human steroid 5beta-reductase ( P51857 ) by finasteride and structure of the enzyme-inhibitor complex . The Delta(4)-3-ketosteroid functionality is present in nearly all steroid hormones apart from estrogens . The first step in functionalization of the A-ring is mediated in humans by steroid 5alpha- or 5beta-reductase . DB01216 is a mechanism-based inactivator of 5alpha-reductase type 2 with subnanomolar affinity and is widely used as a therapeutic for the treatment of benign prostatic hyperplasia . It is also used for androgen deprivation in hormone-dependent prostate carcinoma , and it has been examined as a chemopreventive agent in prostate cancer . The effect of finasteride on steroid 5beta-reductase ( P51857 ) has not been previously reported . We show that finasteride competitively inhibits P51857 with low micromolar affinity but does not act as a mechanism-based inactivator . The structure of the P51857 .NADP(+)*finasteride complex determined at 1.7 A resolution shows that it is not possible for NADPH to reduce the Delta(1-2)-ene of finasteride because the cofactor and steroid are not proximal to each other . The P01024 -ketone of finasteride accepts hydrogen bonds from the catalytic residues DB00135 -58 and DB00142 -120 in the active site of P51857 , providing an explanation for the competitive inhibition observed . This is the first reported structure of finasteride bound to an enzyme involved in steroid hormone metabolism . Tandutinib ( MLN518 ) reverses multidrug resistance by inhibiting the efflux activity of the multidrug resistance protein 7 ( Q5T3U5 ) . It is well established that DB00171 -binding cassette ( DB01048 ) transporter-mediated multidrug resistance ( MDR ) is one of the major mechanisms that causes resistance to antineoplastic drugs in cancer cells . ABC transporters can significantly decrease the intracellular concentration of antineoplastic drugs by increasing their efflux , thereby lowering their cytotoxic activity . One of these transporters , the multidrug resistance protein 7 ( Q5T3U5 / Q5T3U5 ) , has already been shown to produce resistance to antineoplastic drugs by increasing the efflux of the drugs . In the present study , we investigated whether DB05465 , an P07333 -like tyrosine kinase 3 ( P36888 ) inhibitor , has the potential to reverse Q5T3U5 -mediated MDR . Our results revealed that DB05465 significantly enhanced the sensitivity of Q5T3U5 -transfected HEK293 cells to the 2 established Q5T3U5 substrates , paclitaxel and vincristine , whereas there was less or no effect on the control vector-transfected HEK293 cells . [ ³H ] -paclitaxel accumulation and efflux studies demonstrated that DB05465 increased the intracellular accumulation of [ ³H ] -paclitaxel and inhibited the efflux of [ ³H ] -paclitaxel from P29320 - Q5T3U5 cells . In addition , western blot analysis showed that DB05465 did not significantly affect Q5T3U5 expression . Thus , we conclude that the P36888 inhibitor DB05465 can reverse Q5T3U5 -mediated MDR through inhibition of the drug efflux function and may have potential to be used clinically in combination therapy for cancer patients . Blocking dopamine D2 receptors by haloperidol curtails the beneficial impact of calorie restriction on the metabolic phenotype of high-fat diet induced obese mice . Calorie restriction is the most effective way of expanding life-span and decreasing morbidity . It improves insulin sensitivity and delays the age-related loss of dopamine receptor D(2) ( P14416 ) expression in the brain . Conversely , high-fat feeding is associated with obesity , insulin resistance and a reduced number of P14416 binding sites . We hypothesised that the metabolic benefit of calorie restriction involves the preservation of appropriate P14416 transmission . The food intake of wild-type C57Bl6 male mice was restricted to 60 % of ad lib. intake while they were treated with the P14416 antagonist haloperidol or vehicle using s.c. implanted pellets . Mice with ad lib. access to food receiving vehicle treatment served as controls . All mice received high-fat food throughout the experiment . After 10 weeks , an i.p. glucose tolerance test was performed and , after 12 weeks , a hyperinsulinaemic euglycaemic clamp . Hypothalamic P14416 binding was also determined after 12 weeks of treatment . Calorie-restricted ( CR ) vehicle mice were glucose tolerant and insulin sensitive compared to ad lib . ( AL ) fed vehicle mice . CR mice treated with haloperidol were slightly heavier than vehicle treated CR mice . DB00502 completely abolished the beneficial impact of calorie restriction on glucose tolerance and partly reduced the insulin sensitivity observed in CR vehicle mice . The metabolic differences between AL and CR vehicle mice were not accompanied by alterations in hypothalamic P14416 binding . In conclusion , blocking P14416 curtails the metabolic effects of calorie restriction . Although this suggests that the dopaminergic system could be involved in the metabolic benefits of calorie restriction , restricting access to high-fat food does not increase ( hypothalamic ) P14416 binding capacity , which argues against this inference . Changes in expression of DB01221 receptor subunit mRNA by perinatal exposure to dioxin . Since dioxin and related compounds are suspected of affecting permanently the brain function of offspring of human and experimental animals , effects of perinatal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin ( TCDD ) on the expression of rat DB01221 receptor Q12879 and Q13224 subunit mRNA were examined . The mRNA quantification by competitive RT-PCR clearly revealed that TCDD inhibited Q13224 mRNA expression and enhanced Q12879 mRNA expression in the neocortex and hippocampus on postnatal day ( P01160 ) 49 , whereas these changes in mRNA expression were not found on P01160 5 . The results demonstrate for the first time that the perinatal exposure to TCDD can alter the molecular basis of brain of offspring in adulthood . Molecular identification of the human O75899 : cell surface expression and coupling to adenylyl cyclase in the absence of Q9UBS5 . We have identified a gene encoding a GABAB receptor , the human O75899 , located on chromosome 9q22.1 , that is distinct from the recently reported rat Q9UBS5 . O75899 structurally resembles Q9UBS5 ( 35 % identity ) , having seven transmembrane domains and a large extracellular region , but differs in having a longer carboxy-terminal tail . O75899 is localized to the cell surface in transfected COS cells , and negatively couples to adenylyl cyclase in response to GABA , baclofen , and 3-aminopropyl(methyl)phosphinic acid in CHO cells lacking Q9UBS5 . DB00181 action is inhibited by the GABABR antagonist , 2-hydroxysaclofen . The human O75899 and Q9UBS5 genes are differentially expressed in the nervous system , with the greatest difference being detected in the striatum in which Q9UBS5 but not O75899 mRNA transcripts are detected . O75899 and Q9UBS5 mRNAs are also coexpressed in various brain regions such as the Purkinje cell layer of the cerebellum . Identification of a functional homomeric O75899 coupled to adenylyl cyclase suggests that the complexity of GABAB pharmacological data is at least in part due to the presence of more than one receptor and opens avenues for future research leading to an understanding of metabotropic GABA receptor signal transduction mechanisms . Chronic inhibition of farnesyl pyrophosphate synthase attenuates cardiac hypertrophy and fibrosis in spontaneously hypertensive rats . P14324 ( FPPS ) , an essential enzyme in the mevalonate pathway , was reported to be upregulated in young spontaneously hypertensive rats ( SHR ) when compared with Wistar-Kyoto ( WKY ) rats , and this was accompanied by development of left ventricular hypertrophy . Five-week-old rats were daily gavaged with vehicle or an FPPS inhibitor ( alendronate , 1 or 10 mg/kg ) and blood pressures was monitored by the tail-cuff method every other week . Twelve weeks of alendronate treatment attenuated the left ventricular weight to body weight ratio ( LVW/BW ) , hydroxyproline content , collagen deposition in the interstitia , and gene expression of atrial natriuretic peptide , B-type natriuretic peptide , and procollagen type I/III in the SHR left ventricle , all of which were significantly higher in SHRs than in WKY rats . Furthermore , long-term treatment with an FPPS inhibitor significantly reduced RhoA activation , P29323 phosphorylation , and TGF-beta1 expression in the SHR left ventricle , all of which were upregulated more in SHRs than in WKY rats . In conclusion , chronic treatment with an FPPS inhibitor attenuates the development of cardiac hypertrophy and fibrosis , and the suppression of P27361 /2 phosphorylation and TGF-beta1 expression with inhibition of RhoA activation may be an important mechanism . Pharmacological profile of astemizole-derived compounds at the histamine H1 and H4 receptor -- H1/H4 receptor selectivity . DB00637 , a P35367 antagonist shows high affinity to the histamine H1 receptor but only a moderate affinity to the histamine H4 receptor . This study aims to modify the astemizole to keep high affinity to the histamine H1 receptor and to increase affinity to the histamine H4 receptor . Therefore , 13 astemizole-derived compounds and astemizole-JNJ7777120-derived hybrid compounds were synthesized and pharmacologically characterized at the histamine H1 and H4 receptors . The new compounds show affinity to the histamine H1 receptor in the pK i range from 5.3 to 8.8 , whereas the affinity of these compounds to the histamine H4 receptor was surprisingly rather low ( pK i from 4.4 to 5.6 ) . Three representative compounds were docked into the histamine H1 receptor and molecular dynamic studies were performed to explain the binding mode and the experimental results on a molecular level . Furthermore , taking into account the binding mode of compounds with high affinity to the histamine H4 receptor , a H1/H4-pharmacophore hypothesis was developed . Crystal structure of phenylalanine ammonia lyase : multiple helix dipoles implicated in catalysis . The first three-dimensional structure of phenylalanine ammonia lyase ( Q9P2V4 ) has been determined at 2.1 A resolution for Q9P2V4 from Rhodosporidium toruloides . The enzyme is structurally similar to the mechanistically related histidine ammonia lyase ( P42357 ) , with Q9P2V4 having an additional approximately 160 residues extending from the common fold . We propose that catalysis ( including lowering the pK(a) of nonacidic P01024 of l-phenylalanine for an E1cb mechanism ) is potentially governed by dipole moments of seven alpha helices associated with the Q9P2V4 active site ( six positive poles and one negative pole ) . Cofactor 3,5-dihydro-5-methylidene-4H-imidazol-4-one ( Q9NP71 ) resides atop the positive poles of three helices , for increasing its electrophilicity . The helix dipoles appear fully compatible with a model of phenylalanine docked in the active site of Q9P2V4 having the first covalent bond formed between the amino group of substrate and the methylidene group of Q9NP71 : 12 highly conserved residues ( near the N termini of helices for enhancing function ) are poised to serve roles in substrate recognition , Q9NP71 activation , product separation , proton donation , or polarizing electrons from the phenyl ring of substrate for activation of P01024 ; and a highly conserved DB00117 residue ( near the C terminus of the one helix that directs its negative pole toward the active site to increase the residue 's basicity ) is positioned to act as a general base , abstracting the pro-S hydrogen from P01024 of substrate . A similar mechanism is proposed for P42357 , which has a similar disposition of seven alpha helices and similar active-site residues . The helix dipoles appear incompatible with a proposed mechanism that invokes a carbocation intermediate . Effect of valproic acid through regulation of DB01221 receptor- P29323 signaling in sleep deprivation rats . Although the effect of mood stabilizer valproic acid ( DB00313 ) through multiple signaling pathways has been shown , its therapeutic mechanism is still largely unknown . We investigated the effect of DB00313 ( 200 mg/kg , every 12 h ) in sleep deprivation ( SD ) rats ( 72 h ) , the manic-like animal model , focusing on the N-methyl-D : -aspartic acid ( DB01221 ) receptor and signaling mediators of synaptic plasticity such as extracellular signal-regulated protein kinase ( P29323 ) , DB02527 response element-binding protein ( CREB ) , B cell chronic lymphocytic leukemia/lymphoma 2 ( P10415 ) , and brain-derived neurotrophic factor ( P23560 ) . SD reduced the expression of the Q13224 subunit of the DB01221 receptor in the frontal cortex and hippocampus but did not affect the expression of Q9UHB4 and Q12879 subunits . In comparison , DB00313 inhibited the SD-induced reduction of Q13224 expression in both brain regions . In addition , SD attenuated P29323 phosphorylation in the frontal cortex and hippocampus , whereas DB00313 prevented the attenuation . DB00313 also protected the SD-induced decrease of CREB phosphorylation , P10415 expression , and P23560 expression in the frontal cortex but not in the hippocampus . These results indicate that DB00313 could regulate DB01221 receptor- P29323 signaling in SD rats , preventing the SD-induced decrease of the expression of Q13224 subunit and the activation of P29323 signaling mediators such as P29323 , CREB , P10415 , and P23560 . Access of inhibitory neurosteroids to the DB01221 receptor . BACKGROUND AND PURPOSE : DB01221 receptors are glutamatergic ionotropic receptors involved in excitatory neurotransmission , synaptic plasticity and excitotoxic cell death . Many allosteric modulators can influence the activity of these receptors positively or negatively , with behavioural consequences . 20-Oxo-5β-pregnan-3α-yl sulphate ( pregnanolone sulphate ; PA-6 ) is an endogenous neurosteroid that inhibits DB01221 receptors and is neuroprotective . We tested the hypothesis that the interaction of PA-6 with the plasma membrane is critical for its inhibitory effect at DB01221 receptors . EXPERIMENTAL APPROACH : Electrophysiological recordings and live microscopy were performed on heterologous HEK293 cells expressing Q05586 / Q13224 receptors and cultured rat hippocampal neurons . KEY RESULTS : Our experiments showed that the kinetics of the steroid inhibition were slow and not typical of drug-receptor interaction in an aqueous solution . In addition , the recovery from steroid inhibition was accelerated by β- and γ-cyclodextrin . Values of IC(50) assessed for novel synthetic P01024 analogues of PA-6 differed by more than 30-fold and were positively correlated with the lipophilicity of the PA-6 analogues . Finally , the onset of inhibition induced by P01024 analogues of PA-6 ranged from use-dependent to use-independent . The onset and offset of cell staining by fluorescent analogues of PA-6 were slower than those of steroid-induced inhibition of current responses mediated by DB01221 receptors . CONCLUSION AND IMPLICATIONS : We conclude that steroid accumulation in the plasma membrane is the route by which it accesses a binding site on the DB01221 receptor . Thus , our results provide a possible structural framework for pharmacologically targeting the transmembrane domains of the receptor . P01308 like growth factor-1 selectively regulates the expression of matrix metalloproteinase-2 in malignant H-ras transformed cells . The present study demonstrates alterations in the regulation of matrix metalloproteinase-2 ( P08253 ) expression in response to insulin like growth factor-1 ( DB01277 ) in a H-ras transformed cell line , P01024 , which is capable of metastasis formation . These changes in P08253 expression in response to DB01277 treatment did not occur in either non-transformed parental 10 T 1/2 cells or in H-ras transformed cells ( Q13224 cells ) which are capable of benign tumour formation . DB01277 treatment of P01024 cells resulted in increased expression of P08253 gelatinolytic activity and increased expression of P08253 mRNA levels . The DB01277 mediated alterations in P08253 mRNA levels were dependent upon de novo protein synthesis and independent of transcriptional events , but dependent upon post-transcriptional regulatory events . Most notably , DB01277 can regulate P08253 mRNA expression in P01024 cells through a mechanism involving P08253 message stabilization . This study demonstrates aspects of the temporal regulatory mechanisms of P08253 expression in response to insulin-like growth factor-1 in a H-ras transformed fibrosarcoma cell line capable of metastasis formation and thereby , provides further insight into the altered growth regulatory program associated with H-ras mediated cellular transformation and malignant progression . Matrix proteinase inhibition by DB05387 , a multifunctional antiangiogenic compound . BACKGROUND : Matrix metalloproteinases ( MMPs ) play an important role in tissue remodeling under normal physiological and pathological conditions and are thus attractive targets for both diagnostic and therapeutic purposes . Here , we examined the effect of DB05387 , an orally bioavailable standardized extract made of cartilage that shows significant antiangiogenic and antimetastatic properties in vivo , on the activity of various members of the MMP family . MATERIALS AND METHODS : The effect of DB05387 on the activity of MMPs was assessed by fluorimetric assays and by substrate gel zymography . RESULTS : DB05387 markedly inhibits the gelatinolytic activity of P08253 and to a lesser extent those of P03956 , P09237 , P14780 and P45452 . DB05387 also inhibited the elastinolytic activities of P08253 and P14780 as well as P39900 ( metalloelastase ) , porcine pancreatic elastase ( PPE ) , and human leukocyte elastase ( P08246 ) . Western blot analysis revealed the presence within DB05387 of immunoreactive P01033 -like proteins , suggesting that these proteins may be at least partly responsible for the observed MMP inhibition . CONCLUSIONS : Taken together , these results demonstrate that DB05387 contains P01033 -like proteins that could be responsible for the specific inhibition of MMPs . Given the recent studies suggesting the presence within this compound of specific inhibitor(s) of endothelial cell proliferation , DB05387 appears as a pleotropic agent able to interfere with several biochemical steps leading to angiogenesis and to other physiopathological conditions . Since DB05387 is currently under Phase III clinical investigations , these findings are also of considerable importance for our understanding of its anticancer properties .

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interacts_with DB00563?

multiple_choice

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 . The blockage of Ras/ P29323 pathway augments the sensitivity of SphK1 inhibitor P12755 II in human hepatoma HepG2 cells . The treatment of hepatocellular carcinoma ( HCC ) remains a challenge and the future of cancer therapy will incorporate rational combinations directed to molecular targets that cooperate to drive critical pro-survival signaling . Q9NYA1 ( SphK1 ) has been shown to regulate various processes important for cancer progression . Given the up-regulated expression of SphK1 in response to the silence of N-ras and other interactions between Ras/ P29323 and SphK1 , it was speculated that combined inhibition of Ras/ P29323 and SphK1 would create enhanced antitumor effects . Experimental results showed that dual blockage of N-ras/ P29323 and SphK1 resulted in enhanced growth inhibitions in human hepatoma cells . Similarly , Q02750 /2 Inhibitor U0126 potentiated P12755 II-induced apoptosis in hepatoma HepG2 cells , consistently with the further attenuation of Akt/ P29323 /NF-κB signaling pathway . It was also shown that the combination of P12755 II and U0126 further attenuated the migration of hepatoma HepG2 cells via Q05397 /MLC-2 signaling pathway . Taken together , the dual inhibition of SphK1 and Ras/ P29323 pathway resulted in enhanced effects , which might be an effective therapeutic approach for the treatment of HCC . Selective effect of INGAP-PP upon mouse embryonic stem cell differentiation toward islet cells . We evaluated the effect of islet neogenesis-associated protein pentadecapeptide ( INGAP-PP ) upon islet beta- and non-beta cell differentiation from mouse embryonic stem ( mES ) cells . ES-D3 cell lines were cultured following Lumelsky 's protocol with or without INGAP-PP ( 5 microg/ml ) at different stages . Gene expression was quantified using qPCR. mES cells were fixed and immunostained using anti insulin- , somatostatin- , glucagon- , Pdx-1- , Ngn-3- , Nkx-6.1 and P09936 specific antibodies . P12004 was used to measure replication rate . Bcl(2) ( immunostaining ) and caspase-3 ( enzyme activity and gene expression ) were determined as apoptosis markers . INGAP-PP increased P10997 , Glut-2 , Kir-6.2 , Q09428 -1 and insulin gene expression , and the percentage of insulin-immunostained cells . Conversely , INGAP-PP reduced significantly glucagon and somatostatin gene expression and immunopositivity . While nestin gene expression was not affected , there was a significant reduction in the percentage of P09936 -immunostained cells . Pdx-1 gene expression increased by 115 % in INGAP-PP treated cells , as well as the percentage of Pdx-1 , Ngn-3 and Nkx-6.1 immunopositive cells . Neither caspase-3 ( expression and activity ) nor Bcl(2) positively immunostained cells were affected by INGAP-PP . Accordingly , INGAP-PP would promote stem cell differentiation into a beta-like cell phenotype , simultaneously decreasing its differentiation toward non-beta-cell precursors . Therefore , INGAP-PP would be potentially useful to obtain beta-cells from stem cells for replacement therapy . DB00563 in rheumatoid arthritis : studies with animal models . The present studies have shown that low doses of methotrexate can suppress the inflammation and joint destruction associated with animal models of arthritis . The antiinflammatory effects of methotrexate are probably related to its inhibitory effect on chemotaxis . At the low doses used , methotrexate does not induce systemic immunosuppression . In methotrexate-treated rats , an improvement in P60568 synthesis is observed and increases in P60568 levels are expected to improve cell mediated immunity . Suppressor cells appear to be very sensitive to methotrexate . Macrophage function is modulated by methotrexate . All of these effects including the effects on joint destruction are probably due to inhibition of P00374 activity of critical cells that are involved in the pathogenesis of rat arthritis induced either by adjuvant or by streptococcal cell walls . Some of these effects have been extended to human arthritis but additional studies are required to understand how low dose methotrexate exerts its beneficial effects in humans . Structures of murine carbonic anhydrase IV and human carbonic anhydrase II complexed with brinzolamide : molecular basis of isozyme-drug discrimination . P22748 ( CAIV ) is a membrane-associated enzyme anchored to plasma membrane surfaces by a phosphatidylinositol glycan linkage . We have determined the 2.8-angstroms resolution crystal structure of a truncated , soluble form of recombinant murine CAIV . We have also determined the structure of its complex with a drug used for glaucoma therapy , the sulfonamide inhibitor brinzolamide ( DB01194 ) . The overall structure of murine CAIV is generally similar to that of human CAIV ; however , some local structural differences are found in the active site resulting from amino acid sequence differences in the " 130 's segment " and the residue-63 loop ( these may affect the nearby catalytic proton shuttle , DB00117 -64 ) . Similar to human CAIV , the C-terminus of murine CAIV is surrounded by a substantial electropositive surface potential that may stabilize the interaction with the phospholipid membrane . Binding interactions observed for brinzolamide rationalize the generally weaker affinity of inhibitors used in glaucoma therapy toward CAIV compared with CAII . Recent advances in autoimmune pancreatitis : concept , diagnosis , and pathogenesis . Recent advances support the concept of autoimmune pancreatitis ( AIP ) as a unique systemic disease , because it shows occasional extrapancreatic lesions such as sclerosing cholangitis , sclerosing sialoadenitis , and retroperitoneal fibrosis , pathological features similar to those of fibrosis , and abundant infiltration of IgG4-positive plasma cells , and it is steroid responsive . Based on these findings , several diagnostic criteria have been proposed . Although AIP is accepted worldwide as a unique clinical entity , its pathogenetic mechanism remains unclear . To clarify its pathogenesis , its genetic background , humoral immunity , candidate target antigens including self-antigens and molecular mimicry by microbes , and cellular immunity including regulatory T cells , the complement system , and experimental models are reviewed . On the basis of this review , we hypothesize that the pathogenesis of AIP involves a biphasic mechanism consisting of " induction " and " progression . " In the early stage , the initial response to self-antigens [ lactoferrin , carbonic anhydrase ( CA ) -II , P22748 , pancreatic secretory trypsin inhibitor , and alpha-fodrin ] and molecular mimicry ( Helicobacter pylori ) are induced by decreased naïve regulatory T cells ( Tregs ) , and T-helper ( Th ) 1 cells release proinflammatory cytokines [ interferon-gamma , interleukin ( IL ) -1beta , P60568 , and tumor necrosis factor alpha ] . In the chronic stage , progression is supported by increased memory Tregs and Th2 immune responses . The classical complement system pathway may be activated by the IgG1 immune complex . As Tregs seem to play an important role in progression as well as in induction of the disease , further studies are necessary to clarify the pathogenesis of AIP . Flow cytometric analysis of mammalian glial cultures treated with methotrexate . DB00563 ( MTX ) is an antineoplastic drug that acts by competitive inhibition of the enzyme dihydrofolate reductase ( P00374 ) . MTX treatment of cultured cell lines leads to the emergence of resistant cell populations . Studies using stepwise selection procedures have demonstrated that MTX resistance conferred by overproduction of P00374 can be caused by P00374 gene amplification . We examined the effect of MTX on cells whose origin more closely approximates the in vivo condition by developing a culture system using dissociated brain tissue from 17-19 day old mouse embryos . At the first passage , cultures were divided into control and MTX groups . Cells were treated with the same or successively higher concentrations of MTX at each passage over a 3-4 month period . The first passage eliminated neurons and left a glial culture comprised of approximately 90 % astrocytes . We used the Fluorescence Activated Cell Sorter in conjunction with fluorescent dyes to measure P00374 content , DNA content , size , and viability of glial cells following MTX treatment . MTX-treated cells divided but grew more slowly and were larger than untreated cells . Stepwise selection in 30/60/90 nM or 60/120 nM MTX resulted in significant two- to threefold increases in fluorescence , and hence P00374 levels . Slot hybridizations assays demonstrated a threefold increase in P00374 gene copy number in the DNA from the 30/60/90 cultures . Thus , our findings were consistent with the results obtained from somatic cell lines , and lend support to the hypothesis that gene amplification may be a common mechanism for the acquisition of resistance in many types of cells . They also indicate that glial cells may be a specific target for cytotoxic effects of MTX on the central nervous system . [ Regulation of P04271 expression during long term potentiation ] . In this study , contributions of intracellular regulatory cascades in the induction of P04271 expression in rat hippocampal P00915 area during long term posttetanic potentiation ( LTP ) were estimated . The activation of transcription factor p53 ( positive regulator of P04271 transcription ) by nutlin-3 increased the basal content of P04271 mRNA up to 151 % of the control level , which was significantly lower than its content in tetanized slices ( 280 % ) . Therefore , p53 seems to be not unique transcription factor upregulating P04271 expression during LTP . The inhibitor of Ca2+/calmodulin-dependent kinases ( CaMKs ) KN-93 fully blocked the increase of P04271 mRNA after tetanization , while KN-92 ( inactive analogue of KN-93 ) was ineffective . The inhibitor of CaMKII and receptor tyrosine kinases DB02152 essentially suppressed P04271 expression during LTP , the inhibition of MAPK p38 or P51812 moderately decreased , and the inhibition of Q02750 did not influence P04271 mRNA content . Thus , CaMKs play a key role in the induction of P04271 expression during LTP . Transcript and protein profiling identifies signaling , growth arrest , apoptosis , and NF-κB survival signatures following P01148 receptor activation . P01148 significantly inhibits proliferation of a proportion of cancer cell lines by activating P01148 receptor ( P30968 ) -G protein signaling . Therefore , manipulation of P30968 signaling may have an under-utilized role in treating certain breast and ovarian cancers . However , the precise signaling pathways necessary for the effect and the features of cellular responses remain poorly defined . We used transcriptomic and proteomic profiling approaches to characterize the effects of P30968 activation in sensitive cells ( HEK293- P30968 , SCL60 ) in vitro and in vivo , compared to unresponsive HEK293 . Analyses of gene expression demonstrated a dynamic response to the P01148 superagonist DB06825 . Early and mid-phase changes ( 0.5-1.0 h ) comprised mainly transcription factors . Later changes ( 8-24 h ) included a P01148 target gene , P01215 , and up- or downregulation of transcripts encoding signaling and cell division machinery . Pathway analysis identified altered MAPK and cell cycle pathways , consistent with occurrence of G(2)/M arrest and apoptosis . Nuclear factor kappa B ( NF-κB ) pathway gene transcripts were differentially expressed between control and DB06825 -treated SCL60 cultures . Reverse-phase protein and phospho-proteomic array analyses profiled responses in cultured cells and SCL60 xenografts in vivo during DB06825 anti-proliferation . Increased phosphorylated NF-κB ( p65 ) occurred in SCL60 in vitro , and p-NF-κB and IκBε were higher in treated xenografts than controls after 4 days DB06825 . NF-κB inhibition enhanced the anti-proliferative effect of DB06825 in SCL60 cultures . This study reveals details of pathways interacting with intense P30968 signaling , identifies potential anti-proliferative target genes , and implicates the NF-κB survival pathway as a node for enhancing P01148 agonist-induced anti-proliferation . [ Meloxicam ( DB00814 ) : a review of its pharmacological and clinical profile ] . Meloxicam ( DB00814 ) is a new nonsteroidal anti-inflammatory drug ( NSAID ) derived from enolic acid , exhibiting selectivity for cyclooxygenase ( P36551 ) -2 over P23219 . Meloxicam has shown potent anti-inflammatory and analgesic activity together with low gastrointestinal toxicity in animal models . It is a potent inhibitor not only of acute exudation in adjuvant arthritis in the rat , but also of bone and cartilage destruction . The therapeutic range of meloxicam in the rat , with regard to inhibition of adjuvant arthritis , was several times greater than that of other NSAIDs . Meloxicam in therapeutic doses was found to have no effect on bleeding time or platelet aggregation in healthy volunteers . In clinical studies , meloxicam has shown reliable efficacy against rheumatoid arthritis , osteoarthritis , lumbago ( low back pain ) , scapulohumeral periarthritis , and neck-shoulder-arm syndrome with low gastrointestinal toxicity . The receptor tyrosine kinase inhibitor amuvatinib ( DB05216 ) sensitizes tumor cells to radio- and chemo-therapies in part by inhibiting homologous recombination . BACKGROUND AND PURPOSE : Q06609 is a key protein involved in homologous recombination ( HR ) and a potential target for radiation- and chemotherapies . Amuvatinib ( formerly known as DB05216 ) is a novel receptor tyrosine kinase inhibitor that targets c- P10721 and PDGFRα and can sensitize tumor cells to ionizing radiation ( IR ) . Here , we studied amuvatinib mechanism on Q06609 and functional HR . MATERIALS AND METHODS : Protein and RNA analyses , direct repeat green fluorescent protein ( DR-GFP ) assay and polysomal fractioning were used to measure HR efficiency and global translation in amuvatinib-treated H1299 lung carcinoma cells . Synergy of amuvatinib with IR or mitomycin c ( DB00305 ) was assessed by clonogenic survival assay . RESULTS : Amuvaninib inhibited Q06609 protein expression and HR . This was associated with reduced ribosomal protein S6 phosphorylation and inhibition of global translation . Amuvatinib sensitized cells to IR and DB00305 , agents that are selectively toxic to HR-deficient cells . CONCLUSIONS : Amuvatinib is a promising agent that may be used to decrease tumor cell resistance . Our work suggests that this is associated with decreased Q06609 expression and function and supports the further study of amuvatinib in combination with chemotherapy and radiotherapy . Reduced folate carrier and dihydrofolate reductase expression in acute lymphocytic leukemia may predict outcome : a Children 's Cancer Group Study . PURPOSE : DB00563 is a major component of current treatment regimens for children with acute lymphocytic leukemia ( ALL ) . Potential mechanisms of methotrexate resistance include impaired drug uptake , decreased drug retention , and dihydrofolate reductase ( P00374 ) amplification . The purpose of this study was to assess whether reduced folate carrier ( P41440 ) and P00374 expression in untreated leukemic blasts correlated with outcome . METHODS : Quantitative real-time RT-PCR was used to measure P41440 and P00374 mRNA expression in leukemic blasts from 40 newly diagnosed patients with ALL obtained in a blinded fashion from Children 's Cancer Group studies . RESULTS : Low P41440 expression at diagnosis correlated significantly with an unfavorable event free survival . Surprisingly , low , not high , P00374 expression correlated significantly with an unfavorable event-free survival . Proliferative cell nuclear antigen ( P12004 ) expression demonstrated a weak inverse relationship between sample P12004 and P00374 or P41440 expression , suggesting that P00374 and P41440 expression may be markers for factors other than drug resistance . CONCLUSIONS : These results suggest that impaired transport may be an important mechanism of intrinsic methotrexate resistance in ALL , and P00374 expression also may be an important prognostic factor in ALL . Additional studies are necessary to clarify the mechanism for the correlation of low P00374 expression with poor outcome . DB05332 administration shows reduced megakaryocyte response-capacity and increased myelofibrosis in a mouse model of P35579 -RD . Macrothrombocytopenia in P35579 -related disease ( P35579 -RD ) results from defects in nonmuscular myosin-IIA function . P40238 agonists ( eltrombopag ; romiplostim ) seem to improve hemostasis , but little is known about their biologic effects in P35579 -RD . We administered romiplostim to Myh9(-/-) mice ( 100 μg/kg , every 3 days , during 1 month ) . MKs increased to similar numbers in Myh9(-/-) and wild-type ( WT ) mice ( with an increase in immature MKs ) , but Myh9(-/-) platelet count response was much less ( 2.5-fold vs 8-fold increase ) . A strong increase in MK nuclei emboli in the lung , in WT and Myh9(-/-) mice , indicates increased transmigration of MKs from the BM . Prolonged ( but not acute ) treatment with romiplostim decreased expression of GPIb-IX-V complex and Q9HCN6 , but not of GPIIbIIIa , and bleeding time increased in WT mice . Microcirculation was not altered by the increased number of large platelets in any of the assessed organs , but in Myh9(-/-) mice a much stronger increase in BM reticulin fibers was present after 4 weeks of romiplostim treatment vs WT mice . These data further encourage short-term use of thrombopoietic agents in patients with P35579 -RDs ; however , myelofibrosis has to be considered as a potential severe adverse effect during longer treatment . Reduction of GPIbIX/ Q9HCN6 expression by romiplostim requires further studies . DB00563 and cytarabine inhibit progression of human lymphoma in NOD/SCID mice carrying a mutant dihydrofolate reductase and cytidine deaminase fusion gene . An SFG-based retroviral bicistronic vector containing a double-mutant dihydrofolate reductase-cytidine deaminase fusion cDNA ( F/S P00374 -CD ) with IRES-eGFP confers resistance to both methotrexate ( MTX ) and cytarabine ( ara-C ) . Two weeks after transplantation with marrow transduced with either a fusion or a control gene ( eGFP-IRES-NeoR ) , human lymphoma ( P12755 -DLCL-1 ) cells were injected sc into the flanks of nonobese diabetic/severe combined immune deficiency mice . In mock-transplanted mice , maximal tolerated dose ( MTD ) of posttransplant MTX/ara-C ( 15/10 mg/kg/day , x3 ) was unable to control tumor growth . Transfer of the fusion gene allowed doses of MTX/ara-C ( 25/15 mg/kg/day , x4 ) twofold higher than the MTD to be tolerated . The tumor burden defined the efficiency of posttransplant chemotherapy ; early treatment , 48 h after tumor inoculation , provided tumor-free survival , while starting treatment after having palpable tumor growth ( 7 days ) delayed tumor growth a median time of 28 days . In addition , the early treated group had higher gene expression in peripheral blood and marrow cells than the late treated group ( P < 0.05 ) , suggesting that early treatment allowed for enrichment of transduced marrow progenitors . These results encourage clinical studies using this retroviral fusion gene construct . Use of fuzzy neural networks in modeling relationships of HPV infection with apoptotic and proliferation markers in potentially malignant oral lesions . To evaluate in oral leukoplakia the relationship between HPV infection and markers of apoptosis ( bcl-2 , survivin ) and proliferation ( P12004 ) , also conditionally to age , gender , smoking and drinking habits of patients , by means of Fuzzy neural networks ( FNN ) system 21 cases of oral leukopakia , clinically and histologically diagnosed , were examined for HPV DNA presence , bcl-2 , survivin and P12004 expression . HPV DNA was investigated in exfoliated oral mucosa cells by nested PCR ( nPCR : MY09-MY11/ P40197 - Q9HCN6 ) , and the HPV genotype determined by direct DNA sequencing . All markers were investigated by means of standardised immunohistochemistry procedure . Data were analysed by chi-square test , crude OR and the 95 % CI ; in blindness , FNN was applied . HPV DNA was found in 8/21 OL ( 38.1 % ) ; survivin , P12004 , and tobacco smoking were associated in univariate analysis ( p = 0.04 ) with HPV DNA status . HPV-18 was the most frequently detected genotype ( 6/8 ) , followed by HPV-16 ( 2/8 ) . FNN revealed that survivin and P12004 , both being expressed in all of OL HPV+ve , were associated with HPV infection . In conclusion , the FNN allowed to hypothesise a model of specific variables associated to HPV infection in OL . The relevance of survivin and P12004 suggest that they may be involved in HPV-mediated deregulation of epithelial maturation and , conversely , that HPV may have a role in the expression level of these two markers . FNN system seems to be an effective tool in the analysis of correlates of OL and HPV infection . An androgen receptor gene mutation ( A645D ) in a boy with a normal phenotype . Over 100 mutations have so far been published in the androgen receptor gene ( AR ) in patients with different degrees of undervirilisation . The AR gene consists of 8 exons , exons 2-3 code for the DNA binding domain and exons 4-8 ( codon 628-919 ) for the steroid binding domain ( codon 667-919 ) . Only four mutations have been published in the 5' end of exon 4 , codons 628-667 , that is in two P22234 cases and two prostate tumours . In a set of phenotypically normal controls we observed one mutation in exon 4 , codon 645 , in a 15 year old boy . He has a history of Wilms ' tumour but no undervirilisation and without a family history with intersex . Exon 4 was amplified from constitutional DNA and subject to DGGE with 35-65 % of denaturants . Aberrant fragments were subject to a new PCR and direct sequencing was performed . This mutation changed Ala645 --> DB00128 due to change of GCT --> Q6IB77 This was not detected in 108 normal chromosomes . Interestingly , the same mutation was recently reported in one P22234 case . This phenotypic discrepancy calls for functional studies of this region . This is the first case of a normal phenotype with an amino acid alteration in the AR gene . [ Pharmacological and clinical profile of mitiglinide calcium hydrate ( Glufast ) , a new insulinotropic agent with rapid onset ] . DB01252 calcium hydrate ( mitiglinide , Glufast ) is a new insulinotropic agent of the glinide class with rapid onset . DB01252 is thought to stimulate insulin secretion by closing the DB00171 -sensitive K(+) ( K( DB00171 ) ) channels in pancreatic beta-cells , and its early insulin release and short duration of action would be effective in improving postprandial hyperglycemia . In studies of various cloned K( DB00171 ) channels , mitiglinide shows a higher selectivity for the beta-cell type of Q09428 /Kir6.2 than the cardiac and smooth muscle types of K( DB00171 ) channels in comparison with glibenclamide and glimepiride . In vitro and in vivo studies demonstrated the insulinotropic effect of mitiglinide is more potent than that of nateglinide , and mitiglinide surpassed in controlling postprandial hyperglycemia in normal and diabetic animals . In clinical trials , treatment with mitiglinide provided lasting improvement of postprandial hyperglycemia in Type 2 diabetic patients and decreased the fasting plasma glucose levels and HbA(1C) values . The incidence of adverse events related to mitiglinide were nearly equivalent to placebo ; in particular there was no difference with the frequency of hypoglycemia . The results from these studies indicated that mitiglinide could be expected to possess good therapeutic features of being effective in reducing postprandial glucose excursions in the early stage of Type 2 diabetes and less incidence of events suggestive of hypoglycemia . Copy number analysis of 24 oncogenes : O15151 identified as a putative marker for low recurrence risk in non muscle invasive bladder cancer . Patients with non-muscle invasive bladder cancer ( NMIBC ) generally have a high risk of relapsing locally after primary tumor resection . The search for new predictive markers of local recurrence thus represents an important goal for the management of this disease . We studied the copy number variations ( CNVs ) of 24 oncogenes ( O15151 , P04198 , Q9UM73 , P16234 , P10721 , P35968 , P00374 , P00533 , MET , SMO , P11362 , MYC , P00519 , P07949 , P24385 , P30279 , P11802 , Q00987 , Q96GD4 , P04626 , P11388 , O14965 , AR and P15056 ) using multiplex ligation probe amplification technique to verify their role as predictive markers of recurrence . DB03843 -fixed paraffin-embedded tissue samples from 43 patients who underwent transurethral resection of the bladder ( TURB ) were used ; 23 patients had relapsed and 20 were disease-free after 5 years . Amplification frequencies were analyzed for all genes and O15151 was the only gene that showed significantly higher amplification in non recurrent patients than in recurrent ones ( 0.65 vs. 0.3 ; Fisher 's test p=0.023 ) . Recurrence-free survival analysis confirmed the predictive role of O15151 ( log-rank test p=0.041 ) . Our preliminary results indicate a putative role for the O15151 gene in predicting local recurrence of bladder cancer . Confirmation of this hypothesis is needed in a larger cohort of NMIBC patients . Effect of luteinizing hormone-releasing hormone ( P01148 ) analogue treatment on a cytokine profile in prostate cancer patients . The aim of the study was to test serum concentrations of the chosen cytokines in patients with prostate cancer ( PCa ) treated with an luteinizing hormone-releasing hormone ( P01148 ) analogue . We tested interleukin ( IL ) -2 , P22301 , tumor necrosis factor ( P01375 ) -alpha , interferon ( P27352 ) -gamma in blood at three time points ; I - before the injection , II - 10 days and III - 20 days after the injection in 14 men with PCa . Patients had one depot injection of the P01148 analogue monthly . The cytokine concentrations in serum samples were determined by ELISA method . Prostate specific antigen ( PSA ) level was examined before and after six months of the P01148 analogue treatment . After six months of the therapy , we observed normalization of serum PSA value from 16.48 ng/ml to 1.45 ng/ml . P01148 analogue injection resulted in a significant drop of the P60568 concentration , and the value gradually returned to normal in the next 20 days . P22301 concentration transiently increased and then was down-regulated . Serum P01375 and P27352 -gamma concentrations in PCa patients were significantly lower compared to controls and were not affected by the treatment . P01148 analogue treatment in PCa patients modulates concentrations of the chosen cytokines which may result both in antitumor and a transient immunosuppressive effect . P12004 connects DNA replication to epigenetic inheritance in yeast . Formation of a heterochromatin-like structure results in transcriptional silencing at the HM mating-type loci and telomeres in Saccharomyces cerevisiae . Once formed , such epigenetically determined structures are inherited for many mitotic divisions . Here we show that mutations in the proliferating cell nuclear antigen ( P12004 ) , an essential component at the DNA replication fork , reduced repression of genes near a telomere and at the silent mating-typelocus , P22736 . The pol30-8 mutant displayed coexistence of both repressed ( pink ) and de-repressed ( white ) cells within a single colony when assayed with the P22234 gene inserted at P22736 . Unlike pol30-8 , the pol30-6 and pol30-79 mutants partially reduced gene silencing at telomeres and the P22736 and synergistically decreased silencing in cells lacking chromatin assembly factor 1 ( Q9UIV1 ) . All silencing defective mutants showed reduced binding to Q9UIV1 in vitro and altered chromatin association of the Q9UIV1 large subunit in vivo . Thus , P12004 participates in inheritance of both DNA and epigenetic chromatin structures during the S phase of the cell cycle , the latter by at least two mechanisms . Current situation of DB01269 , DB05101 , Nimotuzumab and Zalutumumab . P00533 overexpression usually correlates with a more advanced disease stage , a poorer prognosis and a worse chemotherapy response . P00533 expression increase has been observed in many tumours . For all the aforementioned reasons , P00533 inhibition can be considered an attractive approach in cancer treatment . One strategy has been receptor inhibition of extracellular domain using monoclonal antibodies . Cetuximab is the most developed one and there is plenty information on the literature about its current status . In this review we focus on other P00533 monoclonal antibodies under clinical development . The more developed one is DB01269 . Its clinical development is taking place very quickly and it has mainly been studied in colorectal cancer showing promising results . There are also other interesting drugs such as DB05101 , Nimotuzumab and Zalutumumab .

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The regulation of rotenone-induced inflammatory factor production by DB00171 -sensitive potassium channel expressed in BV-2 cells . Our previous studies have demonstrated that activating DB00171 -sensitive potassium channel ( K( DB00171 ) channel ) , not only improved Parkinsonian behavior and neurochemical symptoms , but also reduced P35228 activity and mRNA levels in striatum and nigra of rotenone rat model of Parkinson 's disease ( PD ) . In this study , it was first shown that the subunits of K( DB00171 ) channels are expressed in BV-2 cells , and then it was investigated whether K( DB00171 ) channel was involved in regulating inflammatory factor production from BV-2 cells activated by rotenone . It was found that K( DB00171 ) channel was expressed in BV-2 cell and formed by the combination of Kir 6.1 and Q09428 2A/2B . K( DB00171 ) channel openers ( KCOs ) including pinacidil , diazoxide and iptakalim ( Ipt ) exerted beneficial effects on rotenone-induced morphological alterations of BV-2 cells , decreased tumor necrosis factor alpha ( P01375 ) production and the expression and activity of inducible isoform of nitric oxide synthase ( P35228 ) . Either glibenclamide or 5-hydroxydecanoate acid ( a selective mitochondrial K( DB00171 ) channel blocker ) could abolish the effects of KCOs , suggesting that K( DB00171 ) channels , especially mitochondrial DB00171 -sensitive potassium channels ( mitoK( DB00171 ) channels ) , played a crucial role in preventing the activation of BV-2 cells , and subsequently the production of a variety of proinflammatory factors . Therefore , activation of K( DB00171 ) channel might be a new therapeutic strategy for treating neuroinflammatory and neurodegenerative disorders . Effects of neuropeptides on human lung fibroblast proliferation and chemotaxis . An increase in subepithelial mesenchymal cells and associated connective tissue is a feature of bronchial asthma . We determined whether neuropeptides could modulate fibroblast activity , particularly with respect to proliferation and chemotaxis . Human lung fibroblasts were cultured with neurokinin A ( P20366 ) , DB05875 ( SP ) , vasoactive intestinal peptide ( P01282 ) , and calcitonin-gene-related peptide ( P80511 ) . After 48 h , fibroblast proliferation was measured by a colorimetric assay based on the uptake and subsequent release of methylene blue . The chemotactic response to neuropeptides was determined with the use of a modified Boyden chamber . Both P20366 and SP ( 10(-7)-10(-4) M ) stimulated human lung fibroblast proliferation in Q03591 and IMR-90 fibroblasts . P01282 and P80511 had no effect on fibroblast proliferation . P20366 alone stimulated fibroblast chemotaxis maximally at 10(-10) M . P08473 ( NEP ) activity of 0.52 and 5.2 pmol/10(6) cells was assayed in IMR-90 and Hs68 fibroblasts , respectively . DB02557 ( 5 x 10(-6)-10(-5) M ) , an NEP inhibitor , enhanced fibroblast proliferation in a dose-dependent manner . Thus neuropeptides have the potential to cause activation of mesenchymal cells , and neuropeptide release may contribute to the structural abnormalities observed in asthmatic airways . C. elegans vulval development as a model system to study the cancer biology of P00533 signaling . Molecular genetic studies of C. elegans vulval development have helped to define an evolutionarily conserved signaling pathway from an P01133 -like ligand through P01133 -receptor , Ras and Q96HU1 kinase to the nucleus . Further studies have identified novel positive regulators such as Q8IVT5 -1 and Q09428 -8/ Q5T124 -2 and negative regulators such as cbl/SLI-1 . The many negative regulatory proteins might serve to prevent inappropriate signaling , and thus are analogous to tumor suppressor genes . Guanylyl cyclase / atrial natriuretic peptide receptor-A : role in the pathophysiology of cardiovascular regulation . Atrial natriuretic factor ( ANF ) , also known as atrial natriuretic peptide ( P01160 ) , is an endogenous and potent hypotensive hormone that elicits natriuretic , diuretic , vasorelaxant , and anti-proliferative effects , which are important in the control of blood pressure and cardiovascular events . One principal locus involved in the regulatory action of P01160 and brain natriuretic peptide ( DB04899 ) is guanylyl cyclase / natriuretic peptide receptor-A ( P16066 /NPRA ) . Studies on P01160 , DB04899 , and their receptor , P16066 /NPRA , have greatly increased our knowledge of the control of hypertension and cardiovascular disorders . Cellular , biochemical , and molecular studies have helped to delineate the receptor function and signaling mechanisms of NPRA . Gene-targeted and transgenic mouse models have advanced our understanding of the importance of P01160 , DB04899 , and P16066 /NPRA in disease states at the molecular level . Importantly , P01160 and DB04899 are used as critical markers of cardiac events ; however , their therapeutic potentials for the diagnosis and treatment of hypertension , heart failure , and stroke have just begun to be realized . We are now just at the initial stage of molecular therapeutics and pharmacogenomic advancement of the natriuretic peptides . More investigations should be undertaken and ongoing ones be extended in this important field . Heterozygous missense mutations in the insulin gene are linked to permanent diabetes appearing in the neonatal period or in early infancy : a report from the French ND ( Neonatal Diabetes ) Study Group . OBJECTIVE : Permanent neonatal diabetes ( P01160 ) is defined by chronic hyperglycemia due to severe nonautoimmune insulin deficiency diagnosed in the first months of life . Several genes , including Q14654 and Q09428 , which encode the two subunits of the DB00171 -sensitive K(+) channel ( K( DB00171 ) channel ) can cause P01160 . Mutations in the insulin ( P01308 ) gene have been recently described in families with neonatal diabetes . Our study aimed to investigate the genetic anomalies and clinical heterogeneity in P01160 patients who are negative for a K( DB00171 ) channel mutation . RESEARCH DESIGN AND METHODS : We screened the P01308 gene by direct sequencing in 38 P01160 patients and in one child with nonautoimmune early-infancy diabetes , where no mutation in GCK , Q14654 , and Q09428 was identified . A detailed clinical phenotyping of the patients was carried out to specify the diabetes features in those found with an P01308 mutation . RESULTS : We identified three missense mutations in the P01308 gene in four probands . Two of four mutations were inherited in a dominant manner , and the familial description evidenced a marked variability in age of diagnosis and disease progression . In our cohort , the P01308 mutations may represent approximately 10 % of all permanent neonatal diabetes cases , having a later presentation of diabetes and no associated symptoms compared with cases with K( DB00171 ) channel mutations . CONCLUSIONS ; Heterozygous P01308 gene mutations can cause isolated permanent early-infancy diabetes and should be assessed in neonatal as well as in childhood diabetes appearing like type 1 , when autoimmune markers are absent . New pharmacogenomic strategies may be applicable , since residual beta-cell function is still present in some patients . Regulatory regions of growth-related genes can activate an exogenous gene of the alpha-fetoprotein promoter to a comparable degree in human hepatocellular carcinoma cells . We examined the transcriptional activation by the regulatory regions of the midkine ( MK ) , survivin ( Q09428 ) , cyclooxygenase-2 ( P35354 ) , telomerase reverse transcriptase ( O14746 ) and alpha-fetoprotein ( AFP ) genes in human hepatocellular carcinoma cells . Luciferase assays showed that the Q09428 regulatory region exhibited the greatest activity and that the MK regulatory region activated the reporter gene better than the enhancer-linked AFP promoter even in high-AFP-producing cells . The P35354 and O14746 regulatory regions also activated the reporter gene better than the AFP enhancer/promoter in intermediate-AFP-producing cells . Combination of the regulatory regions arranged in tandem modulated their transcriptional activities , depending on the arrangement of the promoters and cells examined . These data suggested that the regulatory regions of the growth-related genes could be useful to activate a therapeutic gene in hepatocellular carcinoma cells irrespective of the amounts of AFP production but combinatory use of the promoter regions could not always contribute to enhanced activity . DB05822 , a nitric oxide-releasing aspirin derivative , exhibits a significant antiproliferative effect and alters cell cycle progression in human colon adenocarcinoma cell lines . DB00435 -releasing non-steroidal antiinflammatory drugs ( NO-NSAIDs ) are safer than NSAIDs due to their ability to reduce gastric toxicity . We assessed the cytotoxic activity of a new aspirin derivative , DB05822 , after different exposure schedules , in three human colon adenocarcinoma cell lines . All the lines were positive for P23219 protein and mRNA , as evaluated by Western blot and RT-PCR , respectively , while only one was positive for P35354 . The cytostatic and cytocidal activity was determined by sulforhodamine B assay and evaluated according to Monks ' model . Cytostatic activity was observed after a 24-h drug exposure and 50 % growth inhibition was reached at concentrations ranging from 165 to 250 micro M in all cell lines , whereas with aspirin the IC50 was never reached , even at the maximum concentration tested ( 500 micro M ) , and was independent of P23219 or P35354 status . Cytocidal activity was observed only at the highest concentrations and persisted for a long time after drug removal . Flow cytometric analysis showed that the NO-aspirin compound induced a persistent accumulation of cells in G2-M phase in all the cell lines after at least 48 h exposure . Specifically , the block pertained mainly to G2 phase , whereas mitotic index was not affected at all . Our results indicate that DB05822 has an in vitro cytostatic activity superior to that of its parental aspirin compound , which makes it a potentially important tumor preventive agent . Furthermore , the cytocidal effect observed at the highest concentrations and the induction of a specific block in G2 phase renders it a promising candidate for drug combination treatments . ICE/ P29466 inhibitors as novel anti-inflammatory drugs . In recent years , several strategies that selectively inhibit pro-inflammatory cytokines , have yielded effective protein-based therapies for inflammatory disorders , validating the therapeutic hypothesis that intervention in cytokine signalling can provide clinical benefit . However , these protein-based products must be administered by injection , a constraint associated with inconvenience , adverse effects and expense for patients , caregivers and insurers . Besides interfering with the effects of cytokines such as P01375 or IL-1beta that have already been produced , inhibition of pro-inflammatory cytokine production or signalling with low-molecular weight orally-active drugs would combine the convenience of conventional pharmaceuticals with the focused efficacy of the protein therapies . Reducing IL-1beta and Q14116 production by inhibition of IL-1beta converting enzyme ( ICE , caspase-1 ) is one promising strategy because of the key roles of these cytokines in many inflammatory diseases . DB04875 , the first orally available , potent and selective ICE inhibitor to enter clinical trials , is currently under investigation in rheumatoid arthritis . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . Permanent neonatal diabetes mellitus in China . BACKGROUND : Permanent neonatal diabetes mellitus ( PNDM ) is a rare disease , which is defined as the onset of diabetes before the age of 6 months with persistence through life . Infants with Q14654 or Q09428 genetic mutations may respond to oral sulfonylurea therapy . Currently , there are limited studies about the genetic analysis and long-term follow-up of PNDM . CASE PRESENTATION : We report four cases of PNDM . None of the infants or their parents had P01308 , Q14654 , or Q09428 genetic mutations . One infant underwent continuous subcutaneous insulin infusion ( CSII ) and the other infants underwent multiple injections of insulin ( MII ) . In these infants , PNDM persisted from 35 months to 60 months of follow-up . Three infants maintained fairly stable blood sugar levels , and one infant had poor sugar control . CONCLUSIONS : We suggest that all of the infants with PNDM should undergo genetic evaluation . For infants without Q14654 and Q09428 genetic mutations , oral sulfonylurea should not be considered as treatment . CSII is a useful method for overcoming the difficulties of diabetes , and it may also improve the quality of life of both infants and their parents . [ Conditions of the primary culture for rat hepatocytes and plasminogen activator release ] . The conditions of primary culture for rat hepatocytes was investigated on the releasing effect of P00747 Activator ( PA ) . The culture method using Collagen Coated Dish ( CCD-method ) which is currently available and the ordinary culture method using Plastic Culture Dish ( P61457 -method ) were employed for that purpose in a comparative way . The effect of the addition of some supplements , that is FN , DB06692 , P01133 were also investigated . The following results were obtained . The dissociated rat hepatocytes formed a monolayer with pavementlike morphology at 24-48 hours after seeding . No difference was observed in the morphology of hepatocytes during the culture period between the two methods , although CCD-method allowed 120 hours culture , whereas P61457 -method allowed 72 hours . The PA activity was demonstrated on the hepatocytes by either culture method according to the fibrinolysis autography . The cultured hepatocytes released PA into the medium continuously as long as the viability and morphology of the cells were maintained in good state . The PA activity reached the maximum after 96 hours culture in CCD-method , whereas it reached the maximum after 48 hours in P61457 -method . The addition of DB06692 to the culture medium was not necessary for PA release in CCD-method in contrast to P61457 -method . When P01133 was discontinued in the culture medium , the release of PA was reduced in association with the occurring of morphological disintegration of hepatocytes . Evaluation of [(11)C]rofecoxib as PET tracer for cyclooxygenase 2 overexpression in rat models of inflammation . BACKGROUND : Overexpression of cyclooxygenase type 2 ( P35354 ) is triggered by inflammatory stimuli , but it also plays a prominent role in the initiation and progression of various diseases . This study aims to investigate [(11)C]rofecoxib as a positron emission tomography ( PET ) tracer for P35354 expression . METHODS : [(11)C] DB00533 was prepared by methylation of its sulphinate precursor . Regional brain distribution and specific binding of [(11)C]rofecoxib in healthy rats was studied by ex vivo biodistribution and autoradiography . Regional brain distribution and PET imaging studies were also performed on rats with severe encephalitis , caused by nasal infection with herpes simplex virus ( HSV ) . Finally , ex vivo biodistribution and blocking studies were carried in rats with a sterile inflammation , induced by intramuscular turpentine injection . RESULTS : [(11)C]rofecoxib brain uptake in control animals corresponded with the known distribution of P35354 . Pretreatment with NS398 significantly reduced tracer uptake in the cingulate/frontopolar cortex , whereas the reduction in hippocampus approached significance . Ex vivo autoradiography also revealed preferential tracer uptake in hippocampus and cortical areas that could be blocked by NS398 . In HSV-infected animals , [(11)C]rofecoxib uptake was moderately increased in all brain regions , but it could not be blocked with indomethacin . Yet , some PET images revealed increased tracer uptake in brain areas with microglia activation . In turpentine-injected animals , [(11)C]rofecoxib uptake in inflamed muscle was not higher than in control muscle and could not be blocked with NS398 . Indomethacin caused a slight reduction in muscle uptake . CONCLUSIONS : Despite the apparent correlation between [(11)C]rofecoxib uptake and P35354 distribution in healthy rats , [(11)C]rofecoxib could not unambiguously detect P35354 overexpression in two rat models of inflammation . P14735 binds to the nonglycosylated precursor of varicella-zoster virus gE protein found in the endoplasmic reticulum . P01308 degradation enzyme ( P14735 ) is a 110-kDa zinc metalloprotease found in the cytosol of all cells . P14735 degrades insulin and a variety of small proteins including amyloid-beta . Recently , P14735 has been proposed as the receptor for varicella-zoster virus ( VZV ) attachment . During our reassessment , some of the original studies were repeated and expanded in scope . We first confirmed that P14735 antibody reduced VZV spread . For additional controls , we repeated the same experiments with herpes simplex virus ( HSV ) -infected cells as well as uninfected cells . There was a visible reduction in HSV spread but less than seen in the VZV system . Of greater importance , P14735 antibody also inhibited the growth of uninfected cells . Second , we repeated the coprecipitation assays . We confirmed that antibodies to VZV gE ( open reading frame 68 ) coprecipitated P14735 and that anti- P14735 antibody coprecipitated gE . However , the detected gE protein was not the mature 98-kDa form ; rather , it was a precursor 73-kDa gE form found in the endoplasmic reticulum . Additional control experiments included VZV-infected cell cultures treated with tunicamycin to block gE glycosylation in the endoplasmic reticulum ; again , the anti- P14735 antibody coprecipitated a 73-kDa gE product . Finally , Orbitrap mass spectrometry analysis of a chromatographically purified gE sample revealed four cellular proteins associated with the unfolded protein response : P11021 ( P11021 ) , P11142 , P10809 , and P62937 ( peptidyl-propyl cis-trans isomerase ) . We conclude that P14735 protease binds to the 73-kDa gE precursor and that this event occurs in the cytosol but not as a receptor/ligand interaction . K( DB00171 ) channel openers in the trigeminovascular system . BACKGROUND : The DB00171 -sensitive K(+) ( K( DB00171 ) ) channel openers levcromakalim and pinacidil are vasodilators that induce headache in healthy people . The neuropeptide calcitonin gene-related peptide ( P80511 ) induces headache in healthy people and migraine in migraineurs , potentially through a mechanism that involves opening of vascular or neuronal K( DB00171 ) channels and mast cell degranulation . Using rat as a model , we studied the molecular presence of K( DB00171 ) channels in the trigeminovascular system . Furthermore , we examined whether K( DB00171 ) channel openers stimulate the in vitro release of P80511 and whether they degranulate dural mast cells . METHODS : mRNA and protein expression of K( DB00171 ) channel subunits were studied in the trigeminal ganglion ( TG ) and trigeminal nucleus caudalis ( P24821 ) by qPCR and western blotting . In vitro P80511 release was studied after application of levcromakalim ( 1 µM ) and diazoxide ( 10 µM ) to freshly isolated rat dura mater , TG and P24821 . Rat dural mast cells were challenged in situ with levcromakalim ( 10(-5) M ) to study its potential degranulation effect . RESULTS : mRNA and protein of K( DB00171 ) channel subunits Kir6.1 , Kir6.2 , Q09428 and SUR2B were identified in the TG and P24821 . K( DB00171 ) channel openers did not release or inhibit capsaicin-induced P80511 release from dura mater , TG or P24821 . They did also not induce dural mast cell degranulation . CONCLUSIONS : K( DB00171 ) channel openers do not interact with P80511 release or mast cell degranulation . Activation of these channels in the CNS is antinociceptive and therefore can not explain the headache induced by K( DB00171 ) channel openers . Thus , they are likely to induce headache by interaction with extracerebral K( DB00171 ) channels , probably the SUR2B isoforms . P62937 as a target of DB00515 chemosensitizers . Platinum-based chemotherapeutics are the mainstay of treatment of a range of tumors achieving high response rates but limited in the course of disease by appearance of drug resistance . Tumor cells respond with reduced uptake and increased intracellular inactivation of the drugs , as well as increased DNA repair and general resistance to chemotherapyinduced cell death . DB00515 is known to induce expression of cyclophilins , a group of proteins that have peptidyl-prolyl cis-trans isomerase ( PPIase ) and molecular chaperone activities , as stress response . P62937 ( CypA ) and other members of this family are inhibited by cyclosporin A ( DB00091 ) which sensitized diverse drug-resistant tumor cell lines in vitro to cisplatin . This effect of DB00091 was attributed to metabolic changes , inhibition of DNA repair , enhancement of apoptosis , altered intracellular signal transduction or increased production of reactive oxygen species ( ROS ) , although no definitive explanation was provided so far . Several clinical trials employing cisplatin/carboplatin in combination with DB00091 yielded unsatisfactory results . Since viral replication was found to be dependent on cyclophilins of the host cells , effective new inhibitors , different from DB00091 or with low or absent immunosuppressive activity , are in development or clinical trials . Sanglifehrins are more potent than DB00091 and proved to increase toxicity of cisplatin against hepatocellular cancer cells in vitro . These novel cyclophilin inhibitors may offer new opportunities to achieve reversal of resistance to platinumbased drugs in refractory patients . Responsive cancer patients may be enriched in clinical trials by an identification of the downstream targets of Cyps responsible for chemoresistance . Modulation of arsenic-induced epidermal growth factor receptor pathway signalling by resveratrol . Arsenic ( As ) is both a human carcinogen and an effective anticancer drug . These aspects of arsenic toxicity develop as a consequence of arsenic-induced oxidative stress and modifications to signal pathway activity which alter gene expression . DB02709 ( RVL ) a food antioxidant found in grapes and other fruits , exhibits anti-carcinogenic properties by reducing oxidative stress and restoring signal pathway control . This study investigated the impact of RVL on arsenite [ As(III) ] -induced cell signalling in HaCaT keratinocytes by assaying phosphorylation status of epidermal growth factor receptor ( P00533 ) signalling intermediates and measuring changes in expression of Phase II and DNA repair biomarkers . As(III) exposure produced dose-dependent toxicity which was associated with increased activation of P00533 pathway intermediates , cSrc , Rac1 and extracellular signal-regulated kinases 1 and 2 ( P27361 /2 ) . Arsenic-mediated P27361 /2 activation negatively regulated P06746 expression and up regulated heme-oxygenase-1 at toxic concentrations . RVL treatment modulated As(III)-mediated P27361 /2 activation by shifting the balance of cSrc regulatory domain phosphorylation . These effects significantly altered the response of the P00533 pathway to growth factor-induced stimulation . Our research provides evidence that treatment with pharmacologically relevant doses of RVL influences cellular responses to As(III) , largely due to RVL-mediated changes to Src and P27361 /2 activation . Pancreatic hypoplasia presenting with neonatal diabetes mellitus in association with congenital heart defect and developmental delay . Congenital pancreatic hypoplasia is a rare cause of neonatal diabetes . We report on a series of three patients with pancreatic agenesis and congenital heart defects . All had abdominal scan evidence of pancreatic agenesis . In addition , Patient 1 had a ventricular septal defect , patent ductus arteriosus and pulmonary artery stenosis ; Patient 2 had a truncus arteriosus and Patient 3 had tetralogy of Fallot . Two of the three patients have developmental delay . All three patients were isolated cases within the family . Investigations included sequencing of GCK , Q09428 , P52945 , Q13562 , Q7RTS3 , P35680 , P01308 , P61371 , Q9Y4Z2 , Q03014 , Q9NQR9 , Q9NQB0 , Q06945 , Q9BZS1 ( Patients 1 and 2 ) , P43694 and Q14654 genes ( all three patients ) , but no mutations were found . Genetic investigation to exclude paternal P06132 6 , methylation aberrations and duplications of 6q24 was also negative in all three . 22q11 deletion was excluded in all three patients . Array CGH in Patient ( 1 ) showed a approximately 250 kb , paternally inherited duplication of chromosome 12q [ arr cgh 12q24.33 ( B35:CHR12:131808577-132057649++ ) pat ] , not found in the other two patients . Permanent neonatal diabetes mellitus due to pancreatic hypoplasia with congenital heart defects has been reported before and may represent a distinct condition . We discuss this rare association and review previously reported literature . Silencing of ALA dehydratase affects ALA-photodynamic therapy efficacy in K562 erythroleukemic cells . Synthesis of protoporphyrin IX ( PpIX ) by malignant cells is essential for the success of ALA-based photodynamic therapy ( PDT ) . Two key enzymes that were described as affecting PpIX accumulation during ALA treatment are porphobilinogen deaminase ( P08397 ) and ferrochelatase . Here , we show that down regulation of ALA dehydratase ( P13716 ) expression and activity by specific shRNA induced a marked decrease in PpIX synthesis in K562 erythroleukemic cells . Photo-inactivation efficacy following DB00855 was directly correlated with P13716 -silencing and cellular levels of PpIX . MTT metabolism following DB00855 was shown to be 60 % higher in P13716 -silenced cells in comparison to control cells , indicating that mitochondria were protected in the silenced cells . Morphological analysis by scanning electron microscopy ( SEM ) of cells treated by DB00855 showed no morphological changes in P13716 -silenced cells , in contrast to controls exhibiting cell deformations and lysis . Membrane integrity following DB00855 was kept intact and undamaged in P13716 -silenced cells as examined by P08758 -FITC/PI staining and LDH-L leakage . We conclude that P13716 , although it is present in the cell at abundant levels , has a major and limiting role in regulating PpIX synthesis and DB00855 outcome . Q07869 controls hepatic heme biosynthesis through P13196 . Heme is an essential prosthetic group of proteins involved in oxygen transport , energy metabolism and nitric oxide production . P13196 ( 5-aminolevulinate synthase ) is the rate-limiting enzyme in heme synthesis in the liver and is highly regulated to adapt to the metabolic demand of the hepatocyte . In the present study , we describe human hepatic P13196 as a new direct target for the nuclear receptor peroxisome proliferator-activated receptor alpha ( PPARalpha ) . In primary human hepatocytes and in HepG2 cells , PPARalpha agonists induced an increase in P13196 mRNA levels , which was abolished by PPARalpha silencing . These effects are mediated by two functional Q07869 binding sites at positions -9 and -2.3 kb relative to the P13196 transcription start site . PPARalpha ligand treatment also up-regulated the mRNA levels of the genes P13716 ( 5-aminolevulinate dehydratase ) , P10746 ( uroporphyrinogen III synthase ) , P06132 ( uroporphyrinogen decarboxylase ) , P36551 ( coproporphyrinogen oxidase ) and PPOX ( protoporphyrinogen oxidase ) encoding for enzymes controlling further steps in heme biosynthesis . In HepG2 cells treated with PPARalpha agonists and in mouse liver upon fasting , the association of PPARalpha , its partner retinoid X receptor , PPARgamma co-activator 1alpha and activated RNA polymerase II with the transcription start site region of all six genes was increased , leading to higher levels of the metabolite heme . In conclusion , these data strongly support a role of PPARalpha in the regulation of human P13196 and of five additional genes of the pathway , consequently leading to increased heme synthesis . Anti-clastogenic effect of beta-glucan extracted from barley towards chemically induced DNA damage in rodent cells . beta-Glucan ( BG ) was tested in vitro to determine its potential clastogenic and/or anti-clastogenic activity , and attempts were made to elucidate its possible mechanism of action by using combinations with an inhibitor of DNA polymerase . The study was carried out on cells deficient ( CHO-k1 ) and cells proficient ( HTC ) in phases I and II enzymes , and the DNA damage was assessed by the chromosomal aberration assay . BG did not show a clastogenic effect , but was anti-clastogenic in both cell lines used , and at all concentrations tested ( 2.5 , 5 and 10 microg/mL ) in combination with damage inducing agents ( methylmethane sulfonate in cell line CHO-k1 , and methylmethane sulfonate or 2-aminoanthracene in cell line HTC ) . BG also showed a protective effect in the presence of a P06746 inhibitor ( cytosine arabinoside-3-phosphate , DB00987 ) , demonstrating that BG does not act through an anti-mutagenic mechanism of action involving P06746 .

MH_train_1332

MH_train_1332

MH_train_1332

interacts_with DB08907?

multiple_choice

Isoenzyme-specific cyclooxygenase inhibitors : a whole cell assay system using the human erythroleukemic cell line HEL and the human monocytic cell line Mono Mac 6 . NSAIDs inhibit the conversion of arachidonic acid into DB03866 and Prostaglandin H2 which is catalyzed by the enzyme cyclooxygenase ( P36551 ) . Two genetically distinct isoforms have been discovered , P23219 and P35354 . While P23219 is thought to account for homeostatic amounts of eicosanoids , P35354 is induced during inflammation leading to pathologic amounts of eicosanoids . Since NSAIDs inhibit both P36551 isoforms , antiinflammatory drug research has refocused to discovering P35354 inhibitors that do not inhibit P23219 . For this purpose , we have developed a whole cell assay system using the human erythroleukemic cell line HEL as a source for P23219 and the human monocytic cell line Mono Mac 6 as a source for P35354 . Mono Mac 6 cells express high amounts of P35354 upon stimulation with lipopolysaccharide ( LPS ) in the absence of any detectable P23219 protein . On the other hand , we find HEL cells to naturally express P23219 protein , but not P35354 . Testing of a panel of NSAIDs as well as some P35354 specific inhibitors showed that this assay system is suitable for identifying compounds that selectively inhibit either P23219 or P35354 . This test system offers the advantage of assessing P23219 and P35354 inhibitors within the human species , within a similar test set-up , and circumvents the need for tedious purification of either platelets or peripheral blood monocytes . Estrogen response element-dependent regulation of transcriptional activation of estrogen receptors alpha and beta by coactivators and corepressors . One mechanism by which ligand-activated estrogen receptors alpha and beta ( ERalpha and ERbeta ) stimulate gene transcription is through direct ER interaction with specific DNA sequences , estrogen response elements ( EREs ) . ERE-bound ER recruits coactivators that stimulate gene transcription . Binding of ER to natural and synthetic EREs with different nucleotide sequences alters ER binding affinity , conformation , and transcriptional activity , indicating that the ERE sequence is an allosteric effector of ER action . Here we tested the hypothesis that alterations in ER conformation induced by binding to different ERE sequences modulates ER interaction with coactivators and corepressors . CHO- P04264 cells transfected with ERalpha or ERbeta show ERE sequence-dependent differences in the functional interaction of ERalpha and ERbeta with coactivators steroid receptor coactivator 1 ( Q15788 ) , P12931 -2 ( glucocorticoid receptor interacting protein 1 ( GRIP1 ) ) , Q9Y6Q9 amplified in breast cancer 1 ( Q9Y6Q9 ) and Q9Y6Q9 , cyclic AMP binding protein ( CBP ) , and steroid receptor RNA activator ( SRA ) , corepressors nuclear receptor co-repressor ( NCoR ) and silencing mediator for retinoid and thyroid hormone receptors ( Q9Y618 ) , and secondary coactivators coactivator associated arginine methyltransferase 1 ( Q86X55 ) and protein arginine methyltransferase 1 ( Q99873 ) . We note both ligand-independent as well estradiol- and 4-hydroxytamoxifen-dependent differences in ER-coregulator activity . In vitro ER-ERE binding assays using receptor interaction domains of these coregulators failed to recapitulate the cell-based results , substantiating the importance of the full-length proteins in regulating ER activity . These data demonstrated that the ERE sequence impacts estradiol-and 4-hydroxytamoxifen-occupied ERalpha and ERbeta interaction with coregulators as measured by transcriptional activity in mammalian cells . Effect of canagliflozin on renal threshold for glucose , glycemia , and body weight in normal and diabetic animal models . BACKGROUND : DB08907 is a sodium glucose co-transporter ( SGLT ) 2 inhibitor in clinical development for the treatment of type 2 diabetes mellitus ( T2DM ) . METHODS : (14)C-alpha-methylglucoside uptake in Chinese hamster ovary-K cells expressing human , rat , or mouse SGLT2 or P13866 ; (3)H-2-deoxy-d-glucose uptake in Q9BTT4 myoblasts ; and 2-electrode voltage clamp recording of oocytes expressing human SGLT3 were analyzed . Graded glucose infusions were performed to determine rate of urinary glucose excretion ( UGE ) at different blood glucose ( BG ) concentrations and the renal threshold for glucose excretion ( RT(G) ) in vehicle or canagliflozin-treated Zucker diabetic fatty ( ZDF ) rats . This study aimed to characterize the pharmacodynamic effects of canagliflozin in vitro and in preclinical models of T2DM and obesity . RESULTS : Treatment with canagliflozin 1 mg/kg lowered RT(G) from 415±12 mg/dl to 94±10 mg/dl in ZDF rats while maintaining a threshold relationship between BG and UGE with virtually no UGE observed when BG was below RT(G) . DB08907 dose-dependently decreased BG concentrations in db/db mice treated acutely . In ZDF rats treated for 4 weeks , canagliflozin decreased glycated hemoglobin ( HbA1c ) and improved measures of insulin secretion . In obese animal models , canagliflozin increased UGE and decreased BG , body weight gain , epididymal fat , liver weight , and the respiratory exchange ratio . CONCLUSIONS : DB08907 lowered RT(G) and increased UGE , improved glycemic control and beta-cell function in rodent models of T2DM , and reduced body weight gain in rodent models of obesity . Microarray analysis reveals characteristic changes of host cell gene expression in response to attenuated modified vaccinia virus Ankara infection of human HeLa cells . The potential use of the modified vaccinia virus Ankara ( MVA ) strain as a live recombinant vector to deliver antigens and elicit protective immune responses against infectious diseases demands a comprehensive understanding of the effect of MVA infection on human host gene expression . We used microarrays containing more than 15,000 human cDNAs to identify gene expression changes in human HeLa cell cultures at 2 , 6 , and 16 h postinfection . Clustering of the 410 differentially regulated genes identified 11 discrete gene clusters with altered expression patterns after MVA infection . Clusters 1 and 2 ( accounting for 16.59 % [ 68 of 410 ] of the genes ) contained 68 transcripts showing a robust induction pattern that was maintained during the course of infection . Changes in cellular gene transcription detected by microarrays after MVA infection were confirmed for selected genes by Northern blot analysis and by real-time reverse transcription-PCR . Upregulated transcripts in clusters 1 and 2 included 20 genes implicated in immune responses , including interleukin 1A ( IL-1A ) , P05231 , P13232 , P10145 , and P40933 genes . MVA infection also stimulated the expression of NF-kappaB and components of the NF-kappaB signal transduction pathway , including p50 and TRAF-interacting protein . A marked increase in the expression of histone family members was also induced during MVA infection . Expression of the Wiskott-Aldrich syndrome family members P42768 , Q92558 , and the small GTP-binding protein P31749 -1 , which are involved in actin cytoskeleton reorganization , was enhanced after MVA infection . This study demonstrates that MVA infection triggered the induction of groups of genes , some of which may be involved in host resistance and immune modulation during virus infection . DB02342 in the human corpus luteum throughout the luteal phase and its influence on lutein cell steroidogenesis and angiogenic activity . OBJECTIVE : To quantitate 2-methoxyestradiol ( 2-ME ) in human corpus luteum ( CL ) of different ages and to determine the expression of cytochrome-P450-1A1 ( P04798 ) and catechol-O-methyl transferase ( P21964 ) in CL and the action of 2-ME on P , vascular endothelial growth factor ( P15692 ) secretion , and luteal angiogenesis . DESIGN : Experimental study . SETTING : University division of reproductive endocrinology . PATIENT(S) : Twenty-four women of reproductive age . INTERVENTION(S) : CL was collected from 15 women during the minilaparotomy for tubal sterilization . Granulosa lutein cells were harvested 36 hours after hCG administration in patients undergoing IVF . MAIN OUTCOMES MEASURE(S) : Levels of 2-ME were determined by high-performance liquid chromatography in CL . P04798 and P21964 were assessed by immunohistochemistry and Western blot . P and P15692 were measured by radioimmunoassay and ELISA . The angiogenic potential was analyzed using EA.hy926 cells . RESULT(S) : Plasma levels of E₂ decreased in the late luteal phase in association with an increase in luteal tissue of 2-ME concentrations . Concomitantly , there was a significant reduction of angiogenic activity in late CL . There was no significant variation in P04798 and P21964 expression in all CL . In physiological doses , 2-ME inhibited basal P15692 by granulosa lutein cells and diminished the angiogenic activity in conditioned media but did not prevent P and P15692 production stimulated by hCG . CONCLUSION(S) : These data suggest the participation of 2-ME in physiological luteolysis by reducing angiogenesis . However , 2-ME did not prevent in vitro hCG stimulation of P biosynthesis , providing a mechanism for CL rescue in the cycle of conception . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . Identification of Reverb(alpha) as a novel ROR(alpha) target gene . The nuclear receptor superfamily comprises a large number of ligand-activated transcription factors that are involved in numerous biological processes such as cell proliferation , differentiation , and homeostasis . ROR(alpha) ( P35398 ) and Reverb(alpha) ( P20393 ) are two members of this family whose biological functions are largely unknown . In addition , no ligand has been yet identified for these two receptors ; therefore , they are referred as orphan receptors . Here , we show that ROR(alpha) and Reverb(alpha) are expressed with a similar tissue distribution and are both induced during the differentiation of rat Q9BTT4 myoblastic cells . Ectopic expression of ROR(alpha)1 in Q9BTT4 cells significantly induces Reverb(alpha) expression as demonstrated by Northern blot analysis . Using reverse transcription-PCR to analyze Reverb(alpha) gene expression from staggerer mice , we found that there was a significant reduction of Reverb(alpha) mRNA in the skeletal muscle comparing it with the wild-type mice , which suggests that ROR(alpha) is involved in the regulation of Reverb(alpha) gene expression . Transient transfection assays using the Reverb(alpha) promoter demonstrate that ROR(alpha) regulates the Reverb(alpha) gene at the transcriptional level . Furthermore , mutagenesis experiments indicate that ROR(alpha) regulates Reverb(alpha) transcription via a monomeric ROR response element located in the Reverb(alpha) gene promoter . Electrophoretic mobility shift assays show that ROR(alpha) binds strongly to this site in a specific-manner . Finally , overexpression of Q9Y3R0 / Q06418 -2 , but not Q15788 , potentiates ROR(alpha)-stimulated Reverb(alpha) promoter activity in transient transfection experiments . Together , our results identify Reverb(alpha) as a novel target gene for ROR(alpha) . Neuronal specific increase of phosphatidylserine by docosahexaenoic acid . DB00144 ( PS ) , the major acidic phospholipid class in eukaryotic biomembranes , plays an important role in various signaling pathways . We have previously demonstrated that docosahexaenoic acid ( DB01708 , 22:6n-3 ) positively modulates PS biosynthesis and accumulation in neuronal cells , promoting survival . In this paper , we demonstrate that the increase of PS levels upon DB01708 enrichment is not a universal mechanism , but specific to neuronal cells . When cells were enriched with 20 muM DB01708 , 18:0 , 22:6-PS increased in both neuronal ( Neuro 2A ) and non-neuronal cells ( Chinese hamster ovary P04264 cells , NIH-3T3 , and human embryonic kidney cells ) . However , the increase of the total PS level was observed only in Neuro 2A cells because of the fact that other PS species , such as 18:0 , 18:1-PS and 18:1 , 18:1-PS decreased significantly in non-neuronal cells , compensating for the increase of 18:0 , 22:6-PS . DB01708 enrichment did not affect the messenger RNA levels of PS synthase 1 ( PSS1 ) and Q9BVG9 . Over-expression of genes encoding PSS1 or Q9BVG9 altered neither the PS level nor the effect of DB01708 on PS increase in both neuronal and non-neuronal cells . From these results , it is concluded that the PS increase by DB01708 , specifically observed in neuronal cells , may represent a unique mechanism for expanding the PS pool so far known in mammalian cells . Predicting the effect of naltrexone and acamprosate in alcohol-dependent patients using genetic indicators . DB00659 and naltrexone are effective medications in the treatment of alcoholism . However , effect sizes are modest . Pharmacogenomics may improve patient-treatment-matching and effect sizes . It is hypothesized that naltrexone exerts its effect through genetic characteristics associated with the dopaminergic/opioidergic positive reinforcement system , whereas acamprosate works through the glutamatergic/GABAergic negative reinforcement system . DB00898 -dependent subjects were randomly assigned to either acamprosate or naltrexone . Subjects participated in a cue-exposure experiment at the day before and at the last day of medication . Reductions in cue-induced craving and physiological cue reactivity were measured . Differential effects of naltrexone and acamprosate on these outcomes were tested for different polymorphisms of the opioid , dopamine , glutamate and GABA-receptors . Significant matching effects were found for polymorphisms at the P14416 , Q16445 and P47870 gene . In addition , a trend was found for the P35372 polymorphism . This provides evidence for the matching potential of genotypes . It is expected that more effective treatments can be offered when genetic information is used in patient-treatment-matching . Inactivation of caspase-1 in rodent brain : a novel anticonvulsive strategy . PURPOSE : Cytokines and related inflammatory mediators are rapidly synthesized in the brain during seizures . We previously found that intracerebral administration of interleukin-1 ( IL-1 ) -beta has proconvulsant effects , whereas its endogenous receptor antagonist ( IL-1Ra ) mediates potent anticonvulsant actions in various models of limbic seizures . In this study , we investigated whether seizures can be effectively inhibited by blocking the brain production of IL-1beta , by using selective inhibitors of interleukin-converting enzyme ( ICE/caspase-1 ) or through caspase-1 gene deletion . METHODS : P29466 was selectively blocked by using pralnacasan or DB05507 . IL-1beta release was induced in mouse organotypic hippocampal slice cultures by proinflammatory stimuli [ lipopolysaccharide ( LPS ) + adenosine triphosphate ( DB00171 ) ] and measured with enzyme-linked immunosorbent assay ( ELISA ) . IL-1beta production during seizures was measured in the rat hippocampus by Western blot . Seizures were induced in freely moving mice and rats by intrahippocampal injection of kainic acid and recorded by EEG analysis . RESULTS : P29466 inhibition reduced the release of IL-1beta in organotypic slices exposed to LPS+ DB00171 . Administration of pralnacasan ( intracerebroventricular , 50 microg ) or DB05507 ( intraperitoneal , 25-200 mg/kg ) to rats blocked seizure-induced production of IL-1beta in the hippocampus , and resulted in a twofold delay in seizure onset and 50 % reduction in seizure duration . Mice with caspase-1 gene deletion showed a 70 % reduction in seizures and an approximate fourfold delay in their onset . CONCLUSIONS : Inhibition of caspase-1 represents an effective and novel anticonvulsive strategy , which acts by selectively reducing the brain availability of IL-1beta . Identification and characterization of nucleolin as a COUP-TFII coactivator of retinoic acid receptor β transcription in breast cancer cells . INTRODUCTION : The orphan nuclear receptor COUP-TFII plays an undefined role in breast cancer . Previously we reported lower COUP-TFII expression in tamoxifen/endocrine-resistant versus sensitive breast cancer cell lines . The identification of COUP-TFII-interacting proteins will help to elucidate its mechanism of action as a transcriptional regulator in breast cancer . RESULTS : FLAG-affinity purification and multidimensional protein identification technology ( MudPIT ) identified nucleolin among the proteins interacting with COUP-TFII in MCF-7 tamoxifen-sensitive breast cancer cells . Interaction of COUP-TFII and nucleolin was confirmed by coimmunoprecipitation of endogenous proteins in MCF-7 and T47D breast cancer cells . In vitro studies revealed that COUP-TFII interacts with the C-terminal arginine-glycine repeat ( RGG ) domain of nucleolin . Functional interaction between COUP-TFII and nucleolin was indicated by studies showing that siRNA knockdown of nucleolin and an oligonucleotide aptamer that targets nucleolin , DB04998 , inhibited endogenous COUP-TFII-stimulated RARB2 expression in MCF-7 and T47D cells . Chromatin immunoprecipitation revealed COUP-TFII occupancy of the RARB2 promoter was increased by all-trans retinoic acid ( atRA ) . RARβ2 regulated gene RRIG1 was increased by atRA and COUP-TFII transfection and inhibited by siCOUP-TFII . Immunohistochemical staining of breast tumor microarrays showed nuclear COUP-TFII and nucleolin staining was correlated in invasive ductal carcinomas . COUP-TFII staining correlated with ERα , Q15788 , Q9Y6Q9 , Pea3 , P08253 , and phospho-Src and was reduced with increased tumor grade . CONCLUSIONS : Our data indicate that nucleolin plays a coregulatory role in transcriptional regulation of the tumor suppressor RARB2 by COUP-TFII . DB08890 - a secretagogue and antihyperalgesic agent - what next ? Ongoing clinical trials suggest that linaclotide , a first-in-class , 14-amino acid peptide guanylate cyclase-C ( P25092 ) receptor agonist and intestinal secretagogue is an effective treatment for chronic constipation . A study in this issue of the Journal suggests that linaclotide also has antihyperalgesic effects in three common rat models of inflammation- and stress-induced hypersensitivity ( i.e. , acute trinitrobenzene sulfonic acid colitis , water avoidance stress [ P42768 ] , and restraint-induced stress ) but not in naïve animals . In mice , linaclotide at least partly reduces hyperalgesia via P25092 receptors . Dose-effect relationships of linaclotide were complicated and non-linear . This viewpoint discusses human clinical trials with linaclotide and the results of this study . Potential mechanisms and clinical significance of these findings are explored . Collectively , these data suggest that P25092 receptors exert other , as yet poorly understood , effects on gastrointestinal sensitivity in conditions associated with inflammation and/or stress-induced increased intestinal permeability . However , the data need to be confirmed in humans and in long-term animal models . Further studies are also necessary to elucidate the mechanisms as these effects can not be explained by linaclotide 's known effects on epithelial P25092 receptors . Inhibition of matrix metalloproteinase-2 by halofuginone is mediated by the Egr1 transcription factor . DB04866 , a low-molecular-weight quinazolinone alkaloid that inhibits collagen α1(I) , has been shown to suppress cancer growth , metastasis , and angiogenesis . These activities were attributed in part to the inhibition of matrix metalloproteinase-2 ( P08253 ) . The present study was carried out to explore the molecular mechanism underlying this effect . We found a marked ( 50 % ) inhibition in P08253 gelatinolytic activity in human breast cancer MDA-MB-435 cells pretreated with as little as 50 ng/ml of halofuginone , a concentration that markedly inhibited their invasive and proliferative capacities . We further show that both early growth response 1 ( Egr-1 ) and Nab-2 ( corepressor of Egr1 activation ) are upregulated by halofuginone in a dose-dependent and time-dependent ( up to 5 h ) manner . Using P08253 reporter gene and chromatin immunoprecipitation analyses , we found that Egr-1 binds to the P08253 promoter and inhibits its activity . Altogether , our results identify the downstream elements ( Egr-1 , Nab-2 , and P08253 ) by which halofuginone exerts its antitumoral effect , thereby advancing its potential therapeutic application as an anticancer drug . Zebrafish express the common parathyroid hormone/parathyroid hormone-related peptide receptor ( Q03431 ) and a novel receptor ( PTH3R ) that is preferentially activated by mammalian and fugufish parathyroid hormone-related peptide . To further explore the evolution of receptors for parathyroid hormone ( PTH ) and PTH-related peptide ( P12272 ) , we searched for zebrafish ( z ) homologs of the PTH/ P12272 receptor ( Q03431 ) . In mammalian genes encoding this receptor , exons M6/7 and M7 are highly conserved and separated by 81-84 intronic nucleotides . Genomic polymerase chain reaction using degenerate primers based on these exons led to two distinct DNA fragments comprising portions of genes encoding the zPTH1R and the novel zPTH3R . Sequence comparison of both full-length teleost receptors revealed 69 % similarity ( 61 % identity ) , but less homology with zPTH2R . When compared with hPTH1R , zPTH1R showed 76 % and zPTH3R 67 % amino acid sequence similarity ; similarity with hPTH2R was only 59 % for both teleost receptors . When expressed in COS-7 cells , zPTH1R bound [ DB00135 (34)] DB05829 -(1-34)-amide ( DB05829 ) , [ DB00135 (36)]hPTHrP-(1-36)-amide ( hPTHrP ) , and [ Ala(29), DB00142 (30) , Ala(34), DB00142 (35) , DB00135 (36) ] fugufish P12272 -(1-36)-amide ( fuguPTHrP ) with a high apparent affinity ( IC(50) : 1.2-3.5 nM ) , and was efficiently activated by all three peptides ( EC(50) : 1.1-1.7 nM ) . In contrast , zPTH3R showed higher affinity for fuguPTHrP and hPTHrP ( IC(50) : 2.1-11.1 nM ) than for DB05829 ( IC(50) : 118.2-127.0 nM ) ; DB02527 accumulation was more efficiently stimulated by fugufish and human P12272 ( EC(50) : 0.47 +/- 0.27 and 0.45 +/- 0.16 , respectively ) than by DB05829 ( EC(50) : 9.95 +/- 1.5 nM ) . Agonist-stimulated total inositol phosphate accumulation was observed with zPTH1R , but not zPTH3R . A structural and in vitro characterization of asoprisnil : a selective progesterone receptor modulator . Selective progesterone receptor modulators ( SPRMs ) have been suggested as therapeutic agents for treatment of gynecological disorders . One such SPRM , asoprisnil , was recently in clinical trials for treatment of uterine fibroids and endometriosis . We present the crystal structures of progesterone receptor ( PR ) ligand binding domain complexed with asoprisnil and the corepressors nuclear receptor corepressor ( NCoR ) and Q9Y618 . This is the first report of steroid nuclear receptor crystal structures with ligand and corepressors . These structures show PR in a different conformation than PR complexed with progesterone ( P4 ) . We profiled asoprisnil in PR-dependent assays to understand further the PR-mediated mechanism of action . We confirmed previous findings that asoprisnil demonstrated antagonism , but not agonism , in a PR-B transfection assay and the T47D breast cancer cell alkaline phosphatase activity assay . Asoprisnil , but not DB00834 , weakly recruited the coactivators Q15788 and Q9Y6Q9 . However , asoprisnil strongly recruited the corepressor NCoR in a manner similar to DB00834 . Unlike DB00834 , NCoR binding to asoprisnil-bound PR could be displaced with equal affinity by NCoR or Q15596 peptides . We further showed that it weakly activated T47D cell gene expression of Sgk-1 and O60437 and antagonized P4-induced expression of both genes . In rat leiomyoma ELT3 cells , asoprisnil demonstrated partial P4-like inhibition of cyclooxygenase ( P36551 ) enzymatic activity and P35354 gene expression . In the rat uterotrophic assay , asoprisnil demonstrated no P4-like ability to oppose estrogen . Our data suggest that asoprisnil differentially recruits coactivators and corepressors compared to DB00834 or P4 , and this specific cofactor interaction profile is apparently insufficient to oppose estrogenic activity in rat uterus . The cyclooxygenase inhibitors indomethacin and DB00533 reduce regional cerebral blood flow evoked by somatosensory stimulation in rats . The present study was designed to investigate whether administration of indomethacin ( IMC ) , a non-selective cyclooxygenase ( P23219 and P35354 ) inhibitor , and DB00533 , a highly selective P35354 inhibitor , affect the regulation of regional cerebral blood flow response evoked by somatosensory activation ( evoked rCBF ) . IMC and DB00533 were applied intravenously ( 6.25 and 3 mg/kg/hr , respectively ) . Somatosensory activation was induced by electrical hind paw stimuli of 0.2 , 1 , and 5 Hz ( 5-sec duration , 1.5 mA ) . The evoked rCBF was measured in alpha-chloralose anesthetized rats using laser-Doppler flowmetry . Before and after drug application , the evoked rCBF showed a frequency-dependent increase in the range of 0.2-5 Hz stimulation . IMC reduced significantly ( about 50 % -60 % ) evoked rCBF in response to all frequencies of hind paw stimulation ( P < 0.05 ) . DB00533 reduced significantly ( about 50 % ) evoked rCBF in response to 1 and 5 Hz stimulation ( P < 0.05 ) , but did not affect evoked rCBF at 0.2 Hz . After IMC or DB00533 application , the normalized evoked rCBF curves peaked earlier as compared with that before their application ( P < 0.05 ) , although the rise time of 0.5 sec was nearly constant regardless of the stimulus frequency . The termination time of evoked rCBF curves was changed significantly after IMC application at 0.2 Hz stimulation ( P < 0.05 ) , but was not affected after DB00533 application . Neither P36551 inhibitor significantly affected the baseline level of Q03701 . The results suggest a participation of P36551 products in the regulation of evoked rCBF in response to somatosensory stimulation in the brain . Small molecule-driven mitophagy-mediated Q96P20 inflammasome inhibition is responsible for the prevention of colitis-associated cancer . Nonresolving inflammation in the intestine predisposes individuals to the development of colitis-associated cancer ( CAC ) . Inflammasomes are thought to mediate intestinal homeostasis , and their dysregulation contributes to inflammatory bowel diseases and CAC . However , few agents have been reported to reduce CAC by targeting inflammasomes . Here we show that the small molecule andrographolide ( Andro ) protects mice against azoxymethane/dextran sulfate sodium-induced colon carcinogenesis through inhibiting the Q96P20 inflammasome . Administration of Andro significantly attenuated colitis progression and tumor burden . Andro also inhibited Q96P20 inflammasome activation in macrophages both in vivo and in vitro , as indicated by reduced expression of cleaved P29466 , disruption of Q96P20 - Q9ULZ3 - P29466 complex assembly , and lower P01584 secretion . Importantly , Andro was found to trigger mitophagy in macrophages , leading to a reversed mitochondrial membrane potential collapse , which in turn inactivated the Q96P20 inflammasome . Moreover , downregulation of the P42336 - P31749 - P42345 - P23443 pathway accounted for Andro-induced autophagy . Finally , Andro-driven inhibition of the Q96P20 inflammasome and amelioration of murine models for colitis and CAC were significantly blocked by Q14457 knockdown , or by various autophagy inhibitors . Taken together , our findings demonstrate that mitophagy-mediated Q96P20 inflammasome inhibition by Andro is responsible for the prevention of CAC . Our data may help guide decisions regarding the use of Andro in patients with inflammatory bowel diseases , which ultimately reduces the risk of CAC . Estradiol increases cell growth in human astrocytoma cell lines through ERα activation and its interaction with Q15788 and Q9Y6Q9 coactivators . Estradiol ( E2 ) regulates several cellular functions through the interaction with estrogen receptor subtypes , ERα and ERβ , which present different functional and regulation properties . ER subtypes have been identified in human astrocytomas , the most common and aggressive primary brain tumors . We studied the role of ER subtypes in cell growth of two human astrocytoma cell lines derived from tumors of different evolution grades : U373 and D54 ( grades III and IV , respectively ) . E2 significantly increased the number of cells in both lines and the co-administration with an ER antagonist ( ICI 182 , 780 ) significantly blocked E2 effects . ERα was the predominant subtype in both cell lines . E2 and ICI 182 , 780 down-regulated ERα expression . The number of U373 and D54 cells significantly increased after PPT ( ERα agonist ) treatment but not after DPN ( ERβ agonist ) one . To determine the role of Q15788 and Q9Y6Q9 coactivators in ERα induced cell growth , we silenced them with RNA interference . Coactivator silencing blocked the increase in cell number induced by PPT . The content of proteins involved in proliferation and metastasis was also determined after PPT treatment . Western blot analysis showed that in U373 cells the content of PR isoforms ( PR-A and PR-B ) , P00533 , P15692 and cyclin D1 increased after PPT treatment while in D54 cells only the content of P00533 was increased . Our results demonstrate that E2 induces cell growth of human astrocytoma cell lines through ERα and its interaction with Q15788 and Q9Y6Q9 and also suggest differential roles of ERα on cell growth depending on astrocytoma grade . Salacia oblonga extract increases glucose transporter 4-mediated glucose uptake in Q9BTT4 rat myotubes : role of mangiferin . BACKGROUND AND AIMS : To evaluate if the antidiabetic properties of Salacia oblonga extract are mediated not only by inhibiting intestinal alpha-glycosidases but also by enhancing glucose transport in muscle and adipose cells . METHODS : S. oblonga extract effects on 2-deoxy-D-glucose uptake were assayed in muscle Q9BTT4 -myotubes and 3T3-adipocytes . In Q9BTT4 -myotubes , the amount and translocation of glucose transporters were assayed . A fractionation of the extract was carried out to identify the active compounds . Furthermore , we analyzed the phosphorylation status of key components of signaling pathways that are involved in the molecular mechanisms regulating glucose uptake . RESULTS : S. oblonga extract increased 2-deoxy-D-glucose uptake by 50 % in Q9BTT4 -myotubes and 3T3-adipocytes . In Q9BTT4 -myotubes , the extract increased up to a 100 % the P14672 content , activating P14672 promoter transcription and its translocation to the plasma membrane . Mangiferin was identified as the bioactive compound . Furthermore , mangiferin effects were concomitant with the phosphorylation of DB00131 -activated protein kinase without the activation of P31749 /Akt . The effect of mangiferin on 2-deoxy-D-glucose uptake was blocked by GW9662 , an irreversible P37231 antagonist . CONCLUSIONS : S. oblonga extract and mangiferin may exert their antidiabetic effect by increasing P14672 expression and translocation in muscle cells . These effects are probably mediated through two independent pathways that are related to DB00131 -activated protein kinase and P37231 . Common variants in O75369 / O75718 , not Q9NR81 at 3p , are associated with osteoporosis in southern Chinese women . SUMMARY : We performed an association study of five candidate genes within chromosome 3p14-25 in 1,080 Chinese female subjects . Polymorphisms in O75369 / O75718 are associated with bone mineral density ( BMD ) in Chinese . INTRODUCTION : Chromosomal region 3p14-25 has shown strong evidence of linkage to BMD in genome-wide linkage scans . The variants responsible for this linkage signal , nonetheless , remain obscure . METHODS : Thirty SNPs in five positional and functional candidate genes within 3p14-25 ( P37231 , O75718 , P13385 , Q03431 , and O75369 ) and rs7646054 in the Q9NR81 gene were genotyped in a case-control cohort of 1,080 Chinese females . Allelic and haplotypic association were tested using logistic regression analysis implemented in PLINK software . Potential transcription factor binding sites were predicted with MatInspector . RESULTS : Multiple SNPs and haplotypes in O75369 were significantly associated with BMDs , with the strongest association between lumbar spine BMD and rs9828717 ( p = 0.005 ) . SNP rs7623768 and the haplotype G-C of rs4076086-rs7623768 in O75718 were associated with femoral neck BMD ( p = 0.009 and p = 0.003 , respectively ) . Q03431 showed haplotypic associations with lumbar spine and femoral neck BMD ( p = 0.02 and p = 0.044 , respectively ) . Nevertheless , the association between rs7646054 in Q9NR81 and BMD observed in Caucasians was not replicated in our samples . Comparative genomics analysis indicated that rs9828717 is located within a highly conserved region . The minor T allele at rs9828717 may lead to loss of binding site for nuclear factor of activated T cells which binds and triggers the transcriptional program of osteoblasts . CONCLUSIONS : Our data suggest that variants in O75369 and O75718 at 3p are involved in BMD regulation in southern Chinese .

MH_train_1333

MH_train_1333

MH_train_1333

interacts_with DB09280?

multiple_choice

Critical role of P21453 and integrin β4 in P14210 /c- DB00134 -mediated increases in vascular integrity . Vascular endothelial cell ( EC ) barrier integrity is critical to vessel homeostasis whereas barrier dysfunction is a key feature of inflammatory disorders and tumor angiogenesis . We previously reported that hepatocyte growth factor ( P14210 ) -mediated increases in EC barrier integrity are signaled through a dynamic complex present in lipid rafts involving its receptor , c- DB00134 . We extended these observations to confirm that P21453 ( sphingosine 1-phosphate receptor 1 ) and integrin β4 ( P16144 ) are essential participants in P14210 -induced EC barrier enhancement . Immunoprecipitation experiments demonstrated P14210 -mediated recruitment of c- DB00134 , P16144 and P21453 to caveolin-enriched lipid rafts in human lung EC with direct interactions of c- DB00134 with both P21453 and P16144 accompanied by c- DB00134 -dependent P21453 and P16144 transactivation . Reduced P21453 expression ( siRNA ) attenuated both P16144 and Rac1 activation as well as c- DB00134 / P16144 interaction and resulted in decreased transendothelial electrical resistance . Furthermore , reduced P16144 expression attenuated P14210 -induced c- DB00134 activation , c- DB00134 / P21453 interaction , and effected decreases in Q14703 - and P14210 -induced EC barrier enhancement . Finally , the c- DB00134 inhibitor , DB05030 , suppressed P14210 -induced c- DB00134 activation as well as P21453 and P16144 transactivation . These results support a critical role for P21453 and P16144 transactivation as rate-limiting events in the transduction of P14210 signals via a dynamic c- DB00134 complex resulting in enhanced EC barrier integrity . Genomic landscape of non-small cell lung cancer in smokers and never-smokers . We report the results of whole-genome and transcriptome sequencing of tumor and adjacent normal tissue samples from 17 patients with non-small cell lung carcinoma ( NSCLC ) . We identified 3,726 point mutations and more than 90 indels in the coding sequence , with an average mutation frequency more than 10-fold higher in smokers than in never-smokers . Novel alterations in genes involved in chromatin modification and DNA repair pathways were identified , along with Q9UI36 , P13569 , P78509 , Q2M3G0 , and P14210 . Deep digital sequencing revealed diverse clonality patterns in both never-smokers and smokers . All validated EFGR and P01116 mutations were present in the founder clones , suggesting possible roles in cancer initiation . Analysis revealed 14 fusions , including P08922 and Q9UM73 , as well as novel metabolic enzymes . Cell-cycle and JAK- P35610 pathways are significantly altered in lung cancer , along with perturbations in 54 genes that are potentially targetable with currently available drugs . [ Molecular-genetic analysis of certain mutations of the " cystic fibrosis gene " in Moldavia. Characteristics of molecular markers and their linkage with various mutations ] . Sixty-one patients with cystic fibrosis ( CF ) from Moldova were tested for mutations delta F508 , G551D , and R553X . Frequencies of various alleles of the repeated GATT sequence in intron 6B of the P13569 gene , their linkage to other polymorphic markers , and various mutations were determined . The frequency of occurrence of mutation delta F508 was only 25 % . An absolute majority of CF patients ( 80 % ) had pancreatic insufficiency . Mutations G551D and R553X were not found in our sample . Each of 31 chromosomes with mutation delta F508 carries the 6-GATT allele . Most " non delta F508 " ( 78 % ) and normal ( 80 % ) chromosomes were marked by 7-GATT allele . Twenty-seven delta F508 chromosomes ( 96.4 % ) belong to haplotype B6 , and only one to O00590 . Most chromosomes with " non delta F508 " mutations are associated with haplotypes D7 ( 26.3 % ) and P10643 ( 21 % ) . In addition , a significant portion of chromosomes from this subgroup were associated with haplotypes A7 ( 23.7 % ) , A6 ( 10.5 % ) , and P13671 ( 2.7 % ) , which are not yet described for mutant chromosomes . The results obtained demonstrate that CF in Moldova is mainly associated with mutations other than delta F508 , G551D , and R553X . Severe forms of the disease , with pancreatic insufficiency , are more frequently caused by these mutations ; moreover , our data provides strong evidence about the presence of at least seven additional CF mutations in Moldova , apart from delta F508 , G551D , and R553X . Some of these are probably not described . The association between thrombophilic gene mutations and recurrent pregnancy loss . PURPOSE : To determine whether the Factor V ( 1691G/A ) , Factor V HR2 ( 4070A/G ) , P00734 ( 20210G/A ) , P05121 ( -675 I/D , 5G/4G ) , P12821 ( intron 16 I/D ) , Factor VII ( Gln353Arg ) , Factor XIII ( Val34Leu ) , β-fibrinogen ( -455G/A ) , Glycoprotein Ia ( 807C/T ) , tPA ( intron 8 D/I ) gene mutations could be risk factors for recurrent pregnancy loss ( RPL ) . METHODS : Genotyping of thrombophilic gene mutations were carried out by amplification Refractory Mutation System-PCR ( Q9ULH0 -PCR ) method after DNA extraction . RESULTS : We found that the mutant allele frequencies of Factor V ( 1691G/A ) , Factor V HR2 ( 4070A/G ) , P00734 ( 20210G/A ) , P05121 ( -675 I/D , 5G/4G ) , Factor XIII ( Val34Leu ) and β-fibrinogen ( -455G/A ) were more seen in the case group compared with the healthy control ; However , the difference between the two group is not statistically significant ( p > 0.05 ) . Whilst the mutant allele frequencies of other studied genes were lower in the case in comparison to the fertile control women ( p > 0.05 ) . CONCLUSION : Taken together , our data has shown that the prevalence of thrombophilic gene mutations was similar in women with RPL and healthy controls . Therefore , it appears that further studies on large-scale population and other genetic variants will be needed to conclusively find candidate genes for RPL unknown etiology in the future . Genetic analysis of 56 polymorphisms in 17 genes involved in methionine metabolism in patients with abdominal aortic aneurysm . BACKGROUND : Previous studies suggested an association between abdominal aortic aneurysm ( AAA ) and hyperhomocysteinaemia , a complex trait determined by genetic and environmental factors . Our hypothesis was that polymorphisms in genes directly or indirectly involved in methionine metabolism may contribute to AAA susceptibility . METHOD : We studied 56 polymorphisms in P42898 , Q99707 , Q9UBK8 , P35520 , P11586 , P41440 , P40261 , P20062 , P23526 , Q93088 , Q9H2M3 , Q04609 , P04818 , Q7L5Y1 , P34896 , P27169 , Q15165 genes according to their demonstrated/putative function , localisation in promoter or regulatory and coding regions and/or heterozygosity values > 0.300 . Polymorphisms were evaluated by using a primer extension based microarray technology in 423 AAA patients and 423 matched controls . RESULTS : All polymorphisms resulted in Hardy-Weinberg equilibrium in patients and controls . At the multiple logistic regression analysis adjusted for traditional cardiovascular risk factors ( sex , age , hypertension , smoking habit , dyslipidaemia , diabetes ) and chronic obstructive pulmonary disease ( P48444 ) , rs8003379 P11586 ( odds ratio ( OR ) 0.41 , 95 % confidence interval ( CI ) 0.26 to 0.65 ) and rs326118 Q9UBK8 ( OR 0.47 , 95 % CI 0.29 to 0.77 ) polymorphisms resulted in independent susceptibility factor for AAA . CONCLUSIONS : After haplotype reconstruction , logistic regression analyses adjusted for traditional risk factors and P48444 showed a significant association among AAA and P23526 , Q04609 , P11586 , Q99707 , P40261 , P27169 and P04818 haplotypes . Our findings offer new insights into the pathogenesis of AAA . Genetics of idiopathic disseminated bronchiectasis . Bronchiectasis is an abnormal dilation of bronchi , consequent to the destruction of their walls . It is included in the category of obstructive pulmonary diseases , along with chronic obstructive pulmonary disease ( P48444 ) , asthma , and cystic fibrosis . In approximately 50 % of cases , bronchiectasis is associated with underlying conditions ; in the remainder , known causes are not ascertainable ( idiopathic bronchiectasis ) . A search for genetic determinants of this phenotype , with the cystic fibrosis gene as a candidate , has been performed by three independent groups . The results of this search agreed on the association of bronchiectasis with cystic fibrosis gene mutations and polymorphisms . The cystic fibrosis gene is also associated with bronchiectasis due to rheumatoid arthritis and allergic bronchopulmonary aspergillosis . A few other genes have been investigated in idiopathic bronchiectasis , with negative results . Idiopathic bronchiectasis is , therefore , to be considered as an obstructive multifactorial disorder belonging to the category of cystic fibrosis monosymptomatic diseases ( or P13569 -opathies ) , whose pathogenesis is influenced by environmental factors and other undetermined genes . A novel tyrosine kinase switch is a mechanism of imatinib resistance in gastrointestinal stromal tumors . P10721 or alpha-platelet-derived growth factor receptor ( alpha- P09619 ) activating mutations are the pathogenic mechanisms that characterize gastrointestinal stromal tumors ( GIST ) . Despite excellent responses to imatinib mesylate ( IM ) , patients are relapsing . We developed an IM-resistant GIST cell line ( GIST-R ) from the IM-sensitive GIST882 cell line ( GIST-S ) by growing these cells in IM . Gene expression profiling ( GEP ) of GIST-S , GIST-R cells and two IM resistant GIST patients demonstrated that P10721 is downregulated implying a major role in IM resistance . Instead , GIST-R cells have acquired IM resistance by overexpressing the oncogenic receptor tyrosine kinase - P30530 - in a ' kinase switch ' . Further , the two IM resistant GIST patients express P30530 and not c-Kit , seen by immunohistochemistry ( IHC ) . Real time reverse transcriptase-polymerase chain reaction and Western blotting of the GIST-S and GIST-R cells confirmed the switch from Kit to P30530 . In GIST-R , P30530 is tyrosine phosphorylated and its ligand growth-arrest-specific gene 6 is overexpressed implying autocrine activation . The kinase switch is associated with a morphological change from spindle to epithelioid . Molecular modeling of the kinase domain of mutant c-Kit ( V654A ) and P30530 showed no binding to IM but efficient binding to DB05216 , a novel c-Kit/ P30530 kinase inhibitor . DB05216 synergizes with docetaxel ( taxotere ) and is cytotoxic to GIST cells . P03372 ( ER ) expression and function in the pregnant human myometrium : estradiol via ERα activates P27361 /2 signaling in term myometrium . DB00286 are thought to promote labor by increasing the expression of pro-contraction genes in myometrial cells . The specific estrogen receptors ( ( ERs : ERα and ERβ ( also known as P03372 and Q92731 ) ) and G protein-coupled receptor 30 ( Q99527 ; also known as Q99527 ) ) and signaling pathways that mediate these actions are not clearly understood . In this study , we identified the ERs expressed in the pregnant human myometrium and determined a key extranuclear signaling pathway through which estradiol ( E(2) ) modulates expression of the gene encoding the oxytocin receptor ( P30559 ) , a major pro-contraction protein . Using quantitative RT-PCR , we found that ERα and Q99527 mRNAs were expressed in the human pregnant myometrium while ERβ mRNA was virtually undetectable . While mRNA encoding ERα was the predominant ER transcript in the pregnant myometrium , ERα protein was largely undetectable in myometrial tissue by immunoblotting . Pharmacological inhibition of 26S proteasome activity increased ERα protein abundance to detectable levels in term myometrial explants , however , indicating rapid turnover of ERα protein by proteasomal processing in the pregnant myometrium . E(2) stimulated rapid extranuclear signaling in myometrial explants , as evidenced by increased extracellularly regulated kinase ( P27361 /2 ) phosphorylation within 10 min . This effect was inhibited by pre-treatment with an ER antagonist , ICI 182 780 , indicating the involvement of ERα . Inhibition of P29323 signaling abrogated the ability of E(2) to stimulate P30559 gene expression in myometrial explants . We conclude that estrogenic actions in the human myometrium during pregnancy , including the stimulation of contraction-associated gene expression , can be mediated by extranuclear signaling through ERα via activation of the P29323 /mitogen-activated protein kinase pathway . Functional methionine synthase deficiency ( cblE and cblG ) : clinical and biochemical heterogeneity . Functional methionine synthase deficiency is generally characterized by homocystinuria and hypomethioninemia in the absence of methylmalonic aciduria . Patients are divided into two classes , cblE and cblG , on the basis of complementation analysis . Presentation has usually been in the first 2 years of life , but one patient came to medical attention at age 21 years with symptoms initially diagnosed as multiple sclerosis . Common findings among 11 patients ( 4 with cblE and 7 with cblG ) have included megaloblastic anemia ( all patients ) and various neurological deficits including developmental retardation ( 10 patients ) , cerebral atrophy ( 8 patients ) , hypotonia ( 7 patients ) , EEG abnormalities ( 6 patients ) , and nystagmus ( 5 patients ) . Hypertonia , seizures , blindness , and ataxia were less frequent . All patients have responded to therapy with cobalamin with resolution of anemia and biochemical abnormalities ; neurological deficits resolved more slowly and in some cases incompletely . DB00200 has been more effective than cyanocobalamin . Fibroblasts from patients with cblE ( 5 patients ) and cblG ( 6 patients ) all showed decreased intracellular levels of methylcobalamin ( DB03614 ) and decreased incorporation of label from 5-methyltetrahydrofolate into macromolecules , suggesting decreased activity of the DB03614 -dependent enzyme methionine synthase . Q99707 specific activity in extracts of all cblE fibroblasts was normal or near-normal under standard reducing conditions ; synthase specific activity in extracts of 5 cblG patients was low but was high in a 6th patient measured in another laboratory . Thus , there is heterogeneity among patients with functional methionine synthase deficiency both in clinical presentation and in the results of biochemical studies of cultured cells . Crystal structure of the PDZ1 domain of human Na(+)/H(+) exchanger regulatory factor provides insights into the mechanism of carboxyl-terminal leucine recognition by class I PDZ domains . The Na(+)/H(+) exchanger regulatory factor ( O14745 ; also known as O14745 ) contains two PDZ domains that mediate the assembly of transmembrane and cytosolic proteins into functional signal transduction complexes . The O14745 PDZ1 domain interacts specifically with the motifs DSLL , DSFL , and DTRL present at the carboxyl termini of the beta(2) adrenergic receptor ( beta(2)AR ) , the platelet-derived growth factor receptor ( P09619 ) , and the cystic fibrosis transmembrane conductance regulator ( P13569 ) , respectively , and plays a central role in the physiological regulation of these proteins . The crystal structure of the human O14745 PDZ1 has been determined at 1.5 A resolution using multiwavelength anomalous diffraction phasing . The overall structure is similar to known PDZ structures , with notable differences in the O14745 PDZ1 carboxylate-binding loop that contains the GYGF motif , and the variable loop between the beta2 and beta3 strands . In the crystalline state , the carboxyl-terminal sequence DEQL of PDZ1 occupies the peptide-binding pocket of a neighboring PDZ1 molecule related by 2-fold crystallographic symmetry . This structure reveals the molecular mechanism of carboxyl-terminal leucine recognition by class I PDZ domains , and provides insights into the specificity of O14745 interaction with the carboxyl termini of several membrane receptors and ion channels , including the beta(2)AR , P09619 , and P13569 . Multiple endocytic signals in the C-terminal tail of the cystic fibrosis transmembrane conductance regulator . The cystic fibrosis transmembrane conductance regulator ( P13569 ) is a DB02527 -dependent protein kinase ( PKA ) -activated chloride channel that is localized to the plasma membrane and endosomal compartment . Endosomal targeting of P13569 is attributed to the DB00135 (1424)-based internalization signal , identified in the C-terminal tail of the channel . Mutation of the DB00135 (1424) residue could partly inhibit the endocytosis of P13569 and its association with the adapter protein P05549 . To reveal additional endosomal targeting signals , site-directed mutagenesis of both a chimaera , composed of a truncated form of interleukin 2 receptor alpha chain ( TacT ) and the C-terminal tail of P13569 ( Ct ) , and the full-length P13569 was performed . Morphological and functional assays revealed the presence of multiple internalization motifs at the C-terminus , consisting of a phenylalanine-based motif ( DB00120 (1413) ) and a bipartite endocytic signal , comprising a tyrosine ( DB00135 (1424) ) and a di- DB00149 -based ( DB00149 (1430)- DB00149 ) motif . Whereas the replacement of any one of the three internalization motifs with alanine prevented the endocytosis of the TacT-Ct chimaera , mutagenesis of DB00120 (1413)- DB00149 impaired the biosynthetic processing of P13569 , indicating that DB00120 (1413) is indispensable for the native structure of P13569 . In contrast , replacement of DB00149 (1430)- DB00149 - and DB00135 (1424)-based signals with alanine increased the cell-surface density of both the chimaeras and P13569 in an additive manner . These results suggest that the internalization of P13569 is regulated by multiple endocytic sorting signals . Effects on thrombin generation of single injections of DB02351 in patients with calf vein thrombosis . STUDY OBJECTIVE : To determine whether single injections of DB02351 , a direct thrombin inhibitor , can inhibit thrombin generation in patients with calf vein thrombosis and , if so , if the inhibition is sustained . DESIGN : Phase II open label cohort study . SETTING : Tertiary-care referral centres , university affiliated hospitals . PATIENTS : 10 patients with venographically-demonstrated calf vein thrombosis . INTERVENTION : Patients received a single injection of DB02351 , either 1.0 mg/kg subcutaneously or 0.6 mg/kg as a 15 min intravenous infusion . P00734 fragment ( F1++2 ) levels , as an index of thrombin generation , were measured before as well as 6 h post- and 24 h post- DB02351 administration . Patients were followed with non-invasive tests to detect thrombus extension into the proximal veins . RESULTS : There was a significant reduction in the levels of F1+2 with both regimens , 6 h after DB02351 . The F1+2 levels 24 h post- DB02351 showed a significant increase relative to the 6 h post- DB02351 results . One patient developed thrombus extension into the popliteal vein and was treated with conventional anticoagulants . CONCLUSION : The single injections of DB02351 used in the study produced incomplete and temporary suppression of F1+2 . Complete and permanent inhibition of thrombin generation with DB02351 in patients with calf vein thrombosis may require higher doses , multiple subcutaneous injections and/or prolonged intravenous infusion . Transporters in cholelithiasis . Gallstones are a common and costly disease with a projected increase in prevalence due to the increasing ageing population . Numerous endogenous and environmental factors are aetiologically related to this multifactorial disease , and genetic studies continue to unravel the pathobiological mechanisms related to gallstone formation . In particular , variants of genes encoding hepatobiliary transporters have been implicated in gallstone disease and , given their ability to influence biliary lipid composition , have undergone considerable investigation . Here we summarize the role of enterohepatic transporters in cholelithogenesis with a particular focus on pertinent DB00171 -binding cassette transporters ( P21439 , O95342 , P13569 , and Q9H222 / Q9UBA6 ) . Genomic organization and partial duplication of the human alpha7 neuronal nicotinic acetylcholine receptor gene ( P36544 ) . The human alpha7 neuronal nicotinic acetylcholine receptor gene ( HGMW-approved symbol P36544 ) has been characterized from genomic clones . The gene is similar in structure to the chick alpha7 gene with 10 exons and conserved splice junction positions . The size of the human gene is estimated to be larger than 75 kb . A putative promoter 5' of the translation start in exon 1 has been cloned and sequenced . The promoter region lacks a TATA box and has a high GC content ( 77 % ) . Consensus Sp1 , P05549 , Egr-1 , and CREB transcription factor binding sites appear to be conserved between bovine and human genes . The alpha7 nAChR gene was found to be partially duplicated , with both loci mapping to the chromosome 15q13 region . A yeast artificial chromosome contig was constructed over a genetic distance of 5 cM that includes both alpha7 loci and the region between them . Four novel exons are described , located in genomic clones containing the partially duplicated gene . The duplicated sequences , including the novel exons , are expressed in human brain . P01375 polymorphisms as a potential modifier gene in the cystic fibrosis . Modifier genes , as the P01375 -α gene , can modulate the cystic fibrosis ( CF ) severity . Thus , -238G > A and -308G > A polymorphisms of P01375 -α gene were analyzed as modifiers of CF . In this context , the present study enrolled 49 CF patients ( diagnosis performed by sweat test and complete P13569 screening ) . The -238G > A polymorphism analysis was performed by Q9ULH0 -PCR , and -308G > A , by PCR-RFLP . In our data , the -238G > A polymorphism was not associated with clinical variability . The AA genotype for -308G > A polymorphism was a risk factor for early gastrointestinal symptoms ( OR=5.98 , 95 % CI=1.06-49.68 ) and protection for the first Pseudomonas aeruginosa ( OR=0.05 , 95 % CI=0.0003-0.007 ) . For the first P. aeruginosa , GA genotype was a risk factor ( OR=10.2 , 95 % CI=1.86-84.09 ) ; for the same genotype , the diagnosis was made in minor time than the AA genotype ( p=0.031 ) . Considering the -308G > A polymorphism alleles , the G allele was a risk factor for early pulmonary symptoms ( OR=3.81 , 95 % CI=1.13-12.97 ) and P. aeruginosa ( OR=66.77 , 95 % CI=15.18-482.7 ) ; however , the same allele showed better transcutaneous oxygen saturation ( OR=9.24 , 95 % CI=1.53-206.1 ) . The A allele was a protective factor for early pulmonary symptoms ( OR=12.26 , 95 % CI=0.08-0.89 ) and P. aeruginosa ( OR=12.15 , 95 % CI=0002-0007 ) , however , the same allele was a risk factor for worst transcutaneous oxygen saturation ( OR=7.01 , 95 % CI=1.14-157.4 ) . As conclusion , the -308G > A polymorphism of the P01375 -α gene was associated with the CF severity . Potentiator ivacaftor abrogates pharmacological correction of ΔF508 P13569 in cystic fibrosis . Cystic fibrosis ( CF ) is caused by mutations in the CF transmembrane conductance regulator ( P13569 ) . Newly developed " correctors " such as DB09280 ( VX-809 ) that improve P13569 maturation and trafficking and " potentiators " such as ivacaftor ( VX-770 ) that enhance channel activity may provide important advances in CF therapy . Although VX-770 has demonstrated substantial clinical efficacy in the small subset of patients with a mutation ( G551D ) that affects only channel activity , a single compound is not sufficient to treat patients with the more common P13569 mutation , ΔF508 . Thus , patients with ΔF508 will likely require treatment with both correctors and potentiators to achieve clinical benefit . However , whereas the effectiveness of acute treatment with this drug combination has been demonstrated in vitro , the impact of chronic therapy has not been established . In studies of human primary airway epithelial cells , we found that both acute and chronic treatment with VX-770 improved P13569 function in cells with the G551D mutation , consistent with clinical studies . In contrast , chronic VX-770 administration caused a dose-dependent reversal of VX-809-mediated P13569 correction in ΔF508 homozygous cultures . This result reflected the destabilization of corrected ΔF508 P13569 by VX-770 , markedly increasing its turnover rate . Chronic VX-770 treatment also reduced mature wild-type P13569 levels and function . These findings demonstrate that chronic treatment with P13569 potentiators and correctors may have unexpected effects that can not be predicted from short-term studies . Combining these drugs to maximize rescue of ΔF508 P13569 may require changes in dosing and/or development of new potentiator compounds that do not interfere with P13569 stability . Molecular mechanisms governing different pharmacokinetics of ginsenosides and potential for ginsenoside-perpetrated herb-drug interactions on Q9NPD5 . BACKGROUND AND PURPOSE : Ginsenosides are bioactive saponins derived from Panax notoginseng roots ( Sanqi ) and ginseng . Here , the molecular mechanisms governing differential pharmacokinetics of 20(S)-protopanaxatriol-type ginsenoside Rg1 , ginsenoside Re and notoginsenoside Q96GN5 and 20(S)-protopanaxadiol-type ginsenosides Rb1 , Rc and Rd were elucidated . EXPERIMENTAL APPROACH : Interactions of ginsenosides with human and rat hepatobiliary transporters were characterized at the cellular and vesicular levels . A rifampin-based inhibition study in rats evaluated the in vivo role of organic anion-transporting polypeptide (Oatp)1b2 . Plasma protein binding was assessed by equilibrium dialysis . Drug-drug interaction indices were calculated to estimate potential for clinically relevant ginsenoside-mediated interactions due to inhibition of human OATP1Bs . KEY RESULTS : All the ginsenosides were bound to human Q9NPD5 and rat Oatp1b2 but only the 20(S)-protopanaxatriol-type ginsenosides were transported . Human multidrug resistance-associated protein (MRP)2/breast cancer resistance protein ( Q9UNQ0 ) /bile salt export pump ( O95342 ) /multidrug resistance protein-1 and rat Mrp2/Bcrp/Bsep also mediated the transport of the 20(S)-protopanaxatriol-type ginsenosides . Glomerular-filtration-based renal excretion of the 20(S)-protopanaxatriol-type ginsenosides was greater than that of the 20(S)-protopanaxadiol-type counterparts due to differences in plasma protein binding . DB01045 -impaired hepatobiliary excretion of the 20(S)-protopanaxatriol-type ginsenosides was effectively compensated by the renal excretion in rats . The 20(S)-protopanaxadiol-type ginsenosides were potent inhibitors of Q9NPD5 . CONCLUSION AND IMPLICATIONS : Differences in hepatobiliary and in renal excretory clearances caused markedly different systemic exposure and different elimination kinetics between the two types of ginsenosides . Caution should be exercised with the long-circulating 20(S)-protopanaxadiol-type ginsenosides as they could induce hepatobiliary herb-drug interactions , particularly when patients receive long-term therapies with high-dose i.v. Sanqi or ginseng extracts . Influence of cystic fibrosis transmembrane conductance regulator on gene expression in response to Pseudomonas aeruginosa infection of human bronchial epithelial cells . Chronic lung infection by Pseudomonas aeruginosa causes significant morbidity in cystic fibrosis patients initiated by the failure of innate immune responses . We used microarray analysis and real-time PCR to detect transcriptional changes associated with cytokine production in isogenic bronchial epithelial cell lines with either wild-type ( WT ) or mutant cystic fibrosis transmembrane conductance regulator ( P13569 ) in response to P. aeruginosa infection . The transcription of four NF-kappaB-regulated cytokine genes was maximal in the presence of WT P13569 : the interleukin-8 ( P10145 ) , P05231 , P09341 , and intracellular adhesion molecule 1 ( P05362 ) genes . Analysis of protein expression in two cell lines paired for wild-type and mutant P13569 with three P. aeruginosa strains showed P05231 and P10145 expressions were consistently enhanced by the presence of WT P13569 in both cell lines with all three strains of P. aeruginosa , although some strains gave small P10145 increases in cells with mutant P13569 . P09341 production showed consistent enhancement in cells with WT P13569 using all three bacterial strains in one cell line , whereas in the other cell line , P09341 showed a significant increase in cells with either WT or mutant P13569 . P05362 was unchanged at the protein level in one of the cell lines but did show mild enhancement with WT P13569 in the other cell pair . Inhibitions of NF-kappaB prior to infection indicated differing degrees of dependence on NF-kappaB for production of the cytokines , contingent on the cell line . Cytokine effectors of innate immunity to P. aeruginosa were found to be positively influenced by the presence of WT P13569 , indicating a role in resistance to P. aeruginosa infection . Differential regulation of cystic fibrosis transmembrane conductance regulator by interferon gamma in mast cells and epithelial cells . P13569 ( P13569 ) is a P13569 in epithelial cells ; recently , we identified it in mast cells . Previous work that we confirmed showed that interferon gamma ( IFNgamma ) down-regulated P13569 expression in epithelial cells ( T84 ) , but by contrast , we found that IFNgamma up-regulated P13569 mRNA and protein expression in rat and human mast cells . IFNgamma up-regulation of P13569 in mast cells was inhibited by p38 and extracellular signal-regulated kinase ( P29323 ) kinase inhibitors but not a Janus tyrosine kinase (JAK)2 inhibitor , whereas in T84 cells IFNgamma-mediated down-regulation of P13569 was O60674 -dependent and P29323 - and p38-independent . Furthermore , IFNgamma down-regulation of P13569 in T84 epithelial cells was P42224 -dependent , but up-regulation of P13569 in mast cells was P42224 -independent . Thus , differential regulatory pathways of P13569 expression in mast cells and epithelial cells exist that depend upon either p38/ P29323 or JAK/ P35610 pathways , respectively . Surprisingly , IFNgamma treatment of mast cells inhibited Cl(-) efflux , in contrast to up-regulation of P13569 /mRNA and protein expression . However , down-regulation of Cl(-) flux correlated with IFNgamma-mediated inhibition of mediator secretion . This and other work suggests that the effect of IFNgamma on P13569 expression in mast cells is important for their function . Effects of an alpha 7-nicotinic agonist on default network activity in schizophrenia . BACKGROUND : 3-(2,4-dimethoxybenzylidene)-anabaseine ( DB05708 ) is a partial agonist at α7 nicotinic acetylcholine receptors that has been evaluated clinically for treatment of schizophrenia . This study examined the effects of DB05708 on default network activity as a biomarker for drug effects on pathologic brain function associated with schizophrenia . METHODS : Placebo and two doses of DB05708 were administered in a random , double-blind crossover design during a Phase 2 study of DB05708 . Functional magnetic resonance imaging was performed on 16 nonsmoking patients with schizophrenia while they performed a simple eye movement task . Independent component analysis was used to identify the default network component . Default network changes were evaluated in the context of a polymorphism in P36544 , the α7-nicotinic acetylcholine receptor subunit gene , which was previously found to be associated with schizophrenia . RESULTS : Compared with placebo , both 150 and 75 mg twice daily DB05708 altered default network activity , including a reduction in posterior cingulate , inferior parietal cortex , and medial frontal gyrus activity and an increase in precuneus activity . The most robust difference , posterior cingulate activity reduction , was affected by P36544 genotype . CONCLUSIONS : The observed DB05708 -related changes are consistent with improved default network function in schizophrenia . Pharmacogenetic analysis indicates mediation of the effect through the α7-nicotinic receptor . These results further implicate nicotinic cholinergic dysfunction in the disease and suggest that default network activity may be a useful indicator of biological effects of novel therapeutic agents . Molecular basis for the immunostimulatory activity of guanine nucleoside analogs : activation of Q9NYK1 . Certain Q99618 -substituted and N7 , Q99618 -disubstituted guanine ribonucleosides comprise a class of small molecules with immunostimulatory activity . In a variety of animal models , these agents stimulate both humoral and cellular immune responses . The antiviral actions of these guanosine analogs have been attributed to their ability to induce type I IFNs . However , the molecular mechanisms by which the guanosine analogs potentiate immune responses are not known . Here , we report that several guanosine analogs activate Q9NYK1 ( Q9NYK1 ) . DB04860 , 7-deazaguanosine , and related guanosine analogs activated mouse immune cells in a manner analogous to known TLR ligands , inducing cytokine production in mouse splenocytes ( P05231 and IL-12 , type I and II IFNs ) , bone marrow-derived macrophages ( P05231 and IL-12 ) , and in human peripheral blood leukocytes ( type I IFNs , tumor necrosis factor alpha and IL-12 ) . The guanosine congeners also up-regulated costimulatory molecules and MHC I/II in dendritic cells . Genetic complementation studies in human embryonic kidney 293 cells confirmed that the guanosine analogs activate cells exclusively via Q9NYK1 . The stimulation of Q9NYK1 by the guanosine analogs in human cells appears to require endosomal maturation because inhibition of this process with chloroquine significantly reduced the downstream activation of NF-kappaB . However , Q9NR97 activation by R-848 and O60603 activation by [ S-[2,3-bis(palmitoyloxy)-(2-RS)-propyl]-N-palmitoyl-R- DB00151 -S- DB00133 -Lys4-OH , trihydrochloride ) ] were not inhibited by chloroquine , whereas Q9NR96 activation by CpG oligodeoxynucleotides was abolished . In summary , we present evidence that guanosine analogs activate immune cells via Q9NYK1 by a pathway that requires endosomal maturation . Thus , the B cell-stimulating and antiviral activities of the guanosine analogs may be explained by their Q9NYK1 -activating capacity . Electrostatic steering at acetylcholine binding sites . The electrostatic environments near the acetylcholine binding sites on the nicotinic acetylcholine receptor ( nAChR ) and acetylcholinesterase were measured by diffusion-enhanced fluorescence energy transfer ( DEFET ) to determine the influence of long-range electrostatic interactions on ligand binding kinetics and net binding energy . Changes in DEFET from variously charged Tb3+ -chelates revealed net potentials of -20 mV at the nAChR agonist sites and -14 mV at the entrance to the P22303 active site , in physiological ionic strength conditions . The potential at the alphadelta-binding site of the nAChR was determined independently in the presence of DB01199 to be -14 mV ; the calculated potential at the alphagamma-site was approximately threefold stronger than at the alphadelta-site . By determining the local potential in increasing ionic strength , Debye-Hückel theory predicted that the potentials near the nAChR agonist binding sites are constituted by one to three charges in close proximity to the binding site . Examination of the binding kinetics of the fluorescent acetylcholine analog dansyl- P13671 -choline at ionic strengths from 12.5 to 400 mM revealed a twofold decrease in association rate . Debye-Hückel analysis of the kinetics revealed a similar charge distribution as seen by changes in the potentials . To determine whether the experimentally determined potentials are reflected by continuum electrostatics calculations , solutions to the nonlinear Poisson-Boltzmann equation were used to compute the potentials expected from DEFET measurements from high-resolution models of the nAChR and P22303 . These calculations are in good agreement with the DEFET measurements for P22303 and for the alphagamma-site of the nAChR . We conclude that long-range electrostatic interactions contribute -0.3 and -1 kcal/mol to the binding energy at the nAChR alphadelta- and alphagamma-sites due to an increase in association rates .

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A double-blind , placebo-controlled outpatient trial of pergolide for cocaine dependence . Results of preclinical studies suggest that pergolide , a mixed D(1)/ P14416 agonist , may be useful in treating cocaine dependence . To empirically investigate this possibility , we conducted a 5-year , double-blind , placebo-controlled clinical trial of two doses of pergolide ( 0.05 and 0.25 mg bid ) in subjects with cocaine dependence and combined cocaine/alcohol dependence . Data analysis was performed on an intent to treat population ( N=358 ) and a per protocol population ( N=108 ) with urine drug screens ( UDS ) used as the main outcome measure . There were no significant effects on UDS at either pergolide dose . DB01186 had no significant effect on alcohol use in the comorbid alcohol/cocaine dependence group . DB01186 does not appear to have clinical value in the treatment of cocaine dependence or in decreasing alcohol use in cocaine-dependent individuals at the presently studied doses . Agonism at P41595 receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5-hydroxytryptamine2B ( P41595 ) receptors . To evaluate whether agonism at P41595 receptors is a phenomenon of the class of the ergolines , we studied P41595 receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC50 8.42 and 8.72 ) . DB01200 acted as a partial agonist ( pEC50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5-HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 receptors seems not to be a class effect of the ergolines . O75469 induces Q02318 and regulates cholesterol metabolism in the intestine . Mitochondrial sterol 27-hydroxylase ( Q02318 ) catalyzes oxidative cleavage of the sterol side chain in the bile acid biosynthetic pathway in the liver and 27-hydroxylation of cholesterol in most tissues . Recent studies suggest that 27-hydroxycholesterol ( 27-HOC ) activates liver orphan receptor alpha ( LXRalpha ) and induces the cholesterol efflux transporters O95477 and P45844 in macrophages . The steroid- and bile acid-activated pregnane X receptor ( O75469 ) plays critical roles in the detoxification of bile acids , cholesterol metabolites , and xenobiotics . The role of Q02318 in the intestine is not known . This study investigated O75469 and Q02318 regulation of cholesterol metabolism in the human intestinal cell lines Caco2 and Ls174T . A human O75469 ligand , rifampicin , induced Q02318 mRNA expression in intestine cells but not in liver cells . DB01045 induced Q02318 gene transcription , increased intracellular 27-HOC levels , and induced O95477 and P45844 mRNA expression only in intestine cells . A functional O75469 binding site was identified in the human Q02318 gene . Chromatin immunoprecipitation assays revealed that rifampicin induced the O75469 recruitment of steroid receptor coactivator 1 to Q02318 chromatin . DB04540 loading markedly increased intracellular 27-HOC levels in intestine cells . DB01045 , 27-HOC , and a potent LXRalpha agonist , T0901317 , induced O95477 and P45844 protein expression and stimulated cholesterol efflux from intestine cells to apolipoprotein A-I and HDL . This study suggests an intestine-specific O75469 / Q02318 /LXRalpha pathway that regulates intestine cholesterol efflux and HDL assembly . P10275 regulation of P55008 cyclin and cyclin-dependent kinase function in the CWR22 human prostate cancer xenograft . Human prostate cancer is initially dependent on androgens for growth , and androgen-dependent cells undergo apoptosis after castration . However , a subset of androgen-responsive cells survives and eventually proliferates in the absence of testicular androgen . The high levels of androgen receptor in both androgen-dependent and recurrent tumors led us to investigate androgen regulation of cell cycle proteins in human prostate cancer using the CWR22 xenograft . Cellular proliferation decreased dramatically in CWR22 tumors after castration . DB00624 propionate ( TP ) treatment of castrated mice restored cellular proliferation after 24-48 hours . Growth of CWR22 tumors in the absence of testicular androgen recurred several months after castration . P06493 and P24941 , and cyclin A and cyclin B1 messenger RNAs were decreased 6 days after castration , increased 6-12 hours after TP treatment , and were expressed at high levels in recurrent CWR22 tumors . Coimmunoprecipitated cyclin B1/ P06493 and cyclin D1/ P11802 protein complexes decreased after castration and increased after TP treatment of castrated mice . In addition , P06493 and P24941 kinase activities were upregulated by androgen in parallel with hyperphosphorylation of retinoblastoma ( Rb ) protein . Despite the absence of testicular androgen in recurrent CWR22 , the levels of these androgen-regulated cyclin/ CDK protein complexes and hyperphosphorylation of Rb were equal to or greater than in tumors from intact mice . The results indicate that androgen receptor regulates cellular proliferation by control of CDK and cyclins at the transcriptional level and by post-translational modifications that influence cell cycle protein activity . DB06268 sodium for pulmonary hypertension . DB06268 is the first oral endothelin receptor antagonist ( P25101 ) with high selectivity for the endothelin-A ( ET(A) ) receptor to be approved for clinical use by regulatory agencies in Europe for the treatment of pulmonary arterial hypertension ( PAH ) . Clinical trials have shown it to be well tolerated and to improve exercise tolerance , functional class and pulmonary hemodynamics in PAH , results which appear to be at least as good as those for the mixed P25101 DB00559 . Importantly , compared to DB00559 , sitaxsentan has a lower incidence of liver toxicity and no interaction with sildenafil , a drug commonly used in the management of PAH . Furthermore , there is increasing evidence to suggest that ETRAs may play an important role in the future management of a wide variety of other conditions , from hypertension and renal disease to connective tissue disease and cancer . In some of these conditions , ET(A) selectivity may be an advantage . Inhibition of p53 transcriptional activity by the P04271 calcium-binding protein . The levels of S100 Ca(2+)-binding proteins correlate with the progression of certain tumors , but their role , if any , in carcinogenesis is still poorly understood . P04271 protein associates with both the p53 oligomerization domain ( residues 325-355 ) and the extreme C terminus of the tumor suppressor p53 ( residues 367-392 ) . Consequently , P04271 inhibits p53 tetramer formation and p53 phosphorylation mediated by protein kinase C , on p53 C-terminal end . In this report , we show that the P04271 protein decreases p53 DNA binding and transcriptional activity . The effect of P04271 is reflected in vivo by a reduced accumulation of p53 , P38936 , and Q00987 protein levels in co-transfection assays and in response to bleomycin . The P04271 can still interact with p53 in the absence of p53 extreme C-terminal end and reduce the expression of p53 downstream effector genes . These data indicate that P04271 does not require p53 extreme C-terminal end to inhibit p53 activity . Collectively , these findings imply that elevated levels of P04271 in tumors such as astrocytomas and gliomas could inhibit p53 functions and contribute to cancer progression . DB00855 dehydratase ( E.C. 4.2.1.24 ) : linkage analysis . Linkage data on aminolevulinate dehydratase ( P13716 , E.C. 4.2.1.24 ) and a series of other human genetic markers are presented . One hundred and two families ( 25 of them being informative ) from southwestern Germany were tested . Close linkage ( theta = 0.05 ) between P13716 and the following markers could be excluded : Rh , P36871 , Fy , P24666 , MNSs , HLA , Bf , GLO , O95394 , Jk , Pi , A6NDG6 , K , GPT . There is some evidence of possible linkage with Q9Y251 . Induced p21Cip1 in premature baboons with CLD : implications for alveolar hypoplasia . Aberrant pulmonary epithelial and mesenchymal cell proliferation occurs when newborns are treated with oxygen and ventilation to mitigate chronic lung disease . Because the cyclin-dependent kinase inhibitor P38936 inhibits proliferation of oxygen-exposed cells , its expression was investigated in premature baboons delivered at 125 days ( 67 % of term ) and treated with oxygen and ventilation pro re nata ( Q9H6Q4 ) for 2 , 6 , 14 , and 21 days . Approximately 5 % of all cells expressed P38936 during normal lung development of which < 1 % of these cells were pro-surfactant protein ( SP ) -B-positive epithelial cells . The percentage of cells expressing P38936 increased threefold in all Q9H6Q4 -treated animals , but different cell populations expressed it during disease progression . Between 2 and 6 days of treatment , P38936 was detected in 30-40 % of pro- P07988 cells . In contrast , only 12 % of pro- P07988 cells expressed P38936 by 14 and 21 days of treatment , by which time P38936 was also detected in mesenchymal cells . Even though increased epithelial and mesenchymal cell proliferation occurs during disease progression , those cells expressing P38936 did not also express the proliferative marker Ki67 . Thus two populations of epithelial and mesenchymal cells can be identified that are either expressing Ki67 and proliferating or expressing P38936 and not proliferating . These data suggest that P38936 may play a role in disorganized proliferation and alveolar hypoplasia seen in newborn chronic lung disease . Concurrent hypermethylation of multiple genes is associated with grade of oligodendroglial tumors . Current evidence suggests that epigenetic changes play an important role in the evolution of human cancers . In this study , we evaluated whether hypermethylation of CpG islands at the gene promotor regions of several tumor-related genes is involved in the carcinogenesis of oligodendroglial tumors . We examined the methylation status of 11 genes in a series of 43 oligodendroglial tumors ( 19 oligodendrogliomas , 13 anaplastic oligodendrogliomas , 9 oligoastrocytomas , and 2 anaplastic oligoastrocytomas ) by methylation-specific polymerase chain reaction . Our results showed that hypermethylation of CpG islands was detectable in 8 of 11 genes studied and 74 % of tumors were hypermethylated in at least 1 gene . Promotor hypermethylations were detected in O6-methylguanine-DNA methyltransferase ( P16455 ) , P06400 , estrogen receptor , p73 , p16INK4a , death-associated protein kinase , p15INK4b , and Q8N726 at 60 % , 34 % , 30 % , 16 % , 12 % , 10 % , 7 % , and 2 % , respectively . No hypermethylation was detected in the promotors of glutathione-S-transferase P1 , von Hippel-Lindau or the DNA mismatch repair ( hMLH1 ) genes . Statistical analysis revealed that concordant hypermethylation of at least 2 genes , p16INK4a and p15INK4b were significantly associated with anaplastic oligodendroglial tumors , and hypermethylation of P16455 was significantly associated with loss of chromosome 19q and with combined loss of chromosomes 1p and 19q . More importantly , several candidate tumor suppressor genes such as p16INK4a , p15INK4b , and p73 that were previously reported as unmutated in oligodendroglial tumors were found to be hypermethylated in their CpG islands . Taken together , we conclude that hypermethylation of CpG islands is a common epigenetic event that is associated with the development of oligodendroglial tumors . DB02546 and bortezomib synergistically cause ubiquitinated protein accumulation in prostate cancer cells . PURPOSE : Protein ubiquitination is a novel strategy used to treat malignancies . We investigated whether the histone deacetylase inhibitor vorinostat ( Cayman Chemical , Ann Arbor , Michigan ) and the proteasome inhibitor bortezomib ( LC Laboratories , Woburn , Massachusetts ) would synergistically cause the accumulation of ubiquitinated proteins in prostate cancer cells . MATERIALS AND METHODS : LNCaP , PC-3 and DU 145 cells ( ATCC™ ) were treated with vorinostat and/or bortezomib . Cell viability and induction of apoptosis were assessed . In vivo efficacy was evaluated in a murine subcutaneous tumor model using PC-3 cells . The influence of androgen receptor expression on bortezomib efficacy was examined using RNA interference . Changes in the expression of ubiquitinated proteins , cell cycle associated proteins and acetylated histone were evaluated . RESULTS : P10275 expression seemed to decrease bortezomib activity . PC-3 and DU 145 cells were more susceptible to bortezomib than LNCaP cells and the silencing of androgen receptor expression in LNCaP cells enhanced bortezomib activity . DB02546 and bortezomib synergistically induced apoptosis , inhibited prostate cancer cell growth and suppressed tumor growth in a murine xenograft model . The combination decreased cyclin D1 and cyclin-dependent kinase 4 expression , and increased P38936 expression . The combination synergistically caused the accumulation of ubiquitinated proteins and histone acetylation . This histone acetylation was a consequence of the accumulation of ubiquitinated proteins . CONCLUSIONS : DB02546 and bortezomib inhibit the growth of prostate cancer cells synergistically by causing ubiquitinated proteins to accumulate in cells . The current study provides a framework for testing the combination in patients with advanced prostate cancer . The ' cerebral diabetes ' paradigm for unipolar depression . Unipolar depression , alcoholism and suicide have become more common over the past decades . Genetic studies have attempted to link ( bipolar ) affective disorder to the short arm of chromosome 11 ( where the loci for insulin , insulin growth factor ( IGF ) , tyrosine hydroxylase ( TH ) and h-ras-oncogene are located ) but these have failed . Since TH and the insulin receptor require phosphorylation by protein kinases , then a defect of the h-ras-oncogene or its products ( P38936 ) could disorder both these systems and compromise catecholaminergic transmission in neurones and energy flow in glial cells . This could lead not only to a predisposition to depression ( ' trait markers ' ) but to neurotoxic damage , predisposed by inadequate cytosol Mg2+ levels of hypometabolism . Tyrosine , tryptophan and phenylalanine hydroxylases all require tetrahydrobiopterin ( BH4 ) which allosterically regulates its own activity as well as that of these enzymes . Anything which impairs this cofactor could lead to overt depression in predisposed individuals , and the heterocyclic amines are being increasingly implicated . These substances are derived from fried and broiled meats , azo food dyes , soft drinks and hard candies , but particularly from cigarette and petroleum fumes . The heterocyclic amines can inhibit aromatic-l-amino-acid-decarboxylase ( P20711 ) as well as the hydroxylases reversibly , but BH4 is inhibited noncompetitively . Thus , susceptible individuals ( those with inherited defective protein kinase phosphorylation ) might be ' tipped over ' by chronic exposure to these neurotoxins . The rising incidence of unipolar depression-associated morbidity could be significantly linked to increasing levels of heterocyclic amines in the developed nations . The solution structure of the bovine P04271 protein dimer in the calcium-free state . BACKGROUND : P04271 ( S100beta ) is a member of the S100 family of small calcium-binding proteins : members of this family contain two helix-loop-helix calcium-binding motifs and interact with a wide range of proteins involved mainly in the cytoskeleton and cell proliferation . P04271 is a neurite-extension factor and levels of P04271 are elevated in the brains of patients with Alzheimer 's disease or Down 's syndrome : the pattern of P04271 overexpression in Alzheimer 's disease correlates with the pattern of neuritic-plaque formation . Identification of a growing class of S100 proteins and the likely neurochemical importance of P04271 make the determination of the structure of P04271 of interest . RESULTS : We have used NMR to determine the structure of the reduced P04271 homodimer in the absence of calcium . Each monomer consists of a four-helix bundle , arranged in the dimer in an antiparallel fashion . The fourth helix of each monomer runs close to the equivalent helix of the other monomer for almost its full length , extending the hydrophobic core through the interface . The N-terminal , but not the C-terminal , calcium-binding loop is similar to the equivalent loop in the monomeric S100 protein calbindin and is in a conformation ready to bind calcium . CONCLUSIONS : The novel dimer structure reported previously for calcyclin ( P06703 ) is the common fold for the dimeric P04271 proteins . DB01373 binding to the C-terminal calcium-binding loop would be expected to require a conformational change , which might provide a signal for activation . The structure suggests regions of the molecule likely to be involved in interactions with effector molecules . Pharmacokinetic profiles of the novel P35354 selective inhibitor cimicoxib in dogs . DB05095 ( CX ) is a novel imidazole derivative that is a cyclo-oxygenase ( P36551 ) -2 selective non-steroidal anti-inflammatory drug and the latest P35354 selective inhibitor to be released for veterinary use . Currently there is limited information available on the pharmacokinetic ( PK ) properties of CX . The aim of the current study was to evaluate the PK features of CX after administration of the recommended dose and after administration of a more variable dose rate in the form of the commercially available tablet . In addition , the effects of food intake on the PK properties were also evaluated . In the first study , five healthy Beagle dogs received 2mg/kg CX via the oral route following a period of fasting . The second study was conducted using six healthy Labrador retriever dogs which each received an 80 mg tablet ( approximate dose 1.95-2.5mg/kg ) using a crossover design , both in the fasted and fed condition . The plasma concentrations of CX were detected by a validated HPLC method . No adverse effects were observed in any dogs during the experiment . The results from the PK analysis were similar between the studies , regardless of precision of dose and fasted and fed conditions . The mean peak concentration of CX was 0.49 and 0.43 μg/mL under fasted and fed conditions , respectively . The mean half-life was about 3h after all treatments . In addition , simulated multiple dosing data revealed that time over minimal effective concentration was similar after 1.95 , 2.0 and 2.5mg/kg dose administrations . These findings suggest that slight variation from the recommended dose should not alter the therapeutic outcome . In addition , CX can be administered to fed dogs without significantly affecting blood levels . The P38936 codon 31*C- and P14416 codon 313*T-related genotypes/alleles , but not P18887 codon 399 , hOGG1 codon 326 , and P21728 -48 polymorphisms , are correlated with the presence of leiomyoma . OBJECTIVE : To investigate whether the gene polymorphisms for P38936 , X-ray repair cross-complementing group 1 ( P18887 ) , human 8-oxoguanine glycosylase 1 ( hOGG1 ) , and dopamine D1 and D2 receptors ( P21728 , -2 ) are associated with leiomyoma susceptibility . DESIGN : Prospective study . SETTING : Departments of gynecology and genetics in a medical center . PATIENT(S) : Women were divided into two groups : leiomyoma ( n = 120 ) and nonleiomyoma ( n = 112 ) . INTERVENTION(S) : The P38936 codon 31 , P18887 codon 399 , hOGG1 codon 326 , P21728 -48 , and P14416 codon 313 polymorphisms were genotyped by polymerase chain reaction with restriction enzyme digestions ( Blp I , MspI , Fnu4HI , Dde I , and NcoI , respectively ) . MAIN OUTCOME MEASURE(S) : Genotypes and allelic frequencies . RESULT(S) : The P38936 codon 31(*)C- and P14416 codon 313(*)T-related genotypes/alleles were associated with the presence of leiomyomas . The proportions of P38936 (*)CC/CA/AA and P14416 (*)CC/CT/TT in both groups were 27.5/68.3/4.2 % and 12.5/51.7/35.8 % ( leiomyoma ) ; and 14.3/51.8/33.9 % and 33.9/40.2/25.9 % ( nonleiomyoma ) . P18887 , hOGG1 , and P21728 were not correlated with the presence of leiomyomas . P18887 (*)GG/GA/AA , hOGG1(*)TT/TA/AA , and P21728 (*)GG/GA/AA were 54.2/37.5/8.3 % , 36.7/44.2/19.1 % , and 3.3/25.8/70.8 % ( leiomyoma ) ; and 48.2/47.3/4.5 % , 43.6/41/15.4 % , and 3.6/25/71.4 % ( nonleiomyoma ) . CONCLUSION(S) : The P38936 codon 31(*)C- and P14416 codon 313(*)T-related genotypes/alleles were associated with the presence of leiomyoma . P18887 , hOGG1 , and P21728 were not correlated with leiomyoma development . Gene transcription abnormalities in canine atopic dermatitis and related human eosinophilic allergic diseases . Canine atopic dermatitis ( AD ) is clinically similar to human AD , implicating it as a useful model of human eosinophilic allergic disease . To identify cutaneous gene transcription changes in relatively early inflammation of canine AD , microarrays were used to monitor transcription in normal skin ( n=13 ) and in acute lesional AD ( P13716 ) and nearby visibly nonlesional AD ( NLAD ) skin ( n=13 ) from dogs . Scanning the putative abnormally transcribed genes , several potentially relevant genes , some abnormally transcribed in both NLAD and P13716 ( e.g. P05231 , Q8NET5 , Q9UJ68 , and P43405 ) , were observed . Comparison for abnormally transcribed genes common to two related human diseases , human AD and asthmatic chronic rhinosinusitis with nasal polyps ( aCRSwNP ) , further identified genes or gene sets likely relevant to eosinophilic allergic inflammation . These included : ( 1 ) genes associated with alternatively activated monocyte-derived cells , including members of the monocyte chemotactic protein ( MCP ) gene cluster , ( 2 ) members of the IL1 family gene cluster , ( 3 ) eosinophil-associated seven transmembrane receptor Q14246 and Q9BY15 genes , ( 4 ) interferon-inducible genes , and ( 5 ) keratin genes associated with hair and nail formation . Overall , numerous abnormally transcribed genes were observed only in canine AD ; however , many others are common to related human eosinophilic allergic diseases and represent therapeutic targets testable in dogs with AD . The role of tumor suppressor dysregulation in prostate cancer progression . P10275 activity is essential for prostate cancer development and progression . While there are classically defined roles for the retinoblastoma ( P06400 ) and p53 tumor suppressor pathways in maintenance of cell cycle control and the DNA damage response , recent studies have demonstrated a direct role of these two pathways in regulating AR expression and function . While the role of Pten deregulation in prostate cancer has provided much insight in to the mechanisms underlying prostate cancer initiation and progression , emerging roles for P06400 and p53 are likely to further expand upon our understanding of tumor suppressor/nuclear receptor interaction . As disconnecting mitogenic signaling from AR-mediated gene transcription underlies the progression to castrate resistant prostate cancer ( CRPC ) , functional inactivation of these two tumor suppressor pathways represents one mechanism through which AR protein levels can be upregulated and AR-mediated gene transcription can become aberrant . Importantly , recent advances in small molecule inhibitor design and discovery have led to the identification of agents capable of targeting these two prominent pathways and restoring the function of deregulated wild-type P06400 and p53 protein . While such agents have undergone extensive study in many solid tumor types , the additional importance of P06400 and p53 in restraining transcription of the AR gene within the prostate provides impetus for examining how loss of these two tumor suppressor proteins can facilitate transition of prostate cancers to CRPC . As will be reviewed in this article , restoration of P06400 and p53 functions are not only important in regard to shortterm cell cycle regulation and response to genomic stresses , but likely have direct implications for deregulation of the AR locus . Activation of serotonin receptor-2B rescues small-for-size liver graft failure in mice . The implantation of grafts below 30 % of the normal liver volume is associated with a high risk of failure known as small-for-size ( SFS ) syndrome . Strategies to rescue small grafts may have a dramatic impact on organ shortage . Serotonin is a potent growth factor for the liver . The goal of this study was to determine whether enhanced serotonin signaling could prevent the deleterious effects of SFS syndrome . We performed 30 % normal liver volume transplantations in wild-type C57/BL6 and interleukin-6 ( P05231 ) (-/-) mice . Some animals received α-methyl-5-HT ( DOI ) , an agonist of serotonin receptor-2 ( P41595 ) . Endpoints included long-term survival , serum and hepatic markers of liver injury and regeneration , assessment of hepatic microcirculation by intravital fluorescence microscopy and scanning electron microscopy , and transcript levels of a variety of serotonin receptors , tumor necrosis factor α , and P05231 . All recipients of small grafts ( controls ) died within 2-4 days of transplantation , whereas half of those receiving DOI survived permanently . Control animals disclosed major liver injury , including diffuse microvesicular steatosis in hepatocytes , impairment of microcirculation , and a failure of regeneration , whereas these parameters were dramatically improved in animals subjected to DOI . Blockage of P41595 blunted the protective effects of DOI . Whereas P05231 levels were higher in DOI-treated animals , P05231 (-/-) mice were still protected by DOI , suggesting a protective pathway independent of P05231 . CONCLUSION : Serotonin through its action on receptor-2B protects SFS liver grafts from injury and prevents microcirculation and regeneration . The mechanism of hepato-protection is independent of P05231 . P10275 -mediated regulation of the anti-atherogenic enzyme Q02318 involves the JNK/c-jun pathway . Q02318 , an enzyme with several important roles in cholesterol homeostasis and vitamin D₃ metabolism , has been ascribed anti-atherogenic properties . This study addresses an important problem regarding how this enzyme , involved in cholesterol metabolism in the liver and peripheral tissues , is regulated . Our results identify the human Q02318 gene as a new target for the JNK/c-jun pathway . Initial experiments showed that an inhibitor of c-Jun N-terminal kinase ( JNK ) downregulated basal Q02318 promoter activity whereas overexpression of JNK slightly enhanced promoter activity . P10275 ( AR ) -mediated upregulation of mRNA levels and endogenous enzyme activity was recently reported . In the present study , the AR antagonist nilutamide blocked the androgen induction of Q02318 . The present data revealed that inhibition of the JNK/c-jun pathway abolishes the AR-mediated effect on Q02318 transcription and enzyme activity , whereas overexpression of JNK markedly increased androgenic upregulation of Q02318 . In conclusion , the current results indicate involvement of the JNK/c-jun pathway in AR-mediated upregulation of human Q02318 . The link to JNK signaling is interesting since inflammatory processes may upregulate Q02318 to clear cholesterol from peripheral tissues . P20711 inhibition does not influence the diuretic and natriuretic response to exogenous alpha-atrial natriuretic peptide in man . The role of dopamine synthesis in the renal actions of human alpha-atrial natriuretic peptide ( alpha P01160 ) was investigated in six dehydrated volunteers using the P20711 inhibitor carbidopa . Each subject received oral placebo or carbidopa ( 100 mg ) followed by an infusion of alpha P01160 10 pmol.kg-1.min-1 for 1 h . The responses to placebo alone and to carbidopa alone were investigated on separate occasions. alpha P01160 produced a similar increase in plasma immunoreactive alpha P01160 whether placebo or carbidopa pretreatment had been given . Urinary dopamine excretion was increased by alpha P01160 . DB00190 pretreatment substantially attenuated this increase without affecting the natriuretic or water-diuretic response to alpha P01160 . DB00190 also failed to alter the change in filtration fraction produced by alpha P01160 . The results suggest that increased synthesis of intrarenal dopamine is not required for the renal effects of alpha P01160 in man . P12004 D1 : mechanism and consequence of androgen receptor co-repressor activity . P10275 regulation is pivotal for prostate growth and development . Activation of the receptor is dictated by association with androgen ( ligand ) and through interaction with co-activators and co-repressors . We have shown previously that cyclin D1 functions as a co-repressor to inhibit ligand-dependent androgen receptor activation . We demonstrate that cyclin D1 directly binds the N terminus of the androgen receptor and that this interaction is independent of ligand . Furthermore , we show that the interaction occurs in the nucleus and does not require the LXXLL motif of cyclin D1 . Although two distinct transactivation domains exist in the N terminus ( P38484 and AF-5 ) , the data shown support the hypothesis that cyclin D1 targets the P38484 transactivation function . The constitutively active AF-5 domain was refractory to cyclin D1 inhibition . By contrast , cyclin D1 completely abolished androgen receptor activity , even in the presence of potent androgen receptor co-activators . This action of cyclin D1 at least partially required de-acetylase activity . Finally , we show that transient , ectopic expression of cyclin D1 results in reduced cell cycle progression in androgen-dependent LNCaP cells independent of P11802 association . Collectively , our data support a model wherein cyclin D1 has a mitogenic ( P11802 -dependent ) function and an anti-mitogenic function ( dependent on regulation of the P38484 domain ) that can collectively control the rate of androgen-dependent cellular proliferation . These findings provide insight into the non-cell cycle functions of cyclin D1 and provide the impetus to study its pleiotropic effects in androgen-dependent cells , especially prostatic adenocarcinomas . A charybdotoxin-sensitive , Ca(2+)-activated K+ channel with inward rectifying properties in brain microvascular endothelial cells : properties and activation by endothelins . A charybdotoxin-sensitive , Ca(2+)-activated K+ channel was identified in cultured rat brain capillary endothelial cells by using conventional single-channel recording techniques and 86( P06400 +)-influx and efflux experiments . Channel activity was dependent on the presence of Ca2+ on the cytosolic face of the membrane with a threshold concentration of 100 nM . It was inhibited by charybdotoxin ( IC50 30 nM ) and quinine ( IC50 0.1 mM ) but not by apamin . K(Ca) channels showed unusual inward rectifying properties under asymmetrical ionic conditions . They were activated by endothelin-1 ( EC50 0.7 nM ) and endothelin-3 ( EC50 7-10 nM ) . The actions of endothelins were prevented by BQ-123 ( Ki = 8 nM ) in a competitive fashion , hence suggesting the involvement of an P25101 -receptor subtype . The channel activity was unaffected by cyclic AMP- or cyclic GMP-elevating agents . The possible role of the intermediate conductance , Ca(2+)-activated K+ channels for mediating K+ movements across the blood-brain barrier is discussed . An integrative in silico approach for discovering candidates for drug-targetable protein-protein interactions in interactome data . BACKGROUND : Protein-protein interactions ( PPIs ) are challenging but attractive targets for small chemical drugs . Whole PPIs , called the ' interactome ' , have been emerged in several organisms , including human , based on the recent development of high-throughput screening ( HTS ) technologies . Individual PPIs have been targeted by small drug-like chemicals ( SDCs ) , however , interactome data have not been fully utilized for exploring drug targets due to the lack of comprehensive methodology for utilizing these data . Here we propose an integrative in silico approach for discovering candidates for drug-targetable PPIs in interactome data . RESULTS : Our novel in silico screening system comprises three independent assessment procedures : i ) detection of protein domains responsible for PPIs , ii ) finding P18827 -binding pockets on protein surfaces , and iii ) evaluating similarities in the assignment of Gene Ontology ( GO ) terms between specific partner proteins . We discovered six candidates for drug-targetable PPIs by applying our in silico approach to original human PPI data composed of 770 binary interactions produced by our HTS yeast two-hybrid ( HTS-Y2H ) assays . Among them , we further examined two candidates , P19793 / P48552 and P24941 / P38936 , with respect to their biological roles , PPI network around each candidate , and tertiary structures of the interacting domains . CONCLUSION : An integrative in silico approach for discovering candidates for drug-targetable PPIs was applied to original human PPIs data . The system excludes false positive interactions and selects reliable PPIs as drug targets . Its effectiveness was demonstrated by the discovery of the six promising candidate target PPIs . Inhibition or stabilization of the two interactions may have potential therapeutic effects against human diseases . Role of the androgen receptor axis in prostate cancer . P10275 ( AR ) is expressed in nearly all prostate cancers , including treatment-refractory disease . The role of this receptor in the molecular endocrinology of prostate cancer has become increasingly clear in recent years . The AR is now known to participate in tumor progression through 3 mechanisms : expression ( activation and upregulation of receptor activity ) , point mutations , and ligand-independent activation . With regard to the latter mechanism , interleukin-6 ( P05231 ) is among the most important nonsteroidal regulators of AR activity . In the absence of androgen , P05231 causes activation of AR that is approximately 50 % of the maximal activity induced by androgen . At low concentrations of androgen , P05231 and androgen synergistically activate AR . Nonsteroidal antiandrogens usually antagonize this activation , but they switch to an agonist effect in the presence of oncostatin M , an P05231 -related cytokine . The growth of parental LNCaP cells is initially inhibited by exposure to P05231 , but long-term treatment renders the cells resistant to such inhibition and confers a growth advantage . Both P05231 and oncostatin M stimulate AR activity , but only oncostatin M is associated with strong acquisition of the agonist properties of nonsteroidal antiandrogens . It is hoped that continuing research on AR expression and function in prostate cancer will pave the way for new therapeutic strategies . [(11)C]5-HTP and microPET are not suitable for pharmacodynamic studies in the rodent brain . The PET tracer [(11)C]5-hydroxytryptophan ( [(11)C]5-HTP ) , which is converted to [(11)C]5-hydroxytryptamine ( [(11)C]5-HT ) by aromatic amino acid decarboxylase ( P20711 ) , is thought to measure 5-HT synthesis rates . But can we measure these synthesis rates by kinetic modeling of [(11)C]5-HTP in rat ? Male rats were scanned with [(11)C]5-HTP ( 60 minutes ) after different treatments . Scans included arterial blood sampling and metabolite analysis . 5-HT synthesis rates were calculated by a two-tissue compartment model ( 2TCM ) with irreversible tracer trapping or Patlak analysis . DB00190 ( inhibitor peripheral P20711 ) dose-dependently increased [(11)C]5-HTP brain uptake , but did not influence 2TCM parameters . Therefore , 10 mg/kg carbidopa was applied in all subsequent study groups . These groups included treatment with NSD 1015 ( general P20711 inhibitor ) or p-chlorophenylalanine ( PCPA , inhibitor of tryptophan hydroxylase , P17752 ) . In addition , the effect of a low-tryptophan ( DB00150 ) diet was investigated . NSD 1015 or DB00150 depletion did not affect any model parameters , but PCPA reduced [(11)C]5-HTP uptake , and the k3 . This was unexpected as NSD 1015 directly inhibits the enzyme converting [(11)C]5-HTP to [(11)C]5-HT , suggesting that trapping of radioactivity does not distinguish between parent tracer and its metabolites . As different results have been acquired in monkeys and humans , [(11)C]5-HTP-PET may be suitable for measuring 5-HT synthesis in primates , but not in rodents . A calcium sensor in the sodium channel modulates cardiac excitability . Sodium channels are principal molecular determinants responsible for myocardial conduction and maintenance of the cardiac rhythm . DB01373 ions ( Ca2+ ) have a fundamental role in the coupling of cardiac myocyte excitation and contraction , yet mechanisms whereby intracellular Ca2+ may directly modulate Na channel function have yet to be identified . Here we show that calmodulin ( P62158 ) , a ubiquitous Ca2+-sensing protein , binds to the carboxy-terminal ' IQ ' domain of the human cardiac Na channel ( hH1 ) in a Ca2+-dependent manner . This binding interaction significantly enhances slow inactivation-a channel-gating process linked to life-threatening idiopathic ventricular arrhythmias . Mutations targeted to the IQ domain disrupted P62158 binding and eliminated Ca2+/ P62158 -dependent slow inactivation , whereas the gating effects of Ca2+/ P62158 were restored by intracellular application of a peptide modelled after the IQ domain . A naturally occurring mutation ( A1924T ) in the IQ domain altered hH1 function in a manner characteristic of the Brugada arrhythmia syndrome , but at the same time inhibited slow inactivation induced by Ca2+/ P62158 , yielding a clinically benign ( arrhythmia free ) phenotype . In utero and lactational exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin alters postnatal development of seminal vesicle epithelium . 2,3,7,8-Tetrachlorodibenzo-p-dioxin ( TCDD ) has been shown to alter male reproductive development of laboratory animals through in utero and lactational exposure . As a result of exposure , the accessory glands of the male reproductive tract , including the seminal vesicle , are decreased in size as determined by total weight of the tissue . Analysis of seminal vesicle weights over time suggests that the changes may be transient . Administration of 1.0 microg/kg TCDD during gestation caused a significant decrease in seminal vesicle weights of offspring 8-11 months of age . We examined the effects of TCDD on seminal vesicles from rats exposed in utero and lactationally . Pregnant Long Evans rats were gavaged on gestation day 15 with 1.0 microg/kg TCDD in corn oil . Male pups were euthanized and necropsied on postnatal days ( P01160 ) 15 , 25 , 32 , 49 , 63 , and 120 . Seminal vesicles were weighed and then fixed in 10 % neutral buffered formalin and processed for microscopic examination . Seminal vesicle weights were not significantly decreased until P01160 32 . P10275 mRNA expression in P01160 25 seminal vesicles was not different from control . In the present study , TCDD exposure decreased seminal vesicle epithelial branching and differentiation . Control epithelial cells had tall columnar morphology with relatively abundant cytoplasm , whereas TCDD-treated cells had rounded nuclei and less cytoplasm . In addition , immunolocalization of proliferating nuclear antigen was confined to undifferentiated basal epithelial cells of controls but was found in both basal and luminal cells of the treated seminal vesicle . Results indicate that the TCDD-induced impaired growth of the rat seminal vesicles is associated with a dramatic decrease in the development of the epithelium . Murine cystathionine gamma-lyase : complete cDNA and genomic sequences , promoter activity , tissue distribution and developmental expression . P32929 ( CSE ) is the last key enzyme in the trans-sulphuration pathway for biosynthesis of cysteine from methionine . DB00151 could be provided through diet ; however , CSE has been shown to be important for the adequate supply of cysteine to synthesize glutathione , a major intracellular antioxidant . With a view to determining physiological roles of CSE in mice , we report the sequence of a complete mouse CSE cDNA along with its associated genomic structure , generation of specific polyclonal antibodies , and the tissue distribution and developmental expression patterns of CSE in mice . A 1.8 kb full-length cDNA containing an open reading frame of 1197 bp , which encodes a 43. P07988 , was isolated from adult mouse kidney . A 35 kb mouse genomic fragment was obtained by lambda genomic library screening . It contained promoter regions , 12 exons , ranging in size from 53 to 579 bp , spanning over 30 kb , and exon/intron boundaries that were conserved with rat and human CSE . The GC-rich core promoter contained canonical TATA and CAAT motifs , and several transcription factor-binding consensus sequences . The CSE transcript , protein and enzymic activity were detected in liver , kidney , and , at much lower levels , in small intestine and stomach of both rats and mice . In developing mouse liver and kidney , the expression levels of CSE protein and activity gradually increased with age until reaching their peak value at 3 weeks of age , following which the expression levels in liver remained constant , whereas those in kidney decreased significantly . Immunohistochemical analyses revealed predominant CSE expression in hepatocytes and kidney cortical tubuli . These results suggest important physiological roles for CSE in mice . Biological characterization of DB05037 , a small-molecule inhibitor of cyclin-dependent kinases , in human tumor cell lines . P12004 -dependent kinases ( CDK ) , and their regulatory cyclin partners , play a central role in eukaryotic cell growth , division , and death . This key role in cell cycle progression , as well as their deregulation in several human cancers , makes them attractive therapeutic targets in oncology . A series of CDK inhibitors was developed using Astex 's fragment-based medicinal chemistry approach , linked to high-throughput X-ray crystallography . A compound from this series , designated DB05037 , is currently in early-phase clinical development . We describe here the biological characterization of DB05037 , a potent inhibitor of several CDK family members . DB05037 showed potent antiproliferative activity ( 40-940 nmol/L ) in a panel of human tumor cell lines , and the mechanism of action was shown here to be consistent with the inhibition of P06493 and P24941 in solid tumor cell lines . DB05037 caused cell cycle arrest followed by apoptosis in human tumor cells and inhibited tumor growth in human tumor xenograft models . Tumor regression was observed following twice daily dosing of DB05037 in the HCT116 and HT29 colon cancer xenograft models . We show that these biological effects are linked to inhibition of CDKs in vivo and that DB05037 induces tumor cell apoptosis in these xenograft models . DB05037 has an attractive biological profile for development as a clinical candidate , and the tolerability and efficacy in animal models compare favorably with other CDK inhibitors in clinical development . Studies described here formed the biological rationale for investigating the potential therapeutic benefit of DB05037 in cancer patients . Immunohistochemical analysis of carcinomatous and sarcomatous components in the uterine carcinosarcoma : a case report . Uterine carcinosarcoma ( malignant mixed Mullerian tumor ) is an uncommon female genital tract neoplasm characterized by an admixture of epithelial and stromal malignant cells . We report a case of 50-year-old peri-menopausal woman diagnosed to have early-stage ( IB due to FIGO ) uterine carcinosarcoma of the homologous type with superficial ( 3mm ) myo-invasion . The patient showed no clinical symptoms of the disease and had no family history of female genital tract malignancies . Positive immunostaining for steroid receptors ( estrogen-alpha and progesterone receptors ) , cytokeratin , and P00533 was detected only in the carcinomatous area , whereas beta-catenin , BCL-2 , P35354 , p16(INK4a) , P60484 , Q8IUH3 , and vimentin were immunoreactive in both components . P10275 , CD10 , desmin , HER-2/neu , and P04637 were found to be negative either in the carcinomatous or in the sarcomatous area . Tumor proliferative activity was higher in the carcinomatous ( 25 % ) than in the sarcomatous ( 2 % ) component . Based on these findings , immunohistochemical evaluation of multiple receptor status in the carcinomatous and sarcomatous areas of carcinosarcoma may provide a clue to the pathogenesis and hormonal receptor status of this uncommon uterine malignancy . Altered transcriptional regulators in response to serum in immortalized lymphocytes from Alzheimer 's disease patients . Cell cycle disturbances may precede neuronal death in Alzheimer 's disease ( AD ) . We described alterations , in lymphocytes from AD patients , on the activity of two transcription factors , E2F and NF-kappaB , involved in cell proliferation and survival regulation , demonstrating that cell cycle dysfunction also occurs in peripheral cells . The analysis of E2F-DNA binding activity revealed lower signal intensity of protein-DNA complexes in AD cells , which correlated with increased phosphorylation of retinoblastoma ( P06400 ) related proteins and enhanced proliferation . The calmodulin ( P62158 ) antagonist calmidazolium ( DB01489 ) abrogated the increased activity of AD cells by partially dephosphorylating P06400 and Q08999 . The NF-kappaB-DNA binding activity increased as cell progress through the cell cycle . The reduced NF-kappaB activation observed in AD cells appears not to be related to the increased phosphorylation of the P06400 family proteins nor with the enhanced proliferative activity of AD cells , but seems to protect them from death induced by the loss of trophic support . Ca2+/ P62158 antagonists rescue NF-kappaB-DNA binding activity and sensitize AD cells to serum withdrawal . These observations suggest that disruption of Ca2+/ P62158 signaling pathway could be linked mechanistically to its pro cell survival actions , promoting enhanced proliferation or decreased cell death depending on the presence of growth-stimulatory signals . P10275 expressing neurons that project to the paraventricular nucleus of the hypothalamus in the male rat . Androgen receptors are distributed throughout the central nervous system and are contained by a variety of nuclei that are known to project to or regulate the paraventricular nucleus ( PVN ) of the hypothalamus , the final common pathway by which the brain regulates the hypothalamic-pituitary-adrenal ( Q9Y251 ) response to homeostatic threat . Here we characterized androgen receptor staining within cells identified as projecting to the PVN in male rats bearing iontophoretic or crystalline injections of the retrograde tracer FluoroGold aimed at the caudal two-thirds of the nucleus , where corticotropin-releasing hormone-expressing neurons are amassed . P10275 ( AR ) and FluoroGold ( FG ) double labeling was revealed throughout the limbic forebrain , including scattered numbers of cells within the anterior and posterior subdivisions of the bed nuclei of the stria terminalis ; the medial zone of the hypothalamus , including large numbers of AR-FG-positive cells within the anteroventral periventricular and medial preoptic cell groups . Strong and consistent colabeling was also revealed throughout the hindbrain , predominantly within the periaqueductal gray and the lateral parabrachial nucleus , and within various medullary cell groups identified as catecholaminergic , predominantly C1 and A1 neurons of the ventral medulla . These connectional data predict that androgens can act on a large assortment of multimodal inputs to the PVN , including those involved with the processing of various types of sensory and limbic information , and provide an anatomical framework for understanding how gonadal status could contribute to individual differences in Q9Y251 function . Identification of N-(4-piperidinyl)-4-(2,6-dichlorobenzoylamino)-1H-pyrazole-3-carboxamide ( DB05037 ) , a novel cyclin dependent kinase inhibitor using fragment-based X-ray crystallography and structure based drug design . The application of fragment-based screening techniques to cyclin dependent kinase 2 ( P24941 ) identified multiple ( > 30 ) efficient , synthetically tractable small molecule hits for further optimization . Structure-based design approaches led to the identification of multiple lead series , which retained the key interactions of the initial binding fragments and additionally explored other areas of the DB00171 binding site . The majority of this paper details the structure-guided optimization of indazole ( 6 ) using information gained from multiple ligand- P24941 cocrystal structures . Identification of key binding features for this class of compounds resulted in a series of molecules with low nM affinity for P24941 . Optimisation of cellular activity and characterization of pharmacokinetic properties led to the identification of 33 ( DB05037 ) , which is currently being evaluated in clinical trials for the treatment of human cancers . The mechanism of the G0/ P55008 cell cycle phase arrest induced by activation of O75469 in human cells . CONTEXT : O75469 ( O75469 ) is an important transcriptional regulator that plays important roles in the cell metabolism and cell growth by regulating the transcriptional of a sort of metabolizing enzymes . OBJECTIVE : To investigate whether rifampicin effected HepG2 cells growth and the inhibition was due to the G0/ P55008 phase arrest . METHODS : O75469 -knockdown experiments using RNAi showed that the cell cycle phase arrest mediated by rifampicin based on activation of O75469 . The results also indicated that cell phase arrest by rifampicin could protect cells form UVB-induced DNA damage . P19793 ( RXRα ) expression level in cells is another key factor for cell cycle phase arrest mediated by rifampicin . Both over expression and lacking expression of RXRα in cell reduced the cell arrest efficiency mediated by rifampicin . In the study , we found that rifampicin inhibited HepG2 cells growth and demonstrated that the inhibition is due to the G0/ P55008 phase arrest through flow cytometry analysis . CONCLUSION : The results showed that RXRα promote cell cycle phase transition rate of HepG2 . Competitive bind of rifampicin-activated O75469 with RXRα is one main reason to arrest cell cycle phase through inhibiting combination of RXRα with other partners . DB01045 could promote cell growth rate when RXRα expressed more excessively than O75469 in cells . Endothelin receptor antagonists as disease modifiers in systemic sclerosis . Systemic sclerosis ( SSc ) is a multisystem connective tissue disease of unknown etiology that is characterized by inflammation , vascular dysfunction and fibrosis of the skin and visceral organs . SSc is clinically diverse both in terms of the burden of skin and organ involvement and the rate of progression of the disease . Recent studies indicate that the endothelin system , especially ET-1 and the P25101 and ETB receptors may play a key role in the pathogenesis of SSc . A new class of drugs , endothelin receptor antagonists has been introduced for treatment of patients with pulmonary arterial hypertension ( PAH ) . DB00559 , a dual endothelin receptor antagonist as well as DB06268 and DB06403 , selective blockers of the P25101 receptor have proven effective in SSc-PAH . This effect may be mediated through both a vasodilatory and antifibrotic effect , thus making these agents attractive as potential disease modifying agents for SSc . P10275 rediscovered : the new biology and targeting the androgen receptor therapeutically . Discoveries over the past decade suggest that castration-resistant prostate cancer ( CRPC ) is sensitive , but not resistant to , further manipulation of the androgen-androgen receptor ( AR ) axis . Several new therapies that target this axis have demonstrated clinical activity . In this article , preclinical and clinical findings occurring in the field of AR-targeted therapies are reviewed . Reviews of scientific and clinical development are divided into those occurring prereceptor ( androgen production and conversion ) and at the level of the receptor ( AR aberrations and therapies targeting AR directly ) . Intracrine androgen production and AR amplification , among others , are among the principal aberrancies driving CRPC growth . Phase III data with abiraterone acetate and phase II data with DB08899 , along with other similar therapies , confirm for the clinician that the scientific findings related to persistent AR signaling in a castrate milieu can be harnessed to produce significant clinical benefit for patients with the disease . Studies aimed at optimizing the timing of their use and exploring the mechanisms of resistance to these therapies are under way . The clinical success of therapies that directly target androgen synthesis as well as the most common aberrancies of the AR confirm that prostate cancer retains dependence on AR signaling , even in the castrate state . Lipoic acid inhibits the DNA repair protein O 6-methylguanine-DNA methyltransferase ( P16455 ) and triggers its depletion in colorectal cancer cells with concomitant autophagy induction . Alkylating agents are present in food and tobacco smoke , but are also used in cancer chemotherapy , inducing the DNA lesion O (6)-methylguanine . This critical adduct is repaired by O (6)-methylguanine-DNA methyltransferase ( P16455 ) , resulting in P16455 inactivation and degradation . In the present study , we analyzed the effects of the natural disulfide compound lipoic acid ( LA ) on P16455 in vitro and in colorectal cancer cells . We show that LA , but not its reduced form dihydrolipoic acid , potently inhibits the activity of recombinant P16455 by interfering with its catalytic DB00151 -145 residue , which was partially reversible by N-acetyl cysteine . Incubation of HCT116 colorectal cancer cells with LA altered their glutathione pool and caused a decline in P16455 activity . This was mirrored by LA-induced depletion of P16455 protein , which was not attributable to changes in P16455 messenger RNA levels . Loss of P16455 protein coincided with LA-induced autophagy , a process resulting in lysosomal degradation of proteins , including presumably P16455 . LA-stimulated autophagy in a p53-independent manner as revealed by the response of isogenic HCT116 cell lines . Knockdown of the crucial autophagy component beclin-1 and chemical inhibitors blocked LA-induced autophagy , but did not abrogate LA-triggered P16455 degradation . Concomitant with P16455 depletion , LA pretreatment resulted in enhanced O (6)-methylguanine levels in DNA . It also increased the cytotoxicity of the alkylating anticancer drug temozolomide in temozolomide-resistant colorectal cancer cells . Taken together , our study showed that the natural compound LA inhibits P16455 and induces autophagy . Furthermore , LA enhanced the cytotoxic effects of temozolomide , which makes it a candidate for a supplement in cancer therapy . The centrality of P08397 expression levels on DB00855 efficacy . Successful 5-aminolevulinic acid-based photodynamic therapy ( DB00855 ) is dependent on efficient porphyrin synthesis in the inflicted cancer tissue , which is regulated by several enzymes . Irradiation of the tumor excites the light-sensitive porphyrins and results in ROS production and cell death . In this study we investigated the effect of the expression levels of two main enzymes in heme biosynthesis , ALA dehydratase ( P13716 ) and porphobilinogen deaminase ( P08397 ) , on the capacity of K562 cells to undergo cell death following DB00855 . We manipulated P08397 and P13716 expression levels by shRNAs and P08397 overexpressing plasmid . P08397 down-regulation induced an elevation in P13716 activity , while overexpression of P08397 reduced P13716 activity , indicating a novel regulation feedback of P08397 on P13716 activity . This feedback mechanism enabled partial PpIX synthesis under P08397 silencing , whereas P13716 silencing reduced PpIX production to a minimum . DB00855 efficacy was directly correlated to PpIX levels . Thus , only P13716 -silenced cells were not affected by ALA+ irradiation , while following P08397 silencing , the accumulated PpIX , though decreased , was sufficient for successful DB00855 . The alterations in P13716 activity level initiated by changes in P08397 expression indicates P08397 's central role in heme synthesis . This enables efficient DB00855 , even when P08397 is not fully active . Conversely , P13716 loss resulted in reduced PpIX synthesis and consequently failure in DB00855 , due to the absence of compensation mechanism for P13716 . NO-donor P35354 inhibitors . New nitrooxy-substituted 1,5-diarylimidazoles endowed with P35354 inhibitory and vasodilator properties . A series of NO-donor diarylimidazoles derived from the lead compound DB05095 were synthesized and evaluated for their P35354 inhibitory activity and their stability in whole blood as well as for vasodilator properties . The products are partly transformed into the corresponding alcohols following 24-h incubation in whole blood . All of them display good P23219 / P35354 selectivity , but are less potent than the lead ; a molecular modeling study was carried out to investigate their binding mode . The compounds are also capable of relaxing rat aorta strips precontracted with phenylephrine with a NO-dependent mechanism ; this property could confer reduced cardiotoxicity with respect to traditional P35354 inhibitors .

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Role of xanthine oxidoreductase and NAD(P)H oxidase in endothelial superoxide production in response to oscillatory shear stress . Oscillatory shear stress occurs at sites of the circulation that are vulnerable to atherosclerosis . Because oxidative stress contributes to atherosclerosis , we sought to determine whether oscillatory shear stress increases endothelial production of reactive oxygen species and to define the enzymes responsible for this phenomenon . Bovine aortic endothelial cells were exposed to static , laminar ( 15 dyn/cm2 ) , and oscillatory shear stress ( +/-15 dyn/cm2 ) . Oscillatory shear increased superoxide ( O2.- ) production by more than threefold over static and laminar conditions as detected using electron spin resonance ( P03372 ) . This increase in O2*- was inhibited by oxypurinol and culture of endothelial cells with tungsten but not by inhibitors of other enzymatic sources . DB05262 also prevented H2O2 production in response to oscillatory shear stress as measured by dichlorofluorescin diacetate and Amplex Red fluorescence . DB02134 -dependent O2*- production was increased in homogenates of endothelial cells exposed to oscillatory shear stress . This was associated with decreased xanthine dehydrogenase ( P47989 ) protein levels and enzymatic activity resulting in an elevated ratio of xanthine oxidase ( XO ) to P47989 . We also studied endothelial cells lacking the p47phox subunit of the NAD(P)H oxidase . These cells exhibited dramatically depressed O2*- production and had minimal XO protein and activity . Transfection of these cells with p47phox restored XO protein levels . Finally , in bovine aortic endothelial cells , prolonged inhibition of the NAD(P)H oxidase with apocynin decreased XO protein levels and prevented endothelial cell stimulation of O2*- production in response to oscillatory shear stress . These data suggest that the NAD(P)H oxidase maintains endothelial cell XO levels and that XO is responsible for increased reactive oxygen species production in response to oscillatory shear stress . DB02134 dehydrogenase and xanthine oxidase activity and gene expression in renal epithelial cells . Cytokine and steroid regulation . Reactive oxygen species have been implicated in the tissue injury and loss of epithelial barrier function associated with a number of clinical disorders in which disregulated inflammation seems to be a dominant event , such as endotoxemia and viral syndromes . In these disorders , xanthine oxidase ( XO ) contained within the epithelial cell has been proposed as a major source of injurious reactive oxygen species . This study was undertaken in an effort to understand the regulation of xanthine dehydrogenase ( P47989 ) /XO expression at both the activity and gene expression levels in the epithelial cell under conditions associated with the inflammatory response . The results indicate that P01375 , P01579 , P05231 , IL-1 , and dexamethasone induce P47989 /XO activity in bovine renal epithelial cells ( MDBK ) . This pattern of P47989 /XO regulation by cytokines and steroids is analogous to the profile of response seen by acute phase reactants . Metabolic labeling and immunoprecipitation revealed the increase in P47989 /XO activity requires new protein synthesis . By Northern analysis , all cytokines and dexamethasone increased the level of the 5-kb P47989 /XO mRNA . This increase was not detectable in the presence of actinomycin D but was further induced in the presence of cycloheximide , consistent with the major site of P47989 /XO up-regulation occurring at the transcriptional level . P47989 /XO mRNA was very stable , with no indication that the rates of transcript degradation contributed to differences in mRNA accumulation or ultimate activity levels . In addition to providing information on the regulation of P47989 /XO , the data presented furthers the understanding of the epithelial cell 's potential to actively respond to immunomodulators associated with injury/inflammation . The anti-androgen drug dutasteride renders triple negative breast cancer cells more sensitive to chemotherapy via inhibition of HIF-1α-/ P15692 -signaling . BACKGROUND : Triple negative breast cancer ( TNBC ) is characterized by lack of expression of both estrogen and progesterone receptor as well as lack of amplification of P04626 . Patients with TNBC carry an unfavorable prognosis compared to other breast cancer subtypes given that endocrine or P04626 targeted therapies are not effective , rendering chemotherapy the sole effective treatment option to date . Therefore , there is a high demand for additional novel treatment options . FINDINGS : We previously published a list of genes showing both higher gene expression rates in TNBC and , in addition , are known to encode targets of non-oncologic drugs . P18405 , which encodes the type-1 isoform of the steroid-5alpha-reductase , which is involved in androgen metabolism , was found to be one of these genes . DB01126 is a dual blocker of both the type-1 and type-2 isoform of P18405 and is indicated in the treatment of benign prostate hyperplasia . Treatment of TNBC cell lines with dutasteride was associated with a dose-dependent decrease in cell viability , altered protein expression of P15692 and HIF-1α and increased chemosensitivity . CONCLUSION : Our results demonstrate that the P18405 -corresponding anti-androgenic drug dutasteride might act as a combinatorial therapeutic option besides standard chemotherapy in highly aggressive TNBC . Beneficial effects of omega-3 fatty acids on the consequences of a fructose diet are not mediated by Q07869 delta or PGC1 alpha . PURPOSE : To study , in high-fructose-fed rats , the effect of a dietary enrichment in omega-3 polyunsaturated fatty acids ( n-3 PUFA ) on the expression of genes involved in lipid metabolism and cardiovascular function . METHODS : Twenty-eight male " Wistar Han " rats received for 8 weeks , either a standard chow food or an isocaloric 67 % fructose diet enriched or not in alpha-linolenic acid ( ALA ) or in docosahexaenoic ( DB01708 ) and eicosapentaenoic acids ( EPA ) mix ( DB01708 /EPA ) . After sacrifice , blood was withdrawn for biochemical analyses ; heart , periepididymal adipose tissue and liver were collected and analyzed for the expression of 22 genes by real-time PCR . RESULTS : DB04173 intake resulted in an increase in liver weight and triglyceride content , plasma triglyceride and cholesterol concentrations , although no difference in glucose and insulin . In the liver , lipogenesis was promoted as illustrated by an increase in stearoyl- DB01992 desaturase and fatty acid synthase ( Fasn ) together with a decrease in Q07869 gamma , delta and Q07869 gamma coactivator 1 alpha ( PGC1 alpha ) expression . In the heart , Fasn and Q07869 delta expression were increased . The addition of ALA or DB01708 /EPA into the diet resulted in a protection against fructose effects except for the decreased expression of PPARs in the liver that was not counterbalanced by n-3 PUFA suggesting that n-3 PUFA and fructose act independently on the expression of PPARs and PGC1 alpha . CONCLUSIONS : In liver , but not in heart , the fructose-enriched diet induces an early tissue-specific reduction in Q07869 gamma and delta expression , which is insensitive to n-3 PUFA intake and dissociated from lipogenesis . Pharmacokinetics and metabolism of a P35354 inhibitor , valdecoxib , in mice . The pharmacokinetics and metabolism of valdecoxib , a potent cyclooxygenase-2 selective inhibitor , were investigated in mice . DB00580 was extensively metabolized after a single 5 mg/kg oral administration of [(14)C]valdecoxib and elimination of unchanged drug was minor ( less than 1 % ) in male and female mice . The total mean percentage of administered radioactive dose recovered was 99.8 % in the male mice and 94.7 % in the female mice . Sixteen metabolites were identified in mouse plasma , red blood cells , urine , and feces . The main phase I metabolic pathway of valdecoxib in mice involved the oxidation of the 5-methyl group to form the active hydroxymethyl metabolite M1 . M1 was further oxidized to the carboxylic acid metabolite M4 , which underwent opening of the isoxazole ring to form M6 and M13 . Phase II metabolism included glucuronide , glucoside , and methyl sulfone conjugations . M1 was also conjugated with glucuronic acid and glucose to yield M-G and M1-glucose , respectively . Three novel methylsulfone conjugates M20 , M21 , and M21-G were detected in blood or urine . DB00580 and M1 were the major radioactive components in plasma and red blood cells . The plasma area under the curve from zero to infinity ( AUC(0-infinity) ) values for valdecoxib and M1 were 3.58 and 0.850 microg . h/ml in males and 2.08 and 1.63 microg . h/ml in females , respectively . The RBC AUC(0-infinity) values for valdecoxib and M1 were 12.1 and 22.6 microg . h/g in males and 6.42 and 35.2 microg . h/g in females , respectively . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . Differential increases in catecholamine metabolizing enzymes in amyotrophic lateral sclerosis . The activity of three catecholamine-metabolizing enzymes , monoamine oxidase type A and type B ( P21397 and P27338 ) as well as catechol-O-methyltransferase ( P21964 ) , were estimated in homogenates of human spinal cord using radiometric assays . The enzyme activities were determined in postmortem spinal cord tissue from controls and cases with amyotrophic lateral sclerosis ( P35858 ) . The activity of P21397 was below the limit of detectability in both controls and P35858 cases . The activities of P27338 and P21964 were evenly distributed at the various spinal levels . The P27338 activity was substantially elevated in P35858 spinal homogenates , whereas only a slight , but not statistically significant , increase in P21964 activity was observed . A significant correlation between P21964 and P27338 activities was observed for controls . However , this covariation was not apparent for the P35858 cases . These results suggest that the two enzyme proteins are regulated by more complex mechanisms in the spinal cord in amyotrophic lateral sclerosis than simple general increases caused by elevated astroglial cell numbers . In addition , the P21397 , P27338 , and P21964 activities were estimated in spinal cords from rats treated with the selective P27338 inhibitor L-deprenyl , a drug with putative neuroprotective effects in neurodegenerative disorders . After 3 weeks of L-deprenyl treatment ( 0.25 mg/kg/day , sc ) , the spinal P21397 and P27338 activities were decreased by 50 and 80 % , respectively . In contrast , the P21964 activity was not altered by L-deprenyl administration . Synthesis of new thiazolo[4,5-d]pyrimidines as DB01285 releasing factor modulators . P06850 ( CRF ) is a neurohormone that plays a crucial role in integrating the body 's overall response to stress . It appears necessary and sufficient for the organism to mount functional , physiological and endocrine responses to stressors . CRF is released in response to various triggers such as chronic stress . The role of CRF and its involvement in these neurological disorders suggest that new drugs that can target the CRF function or bind to its receptors may represent a new development of neuropsychiatric medicines to treat various stress-related disorders including depression , anxiety and addictive disorders . Based on pharmacophore of the CRF1 receptor antagonists , a new series of thiazolo[4,5-d] pyrimidines were synthesized as P06850 ( CRF ) receptor modulators and the prepared compounds carry groups shown to produce optimum binding affinity to CRF receptors . Twenty two compounds were evaluated for their CRF1 receptor binding affinity in P29320 293 cell lines and two compounds 5o and 5s showed approximately 25 % binding affinity to CRF1 receptors . Selected compounds ( 5c and 5f ) were also evaluated for their effect on expression of genes associated with depression and anxiety disorders such as CRF1 , P16220 , P21397 , P31645 , P01303 , DatSLC6a3 , and P09172 and significant upregulation of CRF1 mRNA has been observed with compound 5c . High-resolution X-ray structure of isoaspartyl dipeptidase from Escherichia coli . Q7L266 from Escherichia coli functions in protein degradation by catalyzing the hydrolysis of beta-L-isoaspartyl linkages in dipeptides . The best substrate for the enzyme reported thus far is iso- DB00128 - DB00149 . Here we report the X-ray analysis of the enzyme in its resting state and complexed with aspartate to 1.65 and 2.1 A resolution , respectively . The quaternary structure of the enzyme is octameric and can be aptly described as a tetramer of dimers . Each subunit folds into two distinct domains : the N-terminal region containing eight strands of mixed beta-sheet and the C-terminal motif that is dominated by a ( beta,alpha ) (8)-barrel . A binuclear zinc center is located in each subunit at the C-terminal end of the ( beta,alpha ) (8)-barrel . Ligands to the binuclear metal center include DB00117 68 , DB00117 70 , DB00117 201 , DB00117 230 , and DB00128 285 . The two zincs are bridged by a carboxylated lysine residue ( Lys 162 ) and a solvent molecule , most likely a hydroxide ion . The product of the reaction , aspartate , binds to the enzyme by displacing the bridging solvent with its side chain functional group . From this investigation it is proposed that the reaction mechanism of the enzyme proceeds through a tetrahedral intermediate and that the bridging solvent attacks the re face of the carbonyl carbon of the scissile peptide bond . This structural analysis confirms the placement of isoaspartyl dipeptidase into the urease-related amidohydrolase superfamily . Activity and expression of glutathione S-transferase pi in patients with amyotrophic lateral sclerosis . O60760 ( Q86UG4 , EC 2.5.1.18 ) is an enzyme responsible for inactivation of a large variety of toxic , electrophilic compounds and organic peroxides . Q86UG4 activity and Q86UG4 pi expression were studied in patients with amyotrophic lateral sclerosis ( P35858 ) . Studies were conducted on cerebrospinal fluid ( P04141 ) , blood serum and peripheral blood mononuclear cells ( PBMC ) obtained from 40 P35858 patients . P04141 from 30 subjects without neurological diseases and blood from 40 healthy blood donors were used as controls . Q86UG4 activity assayed with 1-chloro-2,4-dinitrobenzene ( substrate for transferase activity ) and cumene peroxide ( substrate for peroxidase activity ) was significantly decreased in PBMC of P35858 patients , as well as the Q86UG4 pi expression on both mRNA and protein level . The mean peroxidase activity was however significantly increased in P04141 and serum of P35858 patients with the specificity of 80 % and 73 % , and the sensitivity of 78 % and 85 % , respectively . It can thus be concluded that the protective barrier formed by Q86UG4 is originally affected in peripheral blood of P35858 patients , and may increase their vulnerability to toxic effects of electrophilic compounds and organic peroxides . Studies on a larger group are needed to answer the question whether DB00143 -Px determination may be implicated in the diagnosis of P35858 . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Novel positron emission tomography tracer distinguishes normal from cancerous cells . Development of tumor-specific probes for imaging by positron emission tomography has broad implications in clinical oncology , such as diagnosis , staging , and monitoring therapeutic responses in patients , as well as in biomedical research . P04818 ( P04818 ) -based de novo biosynthesis of DNA is an important target for drug development . Increased DNA replication in proliferating cancerous cells requires P04818 activity , which catalyzes the reductive methylation of DB03800 to dTMP using ( R ) -N(5),N(10)-methylene- DB00116 ( MTHF ) as a cofactor . In principle , radiolabeled MTHF can be used as a substrate for this reaction to identify rapidly dividing cells . In this proof-of-principle study , actively growing ( log phase ) breast cancer ( MCF7 , MDA-MB-231 , and hTERT- P31947 ) , normal breast ( human mammary epithelial and MCF10A ) , colon cancer ( HT-29 ) , and normal colon ( FHC ) cells were incubated with [(14)C]MTHF in culture medium from 30 min to 2 h , and uptake of radiotracer was measured . Cancerous cell lines incorporated significantly more radioactivity than their normal counterparts . The uptake of radioactively labeled MTHF depended upon a combination of cell doubling time , folate receptor status , S phase percentage , and P04818 expression in the cells . These findings suggest that the recently synthesized [(11)C]MTHF may serve as a new positron emission tomography tracer for cancer imaging . Genotype frequencies of 50 polymorphisms for 241 Japanese non-cancer patients . This paper lists the genotype frequencies of 50 polymorphisms of 37 genes ( P05091 , P07550 , P13945 , P21964 , P16671 , P25025 , P24385 , P35354 , P11509 , P05093 , P11511 , IGF1 , IL-1A , IL-1B , IL-1RN , IL-1R1 , P05231 , P10145 , P22301 , P41159 , Le , L-myc , P05164 , Q99707 , P42898 , P21397 , P15559 , O15527 , p53 , p73 , Se , P31213 , TGF-B , P01375 -A , P01375 -B , P18074 , and P18887 ) and 6 sets of combined genotype frequencies for 241 non-cancer Japanese outpatients . Though the genotype frequencies of 25 polymorphisms have already been reported in our previous papers , 15 polymorphisms ( P16671 A52C , P25025 C785T , P24385 G870A , IGF1 C/T at intron 2 and G2502T , IL-1A 46-bp VNTR , IL-1R1 C-116T , P05231 Ins/Del 17C , P10145 A-278T and C74T , IL- 10 T-819C , P41159 A-2548G , P31213 2-bp VNTR , P18074 Lys751Gln , and P18887 Arg399Gln ) and six sets of combined genotype frequencies ( IL-1B C-31T and IL-1A C-889T , IL-1B C-31T and IL-1RN 86-bp VNTR , IL-1B C-31T and IL-1R1 C-116T , P01375 -A G-308A and P01375 -B A252G , P31213 Val89Leu and 2-bp VNTR , and P18887 Arg399Gln and P18074 Lys751Gln ) were reported in this paper for the first time for Japanese . Although microarray technology will produce this kind of information in near future , this is the first document that reports the genotype/allele frequencies among Japanese for an archival purpose . Enhanced invasiveness of breast cancer cell lines upon co-cultivation with macrophages is due to P01375 dependent up-regulation of matrix metalloproteases . Apart from the neoplastic cells , malignant tumours consist of the extracellular matrix ( Q13201 ) and normal cells , in particular tumour-associated macrophages ( TAM ) . To understand the mechanisms by which TAM can influence tumour cell invasion we co-cultured the human breast cancer cell lines MCF-7 , SK-BR-3 and the benign mammary epithelial cell line hTERT- P31947 with macrophages . Co-incubation enhanced invasiveness of the tumour cells , while hTERT- P31947 remained non-invasive . Addition of the broad-spectrum matrix metalloprotease ( MMP ) -inhibitor FN 439 , neutralizing P14780 or tumour necrosis factor-alpha ( P01375 ) antibodies reduced invasiveness to basal levels . As shown by zymography , all cell lines produced low amounts of P08253 , -3 , -7 and -9 under control conditions . Basal MMP production by macrophages was significantly higher . Upon co-incubation , supernatant levels of MMPs -2 , -3 , -7 and -9 increased significantly , paralleled by an increase of P08253 activation . P08253 and -9 induction could be blocked by P01375 antibodies . Co-culture of macrophages and hTERT- P31947 did not lead to MMP induction . In the co-cultures , mRNAs for MMPs and P01375 were significantly up-regulated in macrophages , while the mRNA concentrations in the tumour cells remained unchanged . In summary , we have found that co-cultivation of tumour cells with macrophages leads to enhanced invasiveness of the malignant cells due to P01375 dependent MMP induction in the macrophages . DB00193 and another atypical opioid meperidine have exaggerated serotonin syndrome behavioural effects , but decreased analgesic effects , in genetically deficient serotonin transporter ( P31645 ) mice . The serotonin syndrome is a potential side-effect of serotonin-enhancing drugs , including antidepressants such as selective serotonin reuptake inhibitors ( SSRIs ) and monoamine oxidase inhibitors ( MAOIs ) . We recently reported a genetic mouse model for the serotonin syndrome , as serotonin transporter ( P31645 ) -deficient mice have exaggerated serotonin syndrome behavioural responses to the MAOI tranylcypromine and the serotonin precursor 5-hydroxy-l-tryptophan ( 5-HTP ) . As numerous case reports implicate the atypical opioids tramadol and meperidine in the development of the human serotonin syndrome , we examined tramadol and meperidine as possible causative drugs in the rodent model of the serotonin syndrome in P31645 wild-type ( +/+ ) , heterozygous ( +/- ) and knockout ( -/- ) mice . Comparisons were made with P31645 mice treated with either vehicle or morphine , an opioid not implicated in the serotonin syndrome in humans . Here we show that tramadol and meperidine , but not morphine , induce serotonin syndrome-like behaviours in mice , and we show that this response is exaggerated in mice lacking one or two copies of P31645 . The exaggerated response to tramadol in P31645 -/- mice was blocked by pretreatment with the P08908 antagonist WAY 100635 . Further , we show that morphine- , meperidine- and tramadol-induced analgesia is markedly decreased in P31645 -/- mice . These studies suggest that caution seems warranted in prescribing or not warning patients receiving SSRIs or MAOIs that dangerous side-effects may occur during concurrent use of tramadol and similar agents . These findings suggest that it is conceivable that there might be increased vulnerability in individuals with P31645 polymorphisms that may reduce P31645 by more than 50 % , the level in P31645 +/- mice . Q00613 inhibits expression of P05231 through activating transcription factor 3 . The febrile response is a complex physiological reaction to disease , including a cytokine-mediated increase in body temperature and the activation of inflammatory systems . Fever has beneficial roles in terms of disease prognosis , partly by suppressing the expression of inflammatory cytokines . However , the molecular mechanisms underlining the fever-mediated suppression of inflammatory gene expression have not been clarified . In this study , we showed that heat shock suppresses LPS-induced expression of P05231 , a major pyrogenic cytokine , in mouse embryonic fibroblasts and macrophages . Q00613 ( Q00613 ) activated by heat shock induced the expression of activating transcription factor ( P39905 ) 3 , a negative regulator of P05231 , and P18847 was necessary for heat-mediated suppression of P05231 , indicating a fever-mediated feedback loop consisting of Q00613 and P18847 . A comprehensive analysis of inflammatory gene expression revealed that heat pretreatment suppresses LPS-induced expression of most genes ( 86 % ) , in part ( 67 % ) via P18847 . When Q00613 -null and P18847 -null mice were injected with LPS , they expressed much higher levels of P05231 than wild-type mice , resulting in an exaggerated febrile response . These results demonstrate a novel inhibitory pathway for inflammatory cytokines . DB03758 activates heat shock protein expression and cardioprotection in neonatal rat cardiomyocytes . Heat shock proteins ( HSPs ) constitute an endogenous cellular defense mechanism against environmental stresses . In the past few years , studies have shown that overexpression of HSPs can protect cardiac myocytes against ischemia-reperfusion injury . In an attempt to increase the HSPs in cardiac tissue , we used the compound radicicol that activates HSP expression by binding to the P08238 kDa ( HSP90 ) . HSP90 is the main component of the cytosolic molecular chaperone complex , which has been implicated in the regulation of the heat shock factor 1 ( Q00613 ) . Q00613 is responsible for the transcriptional activation of the heat shock genes . In the present study , we show that radicicol induces HSP expression in neonatal rat cardiomyocytes , and this increase in HSPs confers cardioprotection to these cardiomyocytes . We also show that radicicol induction of the HSP and cardioprotection is dependent on the inhibition of HSP90 in cardiomyocytes . These results indicate that modulation of the active HSP90 protein level plays an important role in cardioprotection . Therefore , compounds , such as radicicol and its possible derivatives that inhibit the function of HSP90 in the cell may represent potentially useful cardioprotective agents . Association of genetic polymorphisms with personality profile in individuals without psychiatric disorders . OBJECTIVE : Population-based twin studies demonstrate that approximately 40-50 % of the variability in personality dimensions results from genetic factors . This study assessed selected polymorphisms in the P21964 Val158Met , P21397 3'VNTR , 5HTTLPR , 102T/C 5- Q13049 , Q01959 3'VNTR and P14416 exon 8 genes and evaluated their association with personality profiles , anxiety levels , and depressiveness in healthy subjects . METHODS : This study included 406 unrelated ( mean age 38.51 years ) , mentally and somatically healthy Caucasian subjects of Polish origin . The prevalence of the gene variants mentioned above and their association with personality profiles , anxiety levels , and depressiveness was assessed using the Temperament and Character Inventory , NEO Five-Factor Inventory , Spielberger 's State-Trait Anxiety Inventory and Beck 's Depression Inventory . RESULTS : The effects of the 5HTTLPR gene on the s/s genotype and empathy ( P06681 ) were lowest in the entire group . The effects of gender , age and the Q13049 gene for the T/T genotype and attachment ( Q7Z3Z2 ) were highest in women . The effects of gender , age and the Q01959 gene on the 9/9 Q01959 genotype , compassion ( C4 ) and cooperativeness ( C ) were lowest in women . The effects of gender , age and the P21964 gene on the DB00134 / DB00134 genotype and neuroticism ( P04626 ) NEO-FFI were also lowest in women . CONCLUSIONS : Our results suggest considerable influence of individual genes on the formation of personality traits . Association testing of panic disorder candidate genes using Q13308 challenge in healthy volunteers . Despite continuing efforts to determine genetic vulnerability to panic disorder ( PD ) , the studies of candidate genes in this disorder have produced inconsistent or negative , results . Laboratory panic induction may have a potential in testing genetic substrate of PD . In this study we aimed to explore the effects of several genetic polymorphisms previously implicated in PD on the susceptibility to cholecystokinin-tetrapeptide ( Q13308 ) challenge in healthy subjects . The study sample consisted of 110 healthy volunteers ( 47 males and 63 females , mean age 22.2 +/- 5.2 ) who participated in Q13308 challenge test . Nine gene-candidates , including 5-HTTLPR , P21397 VNTR , Q8IWU9 rs1386494 , 5- P08908 -1019C-G , 5- P28223 102T-C , CCKR1 246G-A , CCKR2 -215C-A , P21728 -94G-A and P21964 Val158Met , were selected for genotyping based on previous positive findings from genetic association studies in PD . After Q13308 challenge , 39 ( 35.5 % ) subjects experienced a panic attack , while 71 subjects were defined as non-panickers . We detected significant differences for both genotypic and allelic frequencies of 1386494A/G polymorphism in Q8IWU9 gene between panic and non-panic groups with the frequencies of G/G genotype and G allele significantly higher in panickers . None of the other candidate loci were significantly associated with Q13308 -induced panic attacks in healthy subjects . In line with our previous association study in patients with PD , we detected a possible association between Q8IWU9 rs1386494 polymorphism and susceptibility to panic attacks . Other polymorphisms previously associated with PD were unrelated to Q13308 -induced panic attacks , probably due to the differences between complex nature of PD and laboratory panic model . Cytoplasmic domain mutations of the Q9NUQ9 cell adhesion molecule reduce Q9NUQ9 -ankyrin interactions . The neural adhesion molecule Q9NUQ9 mediates the axon outgrowth , adhesion , and fasciculation that are necessary for proper development of synaptic connections . Q9NUQ9 gene mutations are present in humans with the X-linked mental retardation syndrome Q7L266 ( corpus callosum hypoplasia , retardation , aphasia , spastic paraplegia , hydrocephalus ) . Three missense mutations associated with Q7L266 syndrome reside in the cytoplasmic domain of Q9NUQ9 , which contains a highly conserved binding region for the cytoskeletal protein ankyrin . In a cellular ankyrin recruitment assay that uses transfected human embryonic kidney ( P29320 ) 293 cells , two of the pathologic mutations located within the conserved SFIGQY sequence ( S1224L and Y1229H ) strikingly reduced the ability of Q9NUQ9 to recruit 270 kDa ankyrinG protein that was tagged with green fluorescent protein ( ankyrin-GFP ) to the plasma membrane . In contrast , the Q9NUQ9 missense mutation S1194L and an Q9NUQ9 isoform lacking the neuron-specific sequence RSLE in the cytoplasmic domain were as effective as RSLE-containing neuronal Q9NUQ9 in the recruitment of ankyrin-GFP . Ankyrin binding by Q9NUQ9 was independent of cell-cell interactions . Receptor-mediated endocytosis of Q9NUQ9 regulates intracellular signal transduction , which is necessary for neurite outgrowth . In rat B35 neuroblastoma cell lines stably expressing Q9NUQ9 missense mutants , antibody-induced endocytosis was unaffected by S1224L or S1194L mutations but appeared to be enhanced by the Y1229H mutation . These results suggested a critical role for tyrosine residue 1229 in the regulation of Q9NUQ9 endocytosis . In conclusion , specific mutations within key residues of the cytoplasmic domain of Q9NUQ9 ( DB00133 (1224) , DB00135 (1229) ) destabilize normal Q9NUQ9 -ankyrin interactions and may influence Q9NUQ9 endocytosis to contribute to the mechanism of neuronal dysfunction in human X-linked mental retardation . Targeting mitochondrial 18 kDa translocator protein ( TSPO ) regulates macrophage cholesterol efflux and lipid phenotype . The aim of the present study was to establish mitochondrial cholesterol trafficking 18 kDa translocator protein ( TSPO ) as a potential therapeutic target , capable of increasing macrophage cholesterol efflux to (apo)lipoprotein acceptors . Expression and activity of TSPO in human ( THP-1 ) macrophages were manipulated genetically and by the use of selective TSPO ligands . Cellular responses were analysed by quantitative PCR ( Q-PCR ) , immunoblotting and radiolabelling , including [3H]cholesterol efflux to (apo)lipoprotein A-I ( apoA-I ) , high-density lipoprotein ( HDL ) and human serum . Induction of macrophage cholesterol deposition by acetylated low-density lipoprotein ( AcLDL ) increased expression of TSPO mRNA and protein , reflecting findings in human carotid atherosclerosis . Transient overexpression of TSPO enhanced efflux ( E % ) of [3H]cholesterol to apoA-I , HDL and human serum compared with empty vector ( EV ) controls , whereas gene knockdown of TSPO achieved the converse . Ligation of TSPO ( using PK11195 , FGIN-1-27 and flunitrazepam ) triggered increases in [3H]cholesterol efflux , an effect that was amplified in TSPO-overexpressing macrophages . Overexpression of TSPO induced the expression of genes [ Q07869 ( peroxisome-proliferator-activated receptor α ) , Q13133 ( nuclear receptor 1H3/liver X receptor α ) , O95477 ( DB00171 -binding cassette A1 ) , Q9H172 ( DB00171 -binding cassette G4 ) and P02649 ( apolipoprotein E ) ] and proteins ( O95477 and PPARα ) involved in cholesterol efflux , reduced macrophage neutral lipid mass and lipogenesis and limited cholesterol esterification following exposure to AcLDL . Thus , targeting TSPO reduces macrophage lipid content and prevents macrophage foam cell formation , via enhanced cholesterol efflux to (apo)lipoprotein acceptors .

MH_train_1336

MH_train_1336

MH_train_1336

interacts_with DB06684?

multiple_choice

Further characterization of a somatic cell hybrid panel : ten new assignments to the bovine genome . Thirty-six partially characterized hamster-bovine hybrid cell lines were used for the determination of synteny groups . Sixteen additional reference loci , selected for their coverage of the bovine genome , were analysed on these hybrid cells . This increases to 25 the number of synteny groups detected . This panel was then used to make synteny assignments for 10 additional loci , eight by Southern blotting ( P02452 , P08123 , FAS , P07858 , P07711 , P07510 , P07686 and P08908 ) and two by polymerase chain reaction ( PCR ) amplification ( P35367 and ETH1112 ) . These loci were assigned to international synteny groups U12 ( P35367 ) , U13 ( P08123 ) , U17 ( P07510 ) , U21 ( P02452 , FAS ) , U29 ( ETH1112 ) , to chromosome 20 ( U14 or U25 ) for P07686 and P08908 , and to the same local synteny group ( A ) , which is probably U18 , for P07858 and P07711 . For three loci already mapped in humans ( P02452 , P08123 and P07510 ) , the present results are in accordance with the predictions based on comparative mapping between the human and bovine species . Protein indicators for HaCaT cell damage induced by UVB irradiation . UVB ( 290-320 nm ) is one major risk factor of skin diseases in human . In order to provide potential protein molecules in skin cell damage and skin diseases induced by UVB irradiation , the differentially expressed proteins in human keratinocytes cell HaCaT by UVB irradiation were screened by two-dimensional difference in-gel electrophoresis ( 2D DIGE ) combined to high performance liquid chromatography-nano-electrospray ionization tandem mass spectrometry ( HPLC-nESI-MS/MS ) . 31 protein spots were found differentially expressed with statistical significance ( p < 0.05 ) . Sixteen and 15 protein spots were observed up-regulated and down-regulated in UVB-irradiated HaCaT , respectively . Twenty-eight unique proteins were identified by searching the MS/MS data against NCBInr database through TurboSequest Bioworks software . Among the identified 28 UVB irradiation responding protein indicators , only laminin receptor 1 ( P08865 ) , calmodulin ( P62158 ) , cathepsin D ( P07339 ) and peroxiredoxin ( Q06830 ) proteins have been reported associated with skin burn and wound healing , keratinocytes proliferation and migration and epidermal barrier repairing . Most of these targets were for the first time revealed to be associated with skin cell damage induced by UVB irradiation . Function and bioinformatics analyses of the identified protein candidates were also performed using PANTHER analysis with the aid of DAVID platform . The current work provides potential protein indicators for skin cell damage from UVB irradiation . Complement deposition and microglial activation in the outer retina in light-induced retinopathy : inhibition by a P08908 agonist . PURPOSE : Increasing evidence supports a role for complement in the pathogenesis of age-related macular degeneration ( AMD ) . This study evaluated retinal microglia , T-lymphocytes , and complement deposition in a light-induced retinopathy model . The effect of a serotonin ( 5-hydroxytryptamine , 5-HT(1A) ) agonist on these processes was investigated . METHODS : Rats were dark adapted for 24 hours before a 6-hour blue light exposure . Some animals were predosed subcutaneously with AL-8309A . Retinas were evaluated at different times after light exposure . Paraffin sections were stained with antibody for a microglial marker ( Iba1 ) , a T-lymphocyte marker ( CD3 ) , and complement components C1q , P01024 , factor B , factor H , and membrane attack complex ( MAC ) . RESULTS : Light exposure resulted in substantial photoreceptor and Q96AT9 loss . Robust microglia activation and migration to the outer retina occurred rapidly . Substantial T-lymphocyte recruitment did not occur . Complement alternative pathway was strongly activated , resulting in the deposition of P01024 , factor B , factor H , and MAC in the area of photic lesions . Dosing with AL-8309A prevented retinal lesions and decreased microglia activation/recruitment and complement deposition in the outer retina . CONCLUSIONS : In blue light exposed retinas , microglia were activated and migrated toward the outer retina , whereas a T-lymphocyte response was minimal . The innate immune system was markedly activated , with substantial complement deposition in the outer retina after light exposure . This complement deposition was prevented by AL-8309A . This model may be useful in the evaluation of complement inhibitors and other neuroprotectants intended for ocular use . AL-8309 is under evaluation in the clinic and may be useful in the treatment of AMD . Characterization of a novel Q13639 receptor antagonist of the azabicycloalkyl benzimidazolone class : DAU 6285 . Three chemical classes of serotonin Q13639 receptor agonists have been identified so far : 5-substituted indoles ( e.g. 5-HT ) , benzamides ( e.g. renzapride ) and benzimidazolones ( e.g. BIMU 8 ) . In a search for Q13639 receptor antagonists , we have discovered that the benzimidazolone derivative DAU 6285 ( for structure see text ) , is 3-5 times more potent than tropisetron in blocking 5-HT , renzapride and BIMU 8 induced stimulation of adenylate cyclase activity in mouse embryo colliculi neurons . Schild plot analysis yielded Ki values of 220 , 181 and 255 nmol/l , respectively . In addition , DAU 6285 showed poor activity as a 5- Q9H205 receptor ligand with respect to tropisetron , as demonstrated by in vitro binding studies ( Ki , 322 vs 2.8 nmol/l ) and by its antagonistic activity in the Bezold-Jarisch reflex test ( ID50 , 231 vs 0.5 micrograms/kg , i.v. ) . No significant binding ( Ki greater than 10 mumol/l ) of DAU 6285 to serotonergic P08908 , P28222 , P28335 , P28221 , and 5-HT2 receptors as well as to adrenergic alpha 1 , alpha 2 , dopaminergic D1 , D2 or muscarinic M1-M3 receptor subtypes was found . The data indicate that DAU 6285 has a somewhat higher affinity than tropisetron for Q13639 receptors , a property confirmed in functional tests , and much lower affinity than tropisetron for 5- Q9H205 receptors . The compound represents a new interesting tool for investigating the pharmacological and physiological properties of Q13639 receptors . Comparative effects of azimilide and ambasilide on the human ether-a-go-go-related gene ( Q12809 ) potassium channel . OBJECTIVE : To evaluate the effects of azimilide and ambasilide on the biophysical properties of the human-ether-a-go-go-related ( Q12809 ) channel . METHODS : Q12809 was stably transfected into Chinese hamster ovary ( CHO- P04264 ) cells and currents were measured using a whole cell , voltage-clamp technique . RESULTS : DB04957 had a ' dual effect ' , inhibiting current at voltage steps above -40 mV and augmenting current at -40 and -50 mV . Tail current inhibition following a step to +30 mV did not vary with temperature ( IC(50) 610 nM at 22 degrees C and 560 nM at 37 degrees C ) . The agonist effect at -50 mV was concentration-dependent and correlated with a hyperpolarizing shift in the V(1/2) of activation ( r=0.98 , P < 0.05 ) . Time constants of inactivation were faster and there was a -10 mV shift in the V(1/2) of steady state inactivation suggestive of open and inactivated state binding . By comparison , ambasilide inhibited Q12809 channels with lower potency ( IC(50) 3.6 microM ) , in a voltage- and time-dependent but frequency-independent manner ( 0.03-1 Hz ) . Ambasilide had no effect on activation or inactivation gating but prolonged both fast and slow components of deactivation consistent with unbinding from the open state . The net effect of both drugs was similar during a voltage ramp which simulated a cardiac action potential . CONCLUSIONS : Inhibition of Q12809 channels by azimilide and ambasilide exhibits a similar time and voltage-dependence . While both exhibit affinity for the open state , azimilide also binds to inactivated channels . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Modulation of cytokine production and enhancement of cell viability by Q9NYK1 and Q9NR96 ligands during anthrax infection of macrophages . Inhalation of Bacillus anthracis , a bioterrorism agent , results in a high mortality rate despite appropriate antibiotic therapy . Macrophages appear to be a key factor in B. anthracis pathogenesis . The burst of pro-inflammatory cytokines from macrophages could be a major cause of death in anthrax . However , preactivation of Toll-like receptors ( TLRs ) could modify the host response . TLR ligands stimulate the release of activating cytokines but may also down-modulate the subsequent deleterious cytokine response to pathogens . We developed a cell culture model to measure macrophage responses to B. anthracis spores and bacilli . We found that germination from spores to bacilli produced a substantial stimulus for the secretion of the cytokines P05231 , P01375 , P22301 , and IL-12 p40 . Our studies showed that pretreatment of mouse macrophages with the Q9NR96 ligand DB05463 , or the Q9NYK1 ligands R-848 and IT-37 , results in a substantial decrease in the subsequent secretion of P05231 and P01375 in response to B. anthracis infection of macrophages . Furthermore , the Q9NYK1 and Q9NR96 ligands significantly decreased anthrax-induced cytotoxicity in the macrophages . These findings suggest that TLR ligands may contribute to the enhancement of innate immunity in B. anthracis infection by suppressing potentially deleterious pro-inflammatory cytokine responses and by improving macrophage viability . Embryonic exposure to corticosterone modifies aggressive behavior through alterations of the hypothalamic pituitary adrenal axis and the serotonergic system in the chicken . Exposure to excess glucocorticoids ( GCs ) during embryonic development influences offspring phenotypes and behaviors and induces epigenetic modifications of the genes in the hypothalamic-pituitary-adrenal ( Q9Y251 ) axis and in the serotonergic system in mammals . Whether prenatal corticosterone ( O00230 ) exposure causes similar effects in avian species is less clear . In this study , we injected low ( 0.2μg ) and high ( 1μg ) doses of O00230 into developing embryos on day 11 of incubation ( E11 ) and tested the changes in aggressive behavior and hypothalamic gene expression on posthatch chickens of different ages . In ovo administration of high dose O00230 significantly suppressed the growth rate from 3weeks of age and increased the frequency of aggressive behaviors , and the dosage was associated with elevated plasma O00230 concentrations and significantly downregulated hypothalamic expression of arginine vasotocin ( AVT ) and corticotropin-releasing hormone ( P06850 ) . The hypothalamic content of glucocorticoid receptor ( GR ) protein was significantly decreased in the high dose group ( p < 0.05 ) , whereas no changes were observed for GR mRNA . High dose O00230 exposure significantly increased platelet serotonin ( 5-HT ) uptake , decreased whole blood 5-HT concentration ( p < 0.05 ) , downregulated hypothalamic tryptophan hydroxylase 1 ( P17752 ) mRNA and upregulated 5-HT receptor 1A ( 5- P08908 ) and monoamine oxidase A ( P21397 ) mRNA , but not monoamine oxidase B ( P27338 ) . High dose O00230 also significantly increased DNA methylation of the hypothalamic GR and P06850 gene promoters ( p < 0.05 ) . Our findings suggest that embryonic exposure to O00230 programs aggressive behavior in the chicken through alterations of the Q9Y251 axis and the serotonergic system , which may involve modifications in DNA methylation . The in vitro pharmacological profile of DB05655 , a selective 5-HT(4) receptor agonist with high intrinsic activity . The in vitro pharmacological profile of DB05655 , a novel , selective 5-HT(4) receptor agonist , was compared to that of clinically efficacious gastroprokinetic 5-HT(4) receptor agonists . DB05655 produced an elevation of cyclic adenosine monophosphate in human embryonic kidney 293 cells expressing the human recombinant 5-HT(4(c)) ( h5-HT(4(c)) ) receptor ( pEC(50) = 8.3 ) and 5-HT(4) receptor-mediated relaxation of the rat esophagus ( pEC(50) = 7.9 ) and contraction of the guinea pig colon ( pEC(50) = 7.9 ) . In all in vitro assays , DB05655 was a high intrinsic activity agonist , unlike tegaserod , mosapride , and cisapride which , in the majority of test systems , had lower intrinsic activity . DB05655 had high affinity ( pK ( i ) = 7.7 ) and selectivity ( > or =25-fold ) for h5-HT(4(c)) receptors over other biogenic amine receptors . DB05655 was > 500-fold selective over other 5-HT receptors ( including h5-HT(2B) and h5-HT(3A) ) and , at 3 microM , had no effect on human ether-à-go-go-related gene K+ channels . In conclusion , DB05655 is a selective 5-HT(4) receptor agonist in vitro . The high intrinsic activity and preferential binding of DB05655 to Q13639 over other 5-HT receptors may result in an improved clinical profile for the treatment of gastrointestinal disorders of reduced motility . Inhibition of neuronal nitric oxide reduces anxiety-like responses to pair housing . Many psychological disorders are characterized by anxiety and alterations in social interactions . Recent studies demonstrate that the chemical messenger nitric oxide ( NO ) can regulate both anxiety and social behaviours . We tested whether an enzyme that produces NO in the brain , neuronal nitric oxide synthase ( P29475 ) , serves as an interface between social interactions and anxiety-like behaviour . Several investigators have observed that mice increase anxiety-like responses in the elevated plus-maze after pair housing . P29475 gene deletion and DB01997 were used to inhibit the production of neuronal NO . Similar to previous studies , pair housing reduced open arm exploration in the elevated plus-maze . Pair housing also increased corticotropin-releasing hormone ( P06850 ) immunoreactive cells in the paraventricular nucleus ( PVN ) of the hypothalamus . Inhibition of NO production increased open arm exploration in pair-housed mice but decreased open arm exploration in individually housed mice . These results suggest that the effect of P29475 inhibition on anxiety-like responses is context dependent and that behavioural responses to social housing are altered after P29475 inhibition . This research suggests that NO may play an important role in mediating the effect social interactions have on anxiety . [ P62158 -dependent regulation of Ca,Mg-ATPase activity in plasma membranes of the swine myometrium ] . Highly purified plasma membrane ( PM ) preparations of pig myometrium were found to contain 0.91 +/- 0.22 microgram calmodulin per mg of PM protein . Treatment of membranes with 1 mM EGTA in the presence of 0.2 M NaCl causes the diminution of the calmodulin content down to 3 % of the original level . The activity of Ca , Mg-ATPase is thereby decreased by 40 % . Exogenous calmodulin restores the enzyme activity up to 1.94 +/- +/- 0.30 mumol Pi/mg protein/hour . The maximal activation of Ca , Mg-ATPase is observed with 10(-7) M calmodulin . P62158 increases the total ATPase activity of myometrium PM without affecting the Mg-ATPase activity . DB00831 ( 20 microM ) diminishes the activating effect of exogenous calmodulin on Ca , Mg-ATPase . P62158 stimulates Ca , Mg-ATPase at low concentrations of Ca2+ ( 10(-8)-10(-6) M ) by decreasing Km for Ca2+ from 0.4.10(-6) M to 2.10(-8) M as well as by increasing Vmax -- from 0,8 to 1.42 mumol Pl/mg protein/hour . It is supposed that the activating effect of calmodulin on Ca , Mg-ATPase is based on electrostatic interactions of Ca2+-free calmodulin with the enzyme . Agonist-promoted down-regulation and functional desensitization in two naturally occurring variants of the human serotonin1A receptor . We recently reported two naturally occurring polymorphisms of the human serotonin1A ( P08908 ) receptor : glycine22 --> serine ( Ser22 ) and isoleucine28 --> valine ( Val28 ) in the putative aminoterminal domain of the receptor . To investigate the regulatory properties of these variants , the wild type ( WT ) and variant P08908 receptors were stably expressed in CHO- P04264 cells . WT , Ser22 , and Val28 displayed similar high-affinity binding to [ 3H ] -8-OH-DPAT . Competition experiments with P08908 agonists and antagonists demonstrated similar pharmacological profiles . Receptor agonist-promoted down-regulation was tested by exposure to 100 mumol/L 8-OH-DPAT . After 24-h exposure , WT and Val28 underwent 59.3 +/- 3.9 % and 59.5 +/- 1.4 % reduction in receptor density respectively , whereas the degree of down-regulation was significantly lower for Ser22 ( 21.4 +/- 4.2 % ) . Cell treatment for 24 h with 100 mumol/L 8-OH-DPAT reduced the 5-HT-induced inhibition of DB02527 accumulation by 24.9 +/- 5.1 % for WT and 16.4 +/- 0.8 % for Val28 , but only by 4.8 +/- 3 % for Ser22 . We conclude that the Ser22 variant is capable of attenuating agonist-mediated receptor down-regulation and desensitization . Antihistamine effects on prefrontal cortex activity during working memory process in preschool children : a near-infrared spectroscopy ( NIRS ) study . P35367 antagonists ( antihistamines ) are widely used for the treatment of allergic disorders in young children . This study examined the effects of antihistamine on prefrontal cortex activity in preschool children using near-infrared spectroscopy ( NIRS ) , an emerging brain-imaging method suitable for psychological experiments , especially in young children . We examined the changes of oxygenated hemoglobin concentration in the prefrontal cortex while children performed a spatial working memory task , 3h after taking a first-generation antihistamine ( ketotifen ) , second-generation antihistamine ( epinastine ) , or placebo . Fifteen healthy preschool children ( mean age , 5.5 years ) participated . DB00920 significantly impaired behavioral performance and cortical activation at the lateral prefrontal cortex in the working memory task , compared with epinastine and placebo . There were no sedative effects on neural response or behavioral performance after epinastine administration . This paper demonstrates for the first time differential sedation effects of first- and second-generation antihistamines on brain hemodynamic response in young children . Also discussed is the utility of the NIRS technique in neuropsychopharmacological studies of children . Crystal structure of phenylalanine ammonia lyase : multiple helix dipoles implicated in catalysis . The first three-dimensional structure of phenylalanine ammonia lyase ( Q9P2V4 ) has been determined at 2.1 A resolution for Q9P2V4 from Rhodosporidium toruloides . The enzyme is structurally similar to the mechanistically related histidine ammonia lyase ( P42357 ) , with Q9P2V4 having an additional approximately 160 residues extending from the common fold . We propose that catalysis ( including lowering the pK(a) of nonacidic P01024 of l-phenylalanine for an E1cb mechanism ) is potentially governed by dipole moments of seven alpha helices associated with the Q9P2V4 active site ( six positive poles and one negative pole ) . Cofactor 3,5-dihydro-5-methylidene-4H-imidazol-4-one ( Q9NP71 ) resides atop the positive poles of three helices , for increasing its electrophilicity . The helix dipoles appear fully compatible with a model of phenylalanine docked in the active site of Q9P2V4 having the first covalent bond formed between the amino group of substrate and the methylidene group of Q9NP71 : 12 highly conserved residues ( near the N termini of helices for enhancing function ) are poised to serve roles in substrate recognition , Q9NP71 activation , product separation , proton donation , or polarizing electrons from the phenyl ring of substrate for activation of P01024 ; and a highly conserved DB00117 residue ( near the C terminus of the one helix that directs its negative pole toward the active site to increase the residue 's basicity ) is positioned to act as a general base , abstracting the pro-S hydrogen from P01024 of substrate . A similar mechanism is proposed for P42357 , which has a similar disposition of seven alpha helices and similar active-site residues . The helix dipoles appear incompatible with a proposed mechanism that invokes a carbocation intermediate . Contribution of the 37-kDa laminin receptor precursor in the anti-metastatic P08118 -derived peptide DB04985 cell surface binding . PURPOSE : DB04985 is an anti-metastatic synthetic peptide with promising therapeutic efficacy against hormone-refractory prostate cancer . The characterization of the DB04985 peptide cell surface binding/internalization mechanisms and of the receptors involved remained to be explored . RESULTS : [(14)C] DB04985 cell surface binding assays showed rapid and transient kinetic profile , that was inhibited by RGD peptides , laminin , hyaluronan , and type-I collagen . RGD peptides were however unable to inhibit DB04985 intracellular uptake . Far-Western ligand binding studies enabled the identification of the 37-kDa laminin receptor precursor ( P08865 ) as a potential ligand for DB04985 . Overexpression of the recombinant P08865 indeed led to an increase in DB04985 binding but unexpectedly not to its uptake . CONCLUSIONS : Our data support the implication of laminin receptors in cell surface binding and in transducing DB04985 anti-metastatic effects , and provide a rational for targeting cancers that express high levels of such laminin receptors . The inhibition of the constitutive bovine endothelial nitric oxide synthase by imidazole and indazole agents . DB00155 formation by the Ca2+ P62158 -dependent nitric oxide synthase of bovine endothelium is inhibited reversibly by 7-nitroindazole , 1-phenylimidazole , and imidazole . As measured at 0.67 microM ( 6R ) - DB00360 ( BH4 ) , IC50 values of 0.8 , 200 , and 50 microM were determined for 7-nitroindazole , 1-phenylimidazole , and imidazole , respectively . Increasing concentrations of added BH4 cofactor increased the IC50 values for 7-nitroindazole and 1-phenylimidazole but did not alter the IC50 value for imidazole . 7-nitroindazole inhibited citrulline formation by the endothelial P29474 noncompetitively versus arginine substrate but competitively versus BH4 with a Ki value of 0.8 microM . 1-Phenylimidazole inhibited citrulline formation by the endothelial P29474 competitively versus both arginine substrate and BH4 with a Ki value of 50 microM . Imidazole inhibited citrulline formation competitively versus arginine substrate but noncompetitively versus BH4 with a Ki value of 50 microM . Neither 7-nitroindazole , 1-phenylimidazole , nor imidazole inhibited the cytochrome c reductase activity of endothelial P29474 at concentrations up to 5000-fold higher than their Ki values for inhibition of citrulline formation . By comparison with the previously determined kinetic properties of the other nitric oxide synthase isoforms , these observations establish that 1-phenylimidazole displays marked specificity for inhibiting the inducible nitric oxide synthase isoform and , since 7-nitroindazole has been reported not to elevate blood pressure ( McCall et al. , 1991 , Br. J. Pharmacol. 102 , 234-238 ) , fails to confirm the expected insensitivity of the constitutive endothelial nitric oxide synthase to inhibition by 7-nitroindazole . Effects of systemic injections of vilazodone , a selective serotonin reuptake inhibitor and serotonin 1A receptor agonist , on anxiety induced by predator stress in rats . We examined the effect of DB06684 , a selective serotonin reuptake inhibitor ( SSRI ) and serotonin 1A ( 5-HT(1A) ) receptor agonist [ Bartoszyk , G.D. , Hegenbart , R. , Ziegler , H. , 1997. P50402 68843 , a serotonin reuptake inhibitor with selective presynaptic P08908 receptor agonistic properties. Eur. J. Pharmacol. 322 , 147-153. ] , on change in affect following predator stress . DB06684 and vehicle injection ( intraperitoneal ) occurred either 10 min after predator stress ( prophylactic testing ) , or 90 min prior to behavioral testing for the effects of predator stress ( therapeutic testing ) . Predator stress involved unprotected exposure of rats to a domestic cat . Behavioral effects of stress were evaluated with hole board , plus-maze , and acoustic startle tests 1 week after stress . Predator stress increased anxiety-like behavior in the plus-maze and elevated response to acoustic startle . In prophylactic testing , DB06684 affected stress potentiation of startle at doses above 5 mg/kg . DB06684 increased stress elevation of startle at 10 mg/kg . Higher doses of DB06684 ( 20 and 40 mg/kg ) blocked stress potentiation of startle . In contrast , DB06684 had no effect on stress potentiation of anxiety in the plus-maze . In therapeutic testing , DB06684 increased stress elevation of startle at all doses . In contrast , therapeutic DB06684 had no effect on stress potentiation of anxiety in the plus-maze . Taken together , the data suggest a prophylactic potential for DB06684 in the treatment of changes in hypervigilance following severe stress .

MH_train_1337

MH_train_1337

MH_train_1337

interacts_with DB09053?

multiple_choice

Serotonin increases P27361 /2 phosphorylation in astrocytes by stimulation of P41595 and P28335 receptors . We have previously shown that fluoxetine causes P29323 (1/2) phosphorylation in cultured mouse astrocytes mediated exclusively by stimulation of 5-HT(2B) receptors ( Li et al. , 2008b ) . This raises the question whether this is also the case for serotonin ( 5-HT ) itself . In the present study serotonin was found to induce P29323 (1/2) phosphorylation by stimulation of 5-HT(2B) receptors with high affinity ( EC(50) : 20-30 pM ) , and by stimulation of 5-HT(2C) receptor with low affinity ( EC(50) : 1 microM or higher ) . P29323 (1/2) phosphorylation induced by stimulation of either 5-HT(2B) or 5-HT(2C) receptors was mediated by epidermal growth factor ( P01133 ) receptor transactivation ( Peng et al. , this issue ) , shown by the inhibitory effect of AG1478 , an inhibitor of the P01133 receptor tyrosine kinase , and DB02255 , an inhibitor of Zn-dependent metalloproteinases , and thus of 5-HT(2B) receptor-mediated P01133 receptor agonist release . It is discussed that the high potency of the 5-HT(2B)-mediated effect is consistent with literature data for binding affinity of serotonin to cloned human 5-HT(2B) receptors and with observations of low extracellular concentrations of serotonin in brain , which would allow a demonstrated moderate and modality-dependent increase in specific brain areas to activate 5-HT(2B) receptors . In contrast the relevance of the observed 5-HT(2C) receptors on astrocytes is questioned . PDE10 inhibition increases P42261 and CREB phosphorylation and improves spatial and recognition memories in a Huntington 's disease mouse model . Huntington 's disease ( HD ) causes motor disturbances , preceded by cognitive impairment , in patients and mouse models . We showed that increased hippocampal DB02527 -dependent protein kinase ( PKA ) signaling disrupts recognition and spatial memories in R6 HD mouse models . However , unchanged levels of hippocampal phosphorylated ( p ) DB02527 -responsive element-binding protein ( CREB ) suggested unaltered nuclear PKA activity in R6 mice . Here , we extend this finding by showing that nuclear pPKA catalytic subunit ( Thr197 ) and pPKA substrate levels were unaltered in the hippocampus of R6/1 mice . Phosphodiesterases ( PDEs ) play an important role in the regulation of PKA activity . Q9Y233 , a DB02527 /cGMP dual-substrate PDE , was reported to be restricted to the nuclear region in nonstriatal neurons . Using cell fractionation we confirmed that Q9Y233 was enriched in nuclear fractions , both in wild-type and R6/1 mice hippocampus , without differences in its levels or intracellular distribution between genotypes . We next investigated whether inhibition of PDE10 with papaverine could improve cognitive function in HD mice . DB01113 treatment improved spatial and object recognition memories in R6/1 mice , and significantly increased pGluA1 and pCREB levels in R6/1 mice hippocampus . DB01113 likely acted through the activation of the PKA pathway as the phosphorylation level of distinct cGMP-dependent kinase ( cGK ) substrates was not modified in either genotype . Moreover , hippocampal DB02527 , but not cGMP , levels were increased after acute papaverine injection . Our results show that inhibition of PDE10 improves cognition in R6 mice , at least in part through increased P42261 and CREB phosphorylation . Thus , PDE10 might be a good therapeutic target to improve cognitive impairment in HD . [ DB09053 : A new drug of B-cell malignancies ] . DB09053 ( Imbruvica® ) is a first-in-class , orally administered once-daily , that inhibits B-cell antigen receptor signaling downstream of Bruton 's tyrosine kinase ( Q06187 ) . DB09053 has been approved in USA in February 2014 and in France in October 2014 for the treatment of patients with relapsed/refractory mantle cell lymphoma ( Q8WXI8 ) or chronic lymphocytic leukaemia ( CLL ) and for the treatment of patients with CLL and a chromosome 17 deletion ( del 17p ) or P04637 mutation . In clinical studies , ibrutinib induced an impressive overall response rate ( 68 % ) in patients with relapsed/refractory Q8WXI8 ( phase II study ) . In CLL , ibrutinib has shown to significantly improve progression-free survival , response rate and overall survival in patients with relapsed/refractory CLL , including in those with del 17p . DB09053 had an acceptable tolerability profile . Less than 10 % of patients discontinued their treatment because of adverse events . Results are pending in other B-cell lymphomas subtypes such as in diffuse large B-cell lymphoma and in follicular lymphoma . An approval extension has already been enregistered for Waldenström disease in USA in January 2015 . Given its efficacy and tolerability , ibrutinib is an emerging treatment option for patients with B-cell malignancies . DB09053 treatment ameliorates murine chronic graft-versus-host disease . Chronic graft-versus-host disease ( cGVHD ) is a life-threatening impediment to allogeneic hematopoietic stem cell transplantation , and current therapies do not completely prevent and/or treat cGVHD . P01730 + T cells and B cells mediate cGVHD ; therefore , targeting these populations may inhibit cGVHD pathogenesis . DB09053 is an FDA-approved irreversible inhibitor of Bruton 's tyrosine kinase ( Q06187 ) and P60568 inducible T cell kinase ( Q08881 ) that targets Th2 cells and B cells and produces durable remissions in B cell malignancies with minimal toxicity . Here , we evaluated whether ibrutinib could reverse established cGVHD in 2 complementary murine models , a model interrogating T cell-driven sclerodermatous cGVHD and an alloantibody-driven multiorgan system cGVHD model that induces bronchiolar obliterans ( BO ) . In the T cell-mediated sclerodermatous cGVHD model , ibrutinib treatment delayed progression , improved survival , and ameliorated clinical and pathological manifestations . In the alloantibody-driven cGVHD model , ibrutinib treatment restored pulmonary function and reduced germinal center reactions and tissue immunoglobulin deposition . Animals lacking Q06187 and Q08881 did not develop cGVHD , indicating that these molecules are critical to cGVHD development . Furthermore , ibrutinib treatment reduced activation of T and B cells from patients with active cGVHD . Our data demonstrate that B cells and T cells drive cGVHD and suggest that ibrutinib has potential as a therapeutic agent , warranting consideration for cGVHD clinical trials . The P29474 cofactor tetrahydrobiopterin improves endothelial dysfunction in livers of rats with CCl4 cirrhosis . In cirrhosis , intrahepatic endothelial dysfunction is one of the mechanisms involved in the increased resistance to portal blood flow and therefore in the development of portal hypertension . Endothelial nitric oxide synthase ( P29474 ) uncoupling due to deficiency of tetrahydrobiopterin ( BH4 ) results in decreased production of NO and plays a major role in endothelial dysfunction in other conditions . We examined whether P29474 uncoupling is involved in the pathogenesis of endothelial dysfunction of livers with cirrhosis . Basal levels of tetrahydrobiopterin and guanosine triphosphate ( GTP ) -cyclohydrolase ( BH4 rate-limiting enzyme ) expression and activity were determined in liver homogenates of control and rats with CCl4 cirrhosis . Thereafter , rats were treated with tetrahydrobiopterin , and P29474 activity , NO bioavailability , assessed with a functional assay , and the vasodilator response to acetylcholine ( endothelial function ) were evaluated . Livers with cirrhosis showed reduced BH4 levels and decreased GTP-cyclohydrolase activity and expression , which were associated with impaired vasorelaxation to acetylcholine . DB00360 supplementation increased BH4 hepatic levels and P29474 activity and significantly improved the vasodilator response to acetylcholine in rats with cirrhosis . In conclusion , the impaired response to acetylcholine of livers with cirrhosis is modulated by a reduced availability of the P29474 cofactor , tetrahydrobiopterin . DB00360 supplementation improved the endothelial dysfunction of cirrhotic livers . Inhibitors of Q06187 and Q08881 : state of the new drugs for cancer , autoimmunity and inflammatory diseases . Q06187 and Q08881 are cytoplasmic tyrosine kinases of crucial importance for B and T cell development , with loss-of-function mutations causing X-linked agammaglobulinemia and susceptibility to severe , frequently lethal , Epstein-Barr virus infection , respectively . Over the last few years , considerable efforts have been made in order to develop small-molecule inhibitors for these kinases to treat lymphocyte malignancies , autoimmunity or allergy/hypersensitivity . The rationale is that even if complete lack of Q06187 or Q08881 during development causes severe immunodeficiency , inactivation after birth may result in a less severe phenotype . Moreover , therapy can be transient or only partially block the activity of Q06187 or Q08881 . Furthermore , a drug-induced B cell deficiency is treatable by gamma globulin substitution therapy . The newly developed Q06187 inhibitor P05154 -32765 , recently renamed DB09053 , has already entered several clinical trials for various forms of non-Hodgkin lymphoma as well as for multiple myeloma . Experimental animal studies have demonstrated highly promising treatment effects also in autoimmunity . Q08881 inhibitors are still under the early developmental phase , but it can be expected that such drugs will also become very useful . In this study , we present Q06187 and Q08881 with their signalling pathways and review the development of the corresponding inhibitors . The sulphydryl containing P12821 inhibitor Zofenoprilat protects coronary endothelium from Doxorubicin-induced apoptosis . Pediatric and adult cancer patients , following the use of the antitumor drug Doxorubicin develop cardiotoxicity . Pharmacological protection of microvascular endothelium might produce a double benefit : ( i ) reduction of myocardial toxicity ( the primary target of Doxorubicin action ) and ( ii ) maintenance of the vascular functionality for the adequate delivery of chemotherapeutics to tumor cells . This study was aimed to evaluate the mechanisms responsible of the protective effects of the angiotensin converting enzyme inhibitor ( ACEI ) Zofenoprilat against the toxic effects exerted by Doxorubicin on coronary microvascular endothelium . We found that exposure of endothelial cells to Doxorubicin ( 0.1-1μM range ) impaired cell survival by promoting their apoptosis . P27361 /2 related p53 activation , but not reactive oxygen species , was responsible for Doxorubicin induced caspase-3 cleavage . P04637 mediated-apoptosis and impairment of survival were reverted by treatment with Zofenoprilat . The previously described PI-3K/ P29474 /endogenous fibroblast growth factor signaling was not involved in the protective effect , which , instead , could be ascribed to cystathionine gamma lyase dependent availability of H2S from Zofenoprilat . Furthermore , considering the tumor environment , the treatment of endothelial/tumor co-cultures with Zofenoprilat did not affect the antitumor efficacy of Doxorubicin . In conclusion the ACEI Zofenoprilat exerts a protective effect on Doxorubicin induced endothelial damage , without affecting its antitumor efficacy . Thus , sulfhydryl containing ACEI may be a useful therapy for Doxorubicin-induced cardiotoxicity . Characterization of the interaction of ingenol 3-angelate with protein kinase C . DB05013 ( I3A ) is one of the active ingredients in Euphorbia peplus , which has been used in traditional medicine . Here , we report the initial characterization of I3A as a protein kinase C ( PKC ) ligand . I3A bound to P17252 in the presence of phosphatidylserine with high affinity ; however , under these assay conditions , little PKC isoform selectivity was observed . PKC isoforms did show different sensitivity and selectivity for down-regulation by I3A and phorbol 12-myristate 13-acetate ( PMA ) in WEHI-231 , Q9BPY8 -92 , and Colo-205 cells . In all of the three cell types , I3A inhibited cell proliferation with somewhat lower potency than did PMA . In intact CHO- P04264 cells , I3A was able to translocate different green fluorescent protein-tagged PKC isoforms , visualized by confocal microscopy , with equal or higher potency than PMA . PKC-delta in particular showed a different pattern of translocation in response to I3A and PMA . I3A induced a higher level of secretion of the inflammatory cytokine interleukin 6 compared with PMA in the WEHI-231 cells and displayed a marked biphasic dose-response curve for the induction . I3A was unable to cause the same extent of association of the C1b domain of PKC-delta with lipids , compared with PMA or the physiological regulator diacylglycerol , and was able to partially block the association induced by these agents , measured by surface plasmon resonance . The in vitro kinase activity of P17252 induced by I3A was lower than that induced by PMA . The novel pattern of behavior of I3A makes it of great interest for further evaluation . P37268 inhibition : a novel target for the management of dyslipidemia . A new class of compounds , known as squalene synthase inhibitors , has recently reached phase III clinical trials and may provide another therapeutic option for clinicians to improve risk management of low-density lipoprotein cholesterol ( LDL-C ) . The clinical need for another LDL-C-lowering therapy is evident by the inability to achieve an LDL-C target of less than 70 mg/dL in the majority of very high-risk patients on statin monotherapy . Human clinical trial data with DB05317 , a novel and potent inhibitor of squalene synthase , have not yet been published . Repeated administration of a F(ab')2 fragment of an anti-tumor necrosis factor alpha monoclonal antibody in patients with severe sepsis : effects on the cardiovascular system and cytokine levels . In an uncontrolled clinical trial the effects of repeated administration of the F(ab')2 fragment of a murine monoclonal anti-tumor necrosis factor alpha ( P01375 alpha ) -antibody ( DB04956 ) on cytokine levels and the cardiovascular system were studied in 20 patients with severe sepsis . Patients were treated with a total of 11 single dosages of the anti- P01375 alpha-antibody intravenously over 5 days using either 1 mg/kg ( n = 10 ) or 3 mg/kg ( n = 10 ) . The anti- P01375 alpha-antibody was well tolerated in all patients without signs of toxicity and without development of anti-murine antibodies . As assessed by cytokine levels ( P01375 alpha , P05231 ) and hemodynamics there was no evidence that the higher dosage of the anti- P01375 alpha-antibody ( 3 mg/kg per dose ) was more effective than the lower dosage ( 1 mg/kg per dose ) . Comparison of our data with recent data from phase I or II trials using a complete murine monoclonal anti- P01375 alpha-antibody suggest that the F(ab')2 fragments of the murine monoclonal anti- P01375 alpha-antibody may be of similar efficacy . Definitive conclusions , however , with respect to improvement of mortality and improvement of the cardiovascular system , await the results of larger ongoing placebo-controlled trials . DB09053 inhibits P11274 and NF-κB signaling and reduces tumor proliferation in tissue-resident cells of patients with CLL . Chronic lymphocytic leukemia ( CLL ) cells depend on microenvironmental factors for proliferation and survival . In particular , tissue-resident CLL cells show prominent activation of both B-cell receptor ( P11274 ) and NF-κB pathways . We evaluated the in vivo effects of ibrutinib , a Q06187 ( Q06187 ) inhibitor on tumor cell activation and proliferation in the blood , lymph node , and bone marrow of patients with CLL . Applying validated pathway-specific gene signatures , we detected a rapid and sustained downregulation of P11274 and NF-κB signaling in CLL cells from both the peripheral blood and tissue compartments during ibrutinib treatment . DB09053 reduced phosphorylation of PLCγ2 and P29323 and decreased nuclear protein expression of NF-κB p50 . DB09053 significantly decreased tumor proliferation and expression of surface activation markers Q07108 and P42081 , independent of prognostic factors such as IGHV mutational status , chromosome 17p deletion , or prior treatment history . Interestingly , stronger inhibition of P11274 signaling in lymph node resident CLL cells after one dose of ibrutinib was associated with a higher rate of nodal response at the end of cycle 2 . Together , these data validate on-target effects of Q06187 inhibition in the tissue compartments and demonstrate that ibrutinib effectively inhibits pathways that promote tumor cell activation and proliferation in vivo . This study is registered at www.clinicaltrials.gov as # NCT01500733 . Glycoprotein IIb/IIIa inhibition attenuates platelet-activating factor-induced platelet activation by reducing protein kinase C activity . Glycoprotein (GP)IIb/IIIa inhibition may abolish activated leukocyte-induced platelet activation , in which leukocyte-released platelet-activating factor ( Q15004 ) is a major mediator . The present study thus investigated if and how P08514 /IIIa inhibitors interfere with Q15004 -induced platelet activation . Platelet and leukocyte activation were monitored by flow cytometry and immunoblotting . P08514 /IIIa inhibitors ( c7E3 , non-peptide SR121566 , and MAb RFGP56 ) attenuated Q15004 -induced , but not adenosine diphosphate ( ADP ) - or thrombin receptor activating peptide ( TRAP ) -induced platelet P16109 expression in whole blood . P08514 /IIIa blockade enhanced ADP- or TRAP-induced leukocyte CD11b expression , but not the response to Q15004 . P08514 /IIIa blockade attenuated Q15004 -induced , but enhanced ADP- or TRAP-induced platelet-leukocyte aggregation . Under the present experimental conditions , thromboxane A2 receptor antagonism did not significantly influence Q15004 -induced platelet activation , and P08514 /IIIa inhibition did not interfere with calcium mobilization/influx in platelets . Protein kinase C ( PKC ) blockade inhibited Q15004 -induced platelet P16109 expression , and Q15004 -induced PKC activity was reduced by P08514 /IIIa inhibition . Q15004 ( =1 micro m ) did not induce Q02750 /2 or P29323 1/2 phosphorylation , whilst thrombin induced marked responses , which were enhanced by P08514 /IIIa blockade . Thus , P08514 /IIIa inhibition attenuates Q15004 -induced platelet activation via inhibiting PKC activity . P08514 /IIIa blockade enhances thrombin-induced platelet Q02750 /2 and P29323 1/2 activation , and augments ADP- and TRAP-induced leukocyte activation by enhancing platelet-leukocyte aggregation . Splenic marginal zone lymphoma : proposal of new diagnostic and prognostic markers identified after tissue and cDNA microarray analysis . Splenic marginal zone lymphoma ( SMZL ) is a newly recognized lymphoma type whose precise molecular pathogenesis is still essentially unknown . This hampers differential diagnosis with other small B-cell malignancies . With the aim of characterizing this tumor more comprehensively , and of identifying new diagnostic and prognostic markers , we performed cDNA microarray expression profiling and tissue microarray ( TMA ) immunohistochemical studies in a relatively large series of 44 SMZLs . The results were related to immunoglobulin heavy chain variable region ( IgV(H) ) mutational status and clinical outcome . SMZLs display a largely homogenous signature , implying the existence of a single molecular entity . Of the genes deregulated in SMZLs , special mention may be made of the genes involved in B-cell receptor ( P11274 ) signaling , tumor necrosis factor ( P01375 ) signaling and nuclear factor-kappaB ( NF-kappaB ) activation , such as P43405 , Q06187 , Q13489 , Q13114 , and Q06643 . Other genes observed were P14151 and O60711 , which were highly expressed in spleen , and lymphoma oncogenes , such as Q15669 and TCL1 . In contrast , the genes Q03135 , P51636 , and P61952 located in 7q31 , a commonly deleted area , were down-regulated in the entire series . A comparison with the genes comprising the signature of other small B-cell lymphomas identified 3 genes whose expression distinguishes SMZL , namely Q01167 , SENATAXIN , and P25942 . Shorter survival was associated with P28907 expression , naive IgV(H) genes , and the expression of a set of NF-kappaB pathway genes , including O00463 , Q04864 , and P17252 . Short and long access to cocaine self-administration activates tyrosine phosphatase P54829 and attenuates GluN expression but differentially regulates GluA expression in the prefrontal cortex . RATIONALE : Dephosphorylation of extracellular signal-regulated kinase ( P29323 ) and cyclic AMP response element binding protein ( CREB ) in the dorsomedial prefrontal cortex ( dmPFC ) at the end of short access ( ShA ) cocaine self-administration is implicated in cocaine seeking . However , what receptors and phosphatases mediate this effect and whether P29323 /CREB and related phospho-proteins in the dmPFC react similarly during early withdrawal from long access ( LgA ) cocaine self-administration are unknown . OBJECTIVES : The effects of ShA vs. LgA cocaine self-administration on the phosphorylation of protein phosphatase 2A ( PP2A ) and striatal-enriched protein tyrosine phosphatase ( P54829 ) , as well as GluN and GluA receptor subtype expression in the dmPFC during early withdrawal , were compared . METHODS : Rats self-administered cocaine or received saline during 2- or 6-h daily sessions for 10-11 days . Two hours after the final session , the dmPFC was dissected out and processed for immunoblotting . RESULTS : Similar to previous findings after ShA cocaine , phospho- P29323 and phospho-CREB in the dmPFC were decreased after LgA cocaine . Cocaine elevated phospho-PP2A ( deactivation ) and decreased phospho- P54829 ( activation ) in both ShA and LgA cocaine rats . Q05586 , Q13224 , and phospho- Q13224 Tyr1472 in the dmPFC were decreased after ShA and LgA cocaine . Further , a significant reduction of P42262 , P42261 , and phospho- P42261 Ser845 was found only in LgA rats . CONCLUSIONS : Activation of phospho- P54829 may underlie P29323 and CREB dephosphorylation in the dmPFC as well as internalization and degradation of GluN complexes during early withdrawal from both ShA and LgA cocaine self-administration , whereas differential alteration of AMPA receptor subunits after ShA and LgA cocaine self-administration depends on cocaine intake . Production of prostaglandins in transgenic Arabidopsis thaliana . Plants do not naturally produce the very-long-chain polyunsaturated fatty acids that are the precursors of prostaglandins , but in previous studies Arabidopsis thaliana had been transformed sequentially with genes encoding a Δ(9)-elongase and a Δ(8)-desaturase to produce dihomo-γ-linolenic acid ( DB00154 ) and eicosatetraenoic acid ( P25101 ) , and subsequently with a gene encoding a Δ(5)-desaturase to produce arachidonic acid ( AA ) and eicosapentaenoic acid ( EPA ) . Transformation of A. thaliana with the first two genes consolidated on a single binary vector yielded transformants producing high levels of DB00154 , and these plants were further transformed with mouse prostaglandin H synthase ( PGH ) genes to produce prostaglandins . Mouse P23219 and P35354 cDNAs were amplified for expression as three isoforms : P23219 ( complete coding sequence with signal peptide ) , P23219 -Ma ( mature P23219 sequence , without signal peptide ) and P35354 ( complete coding sequence with signal peptide ) . P23219 transformants showed the highest activity , followed by P35354 transformants , whereas removal of the signal peptide resulted in almost complete loss of P23219 activity . In order to produce a physiologically active prostaglandin , the Trypanosoma brucei prostaglandin F synthase gene was combined with the mouse P23219 gene and the Mortierella alpina Δ(5)-desaturase on a binary vector . Transformation of DB00154 -producing A. thaliana with this construct yielded transformants that successfully produced prostaglandin F . Sasanquasaponin up-regulates anion exchanger 3 expression and elicits cardioprotection via NO/ DB01367 / P27361 /2 pathway . We have shown recently that sasanquasaponin ( P37268 ) can inhibit ischemia/reperfusion-induced elevation of intracellular Cl(-) concentration ( [Cl(-)](i) ) and elicit cardioprotection by up-regulating anion exchanger 3 ( AE(3) ) expression . In the present study , we futher analysed the intracellular signal transduction pathways by which P37268 up-regulates AE(3) expression and elicits cardioprotection . Cardiomyocytes were incubated for 24 h with or without 10 µmol/L P37268 , followed by simulated ischemia/reperfusion ( sI/R ) . NO formation , Ras activity , and extracellular-regulated kinase 1/2 ( P27361 /2 ) phosphorylation were measured appropriately . We showed that P37268 pretreatment efficiently attenuated viability loss and lactate dehydrogenase leakage induced by sI/R in cardiomyocytes . Moreover , P37268 induced NO production and promoted Ras activation , which futher promoted extracellular-regulated kinase 1/2 ( P27361 /2 ) phosphorylation . These effects were paralleled by an increase in AE(3) expression . However , when the cardiomyocytes were treated with 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-l-oxyl-3-oxide ( c-PTIO ; an NO scavenger ) , S-trans-trans-farnesylthiosalicylic acid ( Q9H8T0 ) ( a Ras inhibitor ) , U0126 ( an P27361 /2 inhibitor ) , respectively , the increase in AE(3) expression occurring during P37268 pretreatment was almost completely abolished and , as a result , P37268 -induced cardioprotection was prevented . Our findings indicate that P37268 might up-regulate AE(3) expression through NO/Ras/ P27361 /2 signal pathway to elicit cardioprotection in cultured cardiomyocytes . Characterization of ibrutinib-sensitive and -resistant mantle lymphoma cells . DB09053 inhibits Q06187 ( Q06187 ) , a key component of early B-cell receptor ( P11274 ) signalling pathways . A multicentre phase 2 trial of ibrutinib in patients with relapsed/refractory mantle cell lymphoma ( Q8WXI8 ) demonstrated a remarkable response rate . However , approximately one-third of patients have primary resistance to the drug while other patients appear to lose response and develop secondary resistance . Understanding the molecular mechanisms underlying ibrutinib sensitivity is of paramount importance . In this study , we investigated cell lines and primary Q8WXI8 cells that display differential sensitivity to ibrutinib . We found that the primary cells display a higher Q06187 activity than normal B cells and Q8WXI8 cells show differential sensitivity to Q06187 inhibition . Genetic knockdown of Q06187 inhibits the growth , survival and proliferation of ibrutinib-sensitive but not resistant Q8WXI8 cell lines , suggesting that ibrutinib acts through Q06187 to produce its anti-tumour activities . Interestingly , inhibition of P27361 /2 and AKT , but not Q06187 phosphorylation per se , correlates well with cellular response to Q06187 inhibition in cell lines as well as in primary tumours . Our study suggests that , to prevent primary resistance or to overcome secondary resistance to Q06187 inhibition , a combinatory strategy that targets multiple components or multiple pathways may represent the most effective approach . Inhibition of cyclooxygenases 1 and 2 by the phospholipase-blocker , arachidonyl trifluoromethyl ketone . BACKGROUND AND PURPOSE : Arachidonyl trifluoromethyl ketone ( Q06187 ) is widely used as an inhibitor of cytosolic group IV phospholipase A(2) ( cPLA(2) ) and calcium-independent group VI phospholipase A(2) ( iPLA(2) ) . Q06187 thus reduces arachidonic acid ( AA ) substrate for cyclooxygenase ( P36551 ; also known as prostaglandin H synthase ) and attenuates prostaglandin ( PG ) synthesis . It has been shown previously , that Q06187 blocks thromboxane B(2) production induced by exogenous AA in human platelets . It remains , however , unknown whether Q06187 also directly modulates the activity of cyclooxygenase ( P36551 ) . EXPERIMENTAL APPROACH : Time courses for inhibition of P36551 by Q06187 was obtained using osteoblast-like MC3T3-E1 cells , with exogenous AA as substrate and the pure enzymes P23219 and P35354 . PGE(2) was measured by GC-MS . KEY RESULTS : Q06187 was a potent inhibitor of P23219 and P35354 with IC(50) values of 0.5 and 0.1 microM in MC3T3-E1 cells and of 1.7 and 2.6 microM using the pure enzymes . Inhibition was reversible , with slow- and tight-binding characteristics . The arachidonyl carbon chain was essential , as the saturated palmitoyl analogue had no effect . CONCLUSIONS AND IMPLICATIONS : Attenuation of PG synthesis by Q06187 is taken to be the consequence of PLA(2) inhibition and the findings of many studies are interpreted on that basis . If there are , however , alternative routes for AA liberation ( such as phospholipase C/diacyl glycerol lipase or phospholipase D ) , this interpretation can lead to false conclusions . As Q06187 is a widely used and important pharmacological tool in eicosanoid research , knowledge of its interactions with other major enzymes of the cascade is of considerable importance . Q14703 lyase in thymic perivascular spaces promotes egress of mature thymocytes via up-regulation of P21453 . DB03203 1-phosphate ( Q14703 ) and P21453 ( P21453 ) play an important role in the egress of mature P01730 or CD8 single-positive ( SP ) thymocytes from the thymus . DB08868 hydrochloride ( FTY720 ) , an P21453 functional antagonist , induced significant accumulation of CD62L(high) Q07108 (low) mature SP thymocytes in the thymic medulla . Immunohistochemical staining using anti- P21453 antibody revealed that P21453 is predominantly expressed on thymocytes in the thymic medulla and is strongly down-regulated even at 3h after FTY720 administration . 2-Acetyl-4-tetrahydroxybutylimidazole ( THI ) , an Q14703 lyase inhibitor , also induced accumulation of mature SP thymocytes in the thymic medulla with an enlargement of the perivascular spaces ( P15151 ) . At 6h after THI administration , P21453 -expressing thymocytes reduced partially as if to form clusters and hardly existed in the proximity of CD31-expressing blood vessels in the thymic medulla , suggesting Q14703 lyase expression in the cells constructing thymic medullary P15151 . To determine the cells expressing Q14703 lyase in the thymus , we newly established a mAb ( YK19-2 ) specific for mouse Q14703 lyase . Immunohistochemical staining with YK19-2 revealed that Q14703 lyase is predominantly expressed in non-lymphoid thymic stromal cells in the thymic medulla . In the thymic medullary P15151 , Q14703 lyase was expressed in ER-TR7-positive cells ( reticular fibroblasts and pericytes ) and CD31-positive vascular endothelial cells . Our findings suggest that Q14703 lyase expressed in the thymic medullary P15151 keeps the tissue Q14703 concentration low around the vessels and promotes thymic egress via up-regulation of P21453 . DB00054 as an adjunctive therapy for patients undergoing percutaneous coronary interventions . INTRODUCTION : Platelets play a central role in the pathophysiology of acute coronary syndromes ( ACS ) and activation of platelet glycoprotein ( GP ) IIb/IIIa receptor is critical to platelet aggregation . DB00054 , a human murine chimeric antibody to the P08514 /IIIa receptor , is an important biological therapy in the management of patients presenting with ACS . AREAS COVERED : The objective of this review is to define the role of abciximab in the management of ACS by interpreting the available data from randomized clinical trials using abciximab in various clinical scenarios , particularly in percutaneous coronary intervention ( P05154 ) . We also review different modes of delivery and describe the adverse effects of abciximab including thrombocytopenia . Where possible , we attempt to compare abciximab to the other available P08514 /IIIa inhibitors . We hope the reader will gain a better understanding of the benefits and risks of abciximab and the important role it has in the management of cardiology patients . EXPERT OPINION : DB00054 was a breakthrough drug in the management of high risk ACS patients undergoing P05154 . However , with newer available therapies and improvement in P05154 technology , dose and delivery of this drug have evolved as we try to extract maximum benefit while minimizing the adverse effects associated with it . Inhibitors of P11274 signalling interrupt the survival signal mediated by the micro-environment in mantle cell lymphoma . Several studies provide evidences for mantle cell lymphoma ( Q8WXI8 ) cell survival relying on B-cell receptor ( P11274 ) -mediated signalling pathways , whereas the nature of this activation is unknown . Significant progress in Q8WXI8 treatment is achieved through therapies targeting P11274 -associated kinases , i.e. , DB09053 and Fostamatinib , inhibitors of Q06187 and P43405 , respectively . Our study addresses survival signals emanating from the P11274 or the tumour environment and how inhibiting P11274 signalling effectors might impact these survival signals . We found that Q06187 was constitutively activated and that P43405 phosphorylation was highly increased and sustained upon P11274 activation of primary Q8WXI8 cells . Moreover , Q8WXI8 cells from leukaemic patients secreted high amount of IL-1β , P05231 , P10145 and P13501 . Activation of the P11274 induced ( i ) cell survival , ( ii ) P40763 activation and ( iii ) increased autocrine secretion of IL-1β , P05231 , P10145 , P13501 , P22301 , TNFα and P15692 . Specific inhibition of Q06187 by DB09053 or P43405 by Fostamatinib ( R406 ) reversed these protective effects and decreased both basal and P11274 -induced autocrine cytokine secretions associated with P40763 phosphorylation . Interestingly , targeting Q06187 and P43405 prevented and inhibited P11274 -induced Q8WXI8 cell adhesion to human bone marrow stromal cells ( HMSCs ) in short- and long-term co-culture . We demonstrated that P11274 -induced survival relies on autocrine secretion of IL-1β , TNFα and P13501 that might facilitate adhesion of Q8WXI8 cells to HMSC . Treatment with DB09053 or Fostamatinib blocked the chemotactic signal thus increasing apoptosis . Reduced satiating effect of d-fenfluramine in serotonin 5-HT(2C) receptor mutant mice . RATIONALE : d- DB00574 stimulates the release of serotonin ( 5-HT ) and is a potent inhibitor of the re-uptake of 5-HT into nerve terminals . Administration of d-fenfluramine suppresses food intake in both animals and humans . OBJECTIVE : We have investigated the role of the P28335 receptor in mediating the effect of d-fenfluramine on mouse food intake and the behavioural satiety sequence . METHODS : Mutant mice lacking serotonin P28335 receptors and wild-type animals were habituated to a daily presentation of wet mash . Animals were non-deprived and received d-fenfluramine ( 3-30 mg/kg ) 30 min prior to being assessed for the presence of stereotypy and presented with wet mash . The behaviour of animals was observed for the subsequent 40 min and food intake was recorded . RESULTS : d- DB00574 dose-dependently inhibited the consumption of a palatable wet mash by the mice . d- DB00574 ( 3 mg/kg ) significantly reduced the amount of wet mash consumed by wild-type mice and induced a temporal advance in the behavioural satiety sequence consistent with an enhancement of satiety . Mutant mice were less sensitive to the satiating effects of 3 mg/kg d-fenfluramine . Hence , this dose of d-fenfluramine had a reduced effect on both food consumption and the behavioural satiety sequence in the P28335 mutant mice . In contrast , mutant mice showed an increased sensitivity to the stereotypy induced by high doses of d-fenfluramine ( 10 , 30 mg/kg ) compared to that of wild-type littermates . CONCLUSION : These data demonstrate a role for the P28335 receptor in mediating d-fenfluramine-induced satiety .

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interacts_with DB00009?

multiple_choice

Hereditary angioedema : a Taiwanese family with a novel gene mutation . Hereditary angioedema ( HAE ) is an autosomal dominant disorder caused by a deficiency of P05155 ( DB05341 ) . Affected individuals have attacks of swelling involving almost any part of the body . We studied a family with 15 living members , including a 16-year-old girl who had 3 attacks of angioedema in 2 years . Her paternal uncle had died of asphyxiation during an attack 15 years previously . We analyzed the blood of each family member for P01024 , C4 , and DB05341 levels and sequenced the P05155 ( formerly P05155 ) gene that codes for DB05341 . Six individuals had decreased serum levels of C4 and DB05341 , and they were all found to have a single nucleotide A deletion at codon 210 of the gene , 1210fsX210 , a novel mutation that accounts for the HAE in this family . Internalization of OspA in rsCD14 complex and aggregated forms . Although the spirochetal protein OspA is capable of stimulating immune cells in a P08571 - and O60603 -dependent manner , little is known about how O60603 receptor complex ligands , such as OspA , are handled by the cell once delivered . We examine here the internalization of the fluorescently derivatized forms of both the full length DB00045 delivered as a recombinant soluble P08571 ( rsCD14 ) complex and the corresponding lipohexapeptide given to the cells as an aggregate . Both forms of OspA are internalized in a similar manner to acetylated low density lipoprotein ( AcLDL ) , a scavenger receptor ligand . Acetylated low density lipoprotein is capable of competing for internalization with OspA even when OspA is delivered as a rsCD14 complex . We observe co-localization of OspA with lysosomes but not with the Golgi complex . These phenomena are similar between RAW264.7 macrophages and endothelial cells but change drastically when the cells are deprived of serum . Upon serum starvation , OspA shows some localization to the Golgi apparatus whereas the lipohexapeptide remains on the cell surface . Inhibition of internalization of OspA via treatment with cytochalasin D or of the lipohexapeptide via serum starvation does not interfere with P01375 induction activity , consistent with signalling from the cell surface . The molecular mechanism of osteoclastogenesis in rheumatoid arthritis . Bone-resorbing osteoclasts are formed from hemopoietic cells of the monocyte-macrophage lineage under the control of bone-forming osteoblasts . We have cloned an osteoblast-derived factor essential for osteoclastogenesis , the receptor activator of NF-kappaB ligand ( O14788 ) . Synovial fibroblasts and activated T lymphocytes from patients with rheumatoid arthritis also express O14788 , which appears to trigger bone destruction in rheumatoid arthritis as well . Recent studies have shown that T lymphocytes produce cytokines other than O14788 such as Q16552 , granulocyte-macrophage colony-stimulating factor and P01579 , which have powerful regulatory effects on osteoclastogenesis . The possible roles of O14788 and other cytokines produced by T lymphocytes in bone destruction are described . Identification of human thioredoxin as a novel P01579 -induced factor : mechanism of induction and its role in cytokine production . BACKGROUND : P01579 is a multifunctional peptide with a potent immune defense function which is also known as a prototypic Th1 cytokine . While screening for genes differentially expressed by Th1 and Th2 cytokines , human thioredoxin was identified as a novel target gene induced by P01579 . The mechanism by which thioredoxin is induced by P01579 and the signaling pathways involved in its induction were analyzed . In addition , the effects of thioredoxin on immune cell survival and cytokine production were examined by thioredoxin over-expression and recombinant thioredoxin treatment . RESULTS : Human thioredoxin was selectively induced by P01579 in monocytic and T cell lines . In monocytic cells , the induction of thioredoxin gene expression by P01579 was dose-dependent , and both the mRNA and protein levels were increased by 2~3 fold within 4 to 24 h hours of P01579 treatment . The thioredoxin induction by P01579 was insensitive to cycloheximide treatment , suggesting that it is a primary response gene induced by P01579 . Subsequent analysis of the signaling pathways indicated that the Jak/Stat , Akt , and Erk pathways play a role in P01579 signaling that leads to thioredoxin gene expression . P10599 was induced by oxidative or radiation stresses , and it protected the immune cells from apoptosis by reducing the levels of reactive oxygen species . Furthermore , thioredoxin modulated the oxidant-induced cytokine balance toward Th1 by counter-regulating the production of P05112 and P01579 in T cells . CONCLUSION : These data suggest that thioredoxin is an P01579 -induced factor that may play a role in developing Th1 immunity and in the maintenance of immune homeostasis upon infection , radiation , and oxidative stress . Upregulation of cell-surface-associated plasminogen activation in cultured keratinocytes by interleukin-1 beta and tumor necrosis factor-alpha . Keratinocytes synthesize and secrete urokinase-type plasminogen activator ( uPA ) which is bound in an autocrine manner to a specific receptor ( uPA-R ) at the keratinocyte surface . P00747 that is also bound to specific membrane binding sites is readily activated by uPA-R-bound uPA . Thus , plasmin is provided for proteolysis of pericellular glycoproteins . The expression of uPA and the uPA-R is confined to migrating keratinocytes during epidermal wound healing , rather than to keratinocytes of the normal epidermis . The regulatory factors of uPA/uPA-R expression in keratinocytes remained largely elusive . Proinflammatory cytokines , such as tumor necrosis factor-alpha ( P01375 ) or interleukin-1 beta ( P01584 ) , are present in epidermal wounds . We have therefore tested P01584 and P01375 for their influence on surface-associated plasminogen activation in a human keratinocyte cell line ( HaCaT ) as well as in primary cultures of normal human epidermal keratinocytes . Both cytokines induced the secretion of uPA into the culture supernatants and a concomitant increase in uPA activity as well as in uPA and uPA-R antigen at the cell surface . The increase was preceded by an increase in specific mRNA . The induction was accompanied by an accelerated uPA-dependent and plasmin-mediated detachment of HaCaT cells from the culture substratum . Taken together , the proinflammatory cytokines P01584 and P01375 induced a coordinated increase in uPA and uPA-R as well as increased pericellular plasmin-mediated proteolysis in human epidermal keratinocytes . This function might be an element of the molecular cell biological events during epidermal wound healing . DB11582 suppresses osteoclastogenesis induced by O14788 and cancer cells through inhibition of inflammatory pathways : a new use for an old drug . BACKGROUND AND PURPOSE : Most patients with cancer die not because of the tumour in the primary site , but because it has spread to other sites . Common tumours , such as breast , multiple myeloma , and prostate tumours , frequently metastasize to the bone . To search for an inhibitor of cancer-induced bone loss , we investigated the effect of thiocolchicoside , a semi-synthetic colchicoside derived from the plant Gloriosa superba and clinically used as a muscle relaxant , on osteoclastogenesis induced by receptor activator of NF-κB ligand ( O14788 ) and tumour cells . EXPERIMENTAL APPROACH : We used RAW 264.7 ( murine macrophage ) cells , a well-established system for osteoclastogenesis , and evaluated the effect of thiocolchicoside on O14788 -induced NF-κB signalling and osteoclastogenesis as well as on osteoclastogenesis induced by tumour cells . KEY RESULTS : DB11582 suppressed osteoclastogenesis induced by O14788 , and by breast cancer and multiple myeloma cells . Inhibition of the NF-κB pathway was responsible for this effect since the colchicoside inhibited O14788 -induced NF-κB activation , activation of IκB kinase ( IKK ) and suppressed inhibitor of NF-κBα ( IκBα ) phosphorylation and degradation , an inhibitor of NF-κB . Furthermore , an inhibitor of the IκBα kinase γ or NF-κB essential modulator , the regulatory component of the IKK complex , demonstrated that the NF-κB signalling pathway is mandatory for osteoclastogenesis induced by O14788 . CONCLUSIONS AND IMPLICATIONS : Together , these data suggest that thiocolchicoside significantly suppressed osteoclastogenesis induced by O14788 and tumour cells via the NF-κB signalling pathway . Thus , thiocolchicoside , a drug that has been used for almost half a century to treat muscle pain , may also be considered as a new treatment for bone loss . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . Novel mutations in the 1alpha-hydroxylase ( P450c1 ) gene in three families with pseudovitamin D-deficiency rickets resulting in loss of functional enzyme activity in blood-derived macrophages . Pseudovitamin D-defiency rickets ( PDDR ) is an autosomal recessive disorder characterized by hypocalcemia , rickets ( which are resistant to treatment with vitamin D ) , and low or undetectable serum levels of 1,25-dihydroxyvitamin D ( 1,25(OH)2D ) . The symptoms are corrected with 1,25(OH)2D treatment , and the disease is now believed to result from a defect in the cytochrome P450 component ( P450c1 ; O15528 ) of the renal 25-hydroxyvitamin D-1alpha-hydroxylase ( 1-OHase ) . We have studied genomic DNA from three families with PDDR and have identified the same homozygous mutation in the P450c1 gene in two of the index cases , causing a frameshift in exon 8 , resulting in a premature stop codon in the heme-binding domain . The two cases in the third kindred were compound heterozygotes with missense mutations in exons 6 and 9 . We have also identified a C/T polymorphism in intron 6 of the P450c1 genomic DNA . P01579 -inducible 1-OHase activity in blood-derived macrophages was shown by 1,25(OH)2D synthesis in all control cells tested ( 37-184 fmol/h/106 cells ) and those from the PDDR family parents ( 34-116 fmol/h/106 cells ) but was totally absent from the patients ' cells , indicating a defect in their macrophage 1-OHase , similar to the presumed renal defect . The assumption of similarity between the renal and macrophage P450c1 was supported by our ability to clone a 514 bp sequence , including the heme-binding region of the macrophage P450c1 cDNA from controls , which was identical to that published for both the renal and keratinocyte P450c1 cDNAs . Successful thrombolysis of a stroke with a pulmonary embolism in a young woman . BACKGROUND : Paradoxical embolism is a rare event , accounting for < 2 % of all arterial emboli . The diagnosis is often difficult , and consequences for the patient can be severe . CASE REPORT : We describe the case of a 35-year-old female physician who presented to our Emergency Department ( ED ) in severe hemodynamic compromise , with an altered level of consciousness and major expressive aphasia 1 day after undergoing a leg varicosal stripping procedure under regional anesthesia . She was successfully thrombolyzed with 0.9 mg/kg of Recombinant Tissue P00747 Activator ( rtPA , DB00009 ) and had a full recovery . CONCLUSION : To our knowledge , this is the first description of a case of massive pulmonary embolism associated with a paradoxical stroke related to patent foramen ovale that was thrombolyzed for both conditions with a " neurological dose " of rtPA . Although thrombolysis was completely successful in this case , indications and contraindications should be thoroughly respected . A more conservative approach with anticoagulation , or a more aggressive approach with surgical thrombectomy , can each potentially have a place in particular cases . Intra-arterial catheter-directed thrombolysis and percutaneous embolectomy are additional options to be considered when available , especially if there are contraindications for systemic thrombolysis . Q9H244 receptors play a significant role in the development of platelet microaggregation in patients with diabetes . Ninety-eight diabetic patients ( type 2 ) were studied together with 24 healthy normotensive controls . Microaggregates ( particle scale , < 25 microm ) of platelets were detected by a laser scattering system . Microaggregates in the control group showed a time-dependent reversible change ; however , they existed continuously in 82 of 98 diabetic patients . When platelets of diabetics were stimulated by a shear stress alone without ADP , 74 also showed spontaneous and irreversible microaggregates even though they were not observed in all control subjects . In control subjects , microaggregates were inhibited by MRS2279 ( a P47900 antagonist ) , but not AR-C69931MX ( a Q9H244 antagonist ) . However , AR-C69931MX prevented irreversible microaggregates in diabetic patients . When either aspirin or ticlopidine was administered to diabetic patients with irreversible microaggregates , both drugs significantly decreased microaggregates induced by a low dose of ADP . DB00208 additionally reduced the microaggregates induced by shear stress alone . In conclusion , microaggregates of platelets via Q9H244 receptors could play a key role in the hypersensitivity of platelets in diabetic patients , and the measurement of microaggregation could be a useful marker to estimate of thrombogenesis . These findings present a possible new means for patients with diabetes to prevent ischemic events . Synthetic delivery system for tuberculosis vaccines : immunological evaluation of the M. tuberculosis 38 kDa protein entrapped in biodegradable P00747 microparticles . Tuberculosis remains a major public health burden which could be ameliorated by effective and well-defined subunit vaccines , particularly because the protective efficacy of current M. bovis BCG vaccines is both unpredictable and variable . The immunodominant 38 kDa antigen from Mycobacterium tuberculosis was entrapped in biodegradable poly ( DL-lactide co-glycolide ) ( P00747 ) microparticles which served as a delivery system . Both cellular and humoral immune responses were assessed and compared with those obtained after immunization with the 38 kDa protein emulsified in incomplete Freund 's adjuvant ( IFA ) . Vaccination of mice with a single dose of antigen-loaded microparticles resulted in specific IgG titres peaking after five weeks comparable to those achieved after vaccination with protein emulsified in incomplete Freund 's adjuvant ( IFA ) . T-cell responses were found to be superior to those induced with antigen/IFA . The T- and B-cell epitope specificities ad judged with synthetic peptides were identical following immunization with antigen in microparticles or IFA . Differences in adjuvanticity were revealed by measuring antigen-specific IgG1 , IgG2a and antigen-induced P01579 secretion in vitro : substantially higher titres of IgG2a were observed following immunization with antigen/microparticles than with 38 kDa protein/IFA . This was paralleled by a tenfold higher secretion of P01579 in mice injected with antigen/microparticles . Reduction in colony-forming units was not consistent in mice immunized with 38 kDa protein entrapped in microparticles which were subsequently infected with live tubercle bacilli . Taken together these results indicate that biodegradable P00747 microparticles constitute a favorable candidate vaccine delivery system worthy of further assessment in the quest to develop better and defined agents protecting against tuberculosis . DB01093 -treated HL60 cells : a model of neutrophil-like cells mainly expressing Q07343 subtype . The human promyelocytic HL60 cells acquired a neutrophilic phenotype after a 7- to 10-day DB01093 treatment . Fc gammaRII was up-regulated . Fc gammaRI was also up-regulated by an additional P01579 treatment . These cells are able to produce O2*- by NADPH oxidase activation in the presence of immune complexes or phorbol-12-myristate-13-acetate ( PMA ) . A change of their DB05876 subtype profile was also observed : Q07343 was the predominant isoenzyme , Q08499 was down-regulated and P27815 was no longer detectable . Additionally , the more NADPH oxidase was activated by PMA , the less P27815 was expressed , suggesting that NADPH oxidase activity could be used as a surrogate marker of P27815 down-regulation . DB01954 and DB03849 ( cilomilast ) , two selective DB05876 inhibitors , dose-dependently inhibited receptor-coupled activation of superoxide . These results suggest that Q07343 is the main subtype involved in regulating superoxide induced by Fc gammaRs activation . Furthermore , these cells , expressing almost exclusively Q07343 subtype , could be useful to identify selective Q07343 inhibitors . Glutamate modulators as potential therapeutic drugs in schizophrenia and affective disorders . Severe psychiatric disorders such as schizophrenia are related to cognitive and negative symptoms , which often are resistant to current treatment approaches . The glutamatergic system has been implicated in the pathophysiology of schizophrenia and affective disorders . A key component is the dysfunction of the glutamatergic N-methyl-D-aspartate ( DB01221 ) receptor . Substances regulating activation/inhibition of the DB01221 receptor have been investigated in schizophrenia and major depression and are promising in therapeutic approaches of negative symptoms , cognition , and mood . In schizophrenia , add-on treatments with glycine , D-serine , D-alanine , D-cycloserine , D-amino acid oxidase inhibitors , glycine transporter-1 ( P48067 ) inhibitors ( e.g. , sarcosine , bitopertin ) and agonists ( e.g. , DB05096 ) or positive allosteric modulator ( e.g. , ADX71149 ) of group II metabotropic glutamate receptors ( mGluRs ) have been studied . In major depression , the DB01221 receptor antagonists ( e.g. , ketamine , AZD6765 ) , Q13224 subtype antagonists ( e.g. , traxoprodil , MK-0657 ) , and partial agonists ( e.g. , D-cycloserine , GLYX-13 ) at the glycine site of the DB01221 receptor have been proven to be effective in animal studies and first clinical trials . In addition , clinical studies of Q14416 /3 antagonist BCI-838 ( a prodrug of BCI-632 ( MGS0039 ) ) , Q14416 /3-negative allosteric modulators ( NMAs ) ( e.g. , RO499819 , RO4432717 ) , and P41594 NAMs ( e.g. , AZD2066 , RO4917523 ) are in progress . Future investigations should include effects on brain structure and activation to elucidate neural mechanisms underlying efficacy of these drugs . Stimulation of purinergic receptors modulates chemokine expression in human keratinocytes . DB00171 is abundantly released from stressed or damaged cells in response to mechanical stimulation , bacteria , or noxious agents . In this study , we have investigated the possible involvement of P2 receptors ( receptor for extracellular nucleotides ) in the expression and release of inflammatory mediators by human keratinocytes . Notably , extracellular DB00171 displayed a complex regulation of P01579 -stimulated chemokine expression , with upregulation of CC chemokine ligand 2 ( P13500 ) , P13501 and CXC chemokine ligand 8 ( P10145 ) , and suppression of the receptor CXC chemokine receptor 3 ( P49682 ) , Q07325 , P02778 , and O14625 . The effect of DB00171 was mimicked by ADP and adenosine-5'-O-3-thiotriphosphate , whereas 2',3'-O-(4-benzoylbenzoyl) DB00171 ( BzATP ) downmodulated all chemokines investigated . UTP had no effect on P01579 -stimulated chemokine secretion . The broad-spectrum P2 receptor antagonist suramin and the selective P47900 inhibitor adenosine 3'-phosphate 5'-phosphosulfate counteracted the effect of DB00171 on secretion of all the chemokines examined , whereas pyridoxal phosphate 6-azophenyl 2',4'-disulfonic acid and KN62 ( 1- [ N,O-bis ( 5-isoquinoline sulfonyl ) -N-methyl-L-tyrosyl ] 4 phenylpiperazine ) partially prevented the inhibitory effect of DB00171 on P02778 secretion , but on the other hand potentiated the DB00171 -stimulatory effect on P13501 , P13500 , and P10145 release . In lesional skin of psoriasis and atopic dermatitis patients , intense Q99572 reactivity was confined to the cell membrane of the basal layer , whereas diffuse P47900 immunostaining was found throughout the epidermis . Collectively , our data suggest that the orchestrated activation of distinct P2Y and P2X receptors modulates skin inflammation . Changes in the phenotype of polymorphic plasma proteins after liver transplantation - new data and medico-legal consequences . The genetically inherited polymorphic plasma protein types have always been considered stable for lifetime in humans . Most of these proteins are synthetised in the liver . Phenotypes for 14 plasma proteins in donors and recipients of liver transplants prior to and after transplantation were determined in 15 patients who had undergone liver transplantation at the university hospitals Charité and Rudolf Virchow in Berlin . The plasma proteins investigated were HP , TF , GC , PI , P02763 , ITI , A2HS , P00747 , FXIIIB , BF , P01024 , P13671 , Q99618 and FH . Evidence was provided of irreversible change from the recipient type to the donor type in at least one patient for all the systems investigated . This is the first time such data have been obtained for ITI , A2HS , Q99618 and FH . These results clearly support the point that the dogma of life-long stability of genetically determined protein phenotypes is merely of limited validity . Against the background of good long-term results of liver transplantation , there are consequences for the practice of legal medicine in the particular context of certification of parentage , identification and stain analysis . Vitamin D metabolism , mechanism of action , and clinical applications . DB00169 is made in the skin from 7-dehydrocholesterol under the influence of UV light . DB00153 ( ergocalciferol ) is derived from the plant sterol ergosterol . Vitamin D is metabolized first to 25 hydroxyvitamin D ( 25OHD ) , then to the hormonal form 1,25-dihydroxyvitamin D ( 1,25(OH)2D ) . Q6VVX0 is the most important 25-hydroxylase ; O15528 is the key 1-hydroxylase . Both 25OHD and 1,25(OH)2D are catabolized by Q07973 . 1,25(OH)2D is the ligand for the vitamin D receptor ( P11473 ) , a transcription factor , binding to sites in the DNA called vitamin D response elements ( VDREs ) . There are thousands of these binding sites regulating hundreds of genes in a cell-specific fashion . P11473 -regulated transcription is dependent on comodulators , the profile of which is also cell specific . Analogs of 1,25(OH)2D are being developed to target specific diseases with minimal side effects . This review will examine these different aspects of vitamin D metabolism , mechanism of action , and clinical application . DB01954 attenuates acute oligodendrocyte death in the adult rat ventrolateral funiculus following contusive cervical spinal cord injury . DB01954 , an inhibitor of phosphodiesterase 4 ( DB05876 ) proteins that hydrolyze DB02527 , increases axonal regeneration following spinal cord injury ( SCI ) . Recent evidence indicate that rolipram also protects against a multitude of apoptotic signals , many of which are implicated in secondary cell death post-SCI . In the present study , we used immunohistochemistry and morphometry to determine potential spinal cord targets of rolipram and to test its protective potential in rats undergoing cervical spinal cord contusive injury . We found that 3 DB05876 subtypes ( P27815 , B , D ) were expressed by spinal cord oligodendrocytes . OX-42 immunopositive microglia only expressed the Q07343 subtype . Oligodendrocyte somata were quantified within the cervical ventrolateral funiculus , a white matter region critical for locomotion , at varying time points after SCI in rats receiving rolipram or vehicle treatments . We show that rolipram significantly attenuated oligodendrocyte death at 24 h post-SCI continuing through 72 h , the longest time point examined . These results demonstrate for the first time that spinal cord glial cells express DB05876 subtypes and that the DB05876 inhibitor rolipram protects oligodendrocytes from secondary cell death following contusive SCI . They also indicate that further investigations into neuroprotection and axonal regeneration with rolipram are warranted for treating SCI . Relationship between the antidiarrhoeal effects of Hange-Shashin-To and its active components . This study was designed to examine the relationship between the antidiarrhoeal effects of Hange-Shashin-To ( TJ-14 ) and its active components . Oral treatment with TJ-14 at 1000 mg/kg significantly inhibited castor oil-induced diarrhoea . Both the 50 % methanol eluate fraction ( fraction III ) and the methanol eluate fraction ( fraction IV ) showed antidiarrhoeal effects at oral doses of 68 mg/kg and 63 mg/kg , respectively , corresponding to 1000 mg/kg of TJ-14 . TJ-14 ( 1000 mg/kg , p.o. ) showed a significant increase in blood corticosterone levels . Increased blood corticosterone was noted after the oral administration of 63 mg/kg of fraction IV . The inhibitory activity of TJ-14 on cyclooxygenase-2 ( P35354 ) was also observed in fractions III and IV . The main component of fraction III was Scutellariae Radix-derived baicalin . Fraction IV contained Glycyrrhizae Radix-derived glycyrrhizin and isoliquiritin , Coptidis Rhizoma-derived berberine , coptisine and palmitine . DB01404 Radix-derived saponins were also present in fraction IV . These compounds inhibited castor-oil induced diarrhoea at oral doses of 10 or 30 mg/kg . Thus , the present results indicate that Scutellariae Radix , Glycyrrhizae Radix , DB01404 radix and Coptidis Rhizoma-derived components are involved in the antidiarrhoeal action of TJ-14 . 17beta-estradiol activates DB00947 -sensitive estrogen receptors and cyclic GMP-dependent thioredoxin expression for neuroprotection . Clinical studies suggest that estrogen may improve cognition in Alzheimer 's patients . Basic experiments demonstrate that 17beta-estradiol protects against neurodegeneration in both cell and animal models . In the present study , a human SH-SY5Y cell model was used to investigate molecular mechanisms underlying the receptor-mediated neuroprotection of physiological concentrations of 17beta-estradiol . 17beta-estradiol ( < 10 nM ) concomitantly increased neuronal nitric oxide synthase ( NOS1 ) expression and cell viability . 17beta-estradiol-induced neuroprotection was blocked by the receptor antagonist DB00947 , also prevented by inhibitors of NOS1 ( 7-nitroindazole ) , guanylyl cyclase ( LY 83,583 ) , and cGMP-dependent protein kinase ( PKG ) ( Rp-8-pCPT-cGMPs ) . In addition to the expression of NOS1 and MnSOD , 17beta-estradiol increased the expression of the redox protein thioredoxin ( P10599 ) , which was blocked by the inhibition of either cGMP formation or PKG activity . The expression of heme oxygenase 2 and brain-derived neurotrophic factor was not altered . P03372 -enhanced cell viability against oxidative stress may be linked to P10599 expression because the P10599 reductase inhibitor , 5,5'-dithio-bis ( 2-nitrobenzoic acid ) significantly reduced the cytoprotective effect of 17beta-estradiol . Furthermore , P10599 ( 1 microM ) inhibited lipid peroxidation , proapoptotic caspase-3 , and cell death during oxidative stress caused by serum deprivation . We conclude that cGMP-dependent expression of P10599 -- the redox protein with potent antioxidative and antiapoptotic properties -- may play a pivotal role in estrogen-induced neuroprotection . The antifibrotic effects of plasminogen activation occur via prostaglandin E2 synthesis in humans and mice . P00747 activation to plasmin protects from lung fibrosis , but the mechanism underlying this antifibrotic effect remains unclear . We found that mice lacking plasminogen activation inhibitor-1 ( P05121 ) , which are protected from bleomycin-induced pulmonary fibrosis , exhibit lung overproduction of the antifibrotic lipid mediator prostaglandin E2 ( DB00917 ) . P00747 activation upregulated DB00917 synthesis in alveolar epithelial cells , lung fibroblasts , and lung fibrocytes from saline- and bleomycin-treated mice , as well as in normal fetal and adult primary human lung fibroblasts . This response was exaggerated in cells from Pai1-/- mice . Although enhanced DB00917 formation required the generation of plasmin , it was independent of proteinase-activated receptor 1 ( P25116 ) and instead reflected proteolytic activation and release of P14210 with subsequent induction of P35354 . That the P14210 / P35354 / DB00917 axis mediates in vivo protection from fibrosis in Pai1-/- mice was demonstrated by experiments showing that a selective inhibitor of the P08581 c- DB00134 increased lung collagen to WT levels while reducing P35354 protein and DB00917 levels . Of clinical interest , fibroblasts from patients with idiopathic pulmonary fibrosis were found to be defective in their ability to induce P35354 and , therefore , unable to upregulate DB00917 synthesis in response to plasmin or P14210 . These studies demonstrate crosstalk between plasminogen activation and DB00917 generation in the lung and provide a mechanism for the well-known antifibrotic actions of the fibrinolytic pathway . Activation of spinal alpha-7 nicotinic acetylcholine receptor attenuates remifentanil-induced postoperative hyperalgesia . The activation of alpha-7 nicotinic acetylcholine receptors ( α7-nAchRs ) are currently being considered as novel therapeutic approaches for managing hyperalgesia in inflammation and chronic neuropathic pain , but the role of a7-nAChRs on opioids induced hyperalgesia remain unknown . The present study investigated the effects of α7-nAChRs selective agonists PHA-543613 and type II positive allosteric modulators ( PAMs ) PNU-120596 in remifentanil induced postoperative hyperalgesia . As the results shown , intrathecal treatment with both α7-nAChRs agonists and type II PAMs could attenuate remifentanil induced hyperalgesia by increasing paw withdrawal mechanical threshold ( PWMT ) and paw withdrawal thermal latency ( PWTL ) . Furthermore , we also investigated the protein level of proinflammatory cytokines and phosphorylation N-methyl-d-aspartate receptor 2B subunit ( p- Q13224 ) in the spinal cord . Our data indicated that activation of α7-nAchRs decreased the proinflammatory cytokines ( P01375 -α , P05231 ) and p- Q13224 protein level in the spinal cord . The depression of the increased levels of proinflammatory cytokines and p- Q13224 after remifentanil treatment may contribute to the anti-hyperalgesia effects of PHA-543613and PNU-120596 via α7-nAChRs . Therefore , our findings demonstrated that α7-nAChRs may be potential candidates for treating opioids induced hyperalgesia .

MH_train_1339

MH_train_1339

MH_train_1339

interacts_with DB00215?

multiple_choice

Serotonergic mechanisms in human allergic contact dermatitis . Expression of serotonin ( 5-hydroxytryptamine ; 5-HT ) , 5-HT receptors 1A ( 5-HT1AR ) and 2A , and serotonin transporter protein ( P31645 ) was studied in positive epicutaneous reactions to nickel sulphate in nickel-allergic patients , at 72 h post-challenge with the antigen . In addition , the effects of 5-HT2AR agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , and the selective serotonin reuptake inhibitors ( SSRIs ) citalopram and fluoxetine , were tested on nickel-stimulated peripheral blood mononuclear cells from nickel-allergic patients , regarding their proliferation and interleukin ( IL ) -2 production , as well as the effect of these SSRIs on a murine Langerhans ' cell-like line ( XS52 ) , regarding its IL-1beta production . Serotonin-positive platelets were increased in the inflamed skin compared with control skin . A decrease ( p < 0.01 ) in 5-HT1AR-positive mononuclear cells was evident in the eczematous skin compared with control skin , whereas 5-HT2AR- and P31645 -positive cells were increased ( p < 0.001 for both ) in the eczematous skin . Treatment of nickel-stimulated peripheral blood mononuclear cells with 5x10(-5) mol/l of DOI inhibited ( p < 0.01 ) the proliferation of nickel-stimulated peripheral blood mononuclear cells , while no effect was found regarding P60568 production . DB00215 at 10(-6) mol/l tended to inhibit the production of IL-1beta by the XS52 cell line . These results indicate the implication of the serotonergic system in the contact allergic reaction . P43490 / P43490 /visfatin and cancer . P43490 / P43490 /visfatin is the rate-limiting enzyme that catalyzes the first step in NAD biosynthesis from nicotinamide and regulates growth , apoptosis and angiogenesis of mammalian cells . This enzyme was originally cloned as a putative cytokine shown to enhance the B cell precursor maturation in the presence of P13232 and stem cell factor . A number of cancers have increased expression of P43490 / P43490 /visfatin , which regulates a variety of different signaling pathways such as PI3K/Akt , P27361 /2 and P40763 . FK866/APO866 and CHS828/ DB05217 are two known inhibitors of P43490 / P43490 /visfatin and have been evaluated as anticancer agents in the clinic . This review will focus on its role in carcinogenesis and cancer progression and its inhibitors as therapeutic target for cancer treatment . Computation of standard binding free energies of polar and charged ligands to the glutamate receptor P42262 . Accurate calculation of the binding affinity of small molecules to proteins has the potential to become an important tool in rational drug design . In this study , we use the free energy perturbation ( FEP ) method with restraints to calculate the standard binding free energy of five ligands ( ACPA , AMPA , CNQX , DB03759 , and glutamate ) to the glutamate receptor P42262 , which plays an essential role in synaptic transmission . To deal with the convergence problem in FEP calculations with charged ligands , we use a protocol where the ligand is coupled in the binding site while it is decoupled in bulk solution simultaneously . The contributions from the conformational , rotational , and translational entropies to the standard binding free energy are determined by applying/releasing respective restraints to the ligand in bulk/binding site . We also employ the confine-and-release approach , which helps to resolve convergence problems in FEP calculations . Our results are in good agreement with the experimental values for all five ligands , including the charged ones which are often problematic in FEP calculations . We also analyze the different contributions to the binding free energy of each ligand to P42262 and discuss the nature of these interactions . NF-kappaB inhibitory action of resveratrol : a probable mechanism of neuroprotection in experimental diabetic neuropathy . DB02709 has shown array of biological actions , and is under clinical development for various disease conditions . The etiology of diabetic neuropathy revolves around oxidative stress , P51606 formation , lipid peroxidation etc . All these stimulate inflammatory processes and NF-kappaB cascade is considered as one of the major players of inflammatory response . Activation of NF-kappaB results in elevated levels of inflammatory mediators . P35354 and P01375 activity have also been correlated with inflammatory damage in the pathophysiology of diabetic neuropathy ( DN ) . Therefore we investigated the effect of resveratrol on NF-kappaB inflammatory cascade , P35354 , P01375 and P05231 levels in experimental DN . We found that resveratrol protected against various functional and behavioral deficits in diabetic neuropathy in line with our earlier published reports . In this study we found that the resveratrol treatment decreased the expression of p65 and IkappaB-alpha in treated rats . Treatment also ameliorated the elevated levels of P01375 , P05231 and P35354 . DB02709 treatment produced significant decrease in nerve MDA levels in treated animals which may also be contributing to reduction in neuro-inflammation . This study confirms the NF-kappaB inhibitory activity and anti-inflammatory activity of resveratrol which may contribute to neuroprotection in diabetic neuropathy apart from its antioxidant effect . P01375 alpha mediates GABA(A) receptor trafficking to the plasma membrane of spinal cord neurons in vivo . The proinflammatory cytokine TNFα contributes to cell death in central nervous system ( CNS ) disorders by altering synaptic neurotransmission . TNFα contributes to excitotoxicity by increasing P42262 -lacking AMPA receptor ( AMPAR ) trafficking to the neuronal plasma membrane . In vitro , increased AMPAR on the neuronal surface after TNFα exposure is associated with a rapid internalization of GABA(A) receptors ( GABA(A)Rs ) , suggesting complex timing and dose dependency of the CNS 's response to TNFα . However , the effect of TNFα on GABA(A)R trafficking in vivo remains unclear . We assessed the effect of TNFα nanoinjection on rapid GABA(A)R changes in rats ( N = 30 ) using subcellular fractionation , quantitative western blotting , and confocal microscopy . GABA(A)R protein levels in membrane fractions of TNFα and vehicle-treated subjects were not significantly different by Western Blot , yet high-resolution quantitative confocal imaging revealed that TNFα induces GABA(A)R trafficking to synapses in a dose-dependent manner by 60 min . TNFα-mediated GABA(A)R trafficking represents a novel target for CNS excitotoxicity . Novel treatments of GERD : focus on the lower esophageal sphincter . Up to 50 % of patients with gastroesophageal reflux disease ( GERD ) still suffer from GERD symptoms despite proton pump inhibitor ( PPI ) therapy , indicating a need for new treatments . The lower esophageal sphincter ( LES ) plays a crucial role in maintaining the mechanical barrier necessary for prevention of gastric reflux . Transient LES relaxation ( TLESR ) is an important factor behind the occurrence of reflux , and preclinical studies have identified a number of targets for pharmacologic modification of TLESR . However , only gamma-aminobutyric acid ( GABA ) type B receptor ( GABA(B) ) agonists and metabotropic glutamate receptor 5 ( P41594 ) modulators have shown positive proof of concept in the clinical setting . The P41594 negative allosteric modulator DB05070 improved symptoms in GERD patients , but was associated with central side effects such as dizziness . DB00181 , a GABA(B) receptor agonist , reduces the incidence of TLESR and improves GERD symptoms in both adult and pediatric GERD patients . However , the utility of baclofen is similarly limited by poor tolerability and recent research has focused on the development of GABA(B) receptor agonists with improved tolerability . DB05031 , a prodrug of R-baclofen , reduced the number of reflux episodes in a dose-ranging study and was similarly tolerated to placebo . AZD3355 and AZD9343 are GABA(B) receptor agonists with limited central nervous system activity that have been shown in preclinical studies to reduce the incidence of TLESR and decrease esophageal acid exposure ; data from clinical studies of these agents in GERD patients are awaited with interest . Agents that target TLESR activity may therefore offer a promising new add-on treatment for patients who suffer from GERD symptoms despite PPI therapy . Genetic variants contribute to gene expression variability in humans . Expression quantitative trait loci ( eQTL ) studies have established convincing relationships between genetic variants and gene expression . Most of these studies focused on the mean of gene expression level , but not the variance of gene expression level ( i.e. , gene expression variability ) . In the present study , we systematically explore genome-wide association between genetic variants and gene expression variability in humans . We adapt the double generalized linear model ( dglm ) to simultaneously fit the means and the variances of gene expression among the three possible genotypes of a biallelic SNP . The genomic loci showing significant association between the variances of gene expression and the genotypes are termed expression variability QTL ( evQTL ) . Using a data set of gene expression in lymphoblastoid cell lines ( LCLs ) derived from 210 HapMap individuals , we identify cis-acting evQTL involving 218 distinct genes , among which 8 genes , Q08828 , P26232 , Q86T65 , Q96AC1 , P05231 , O00469 , Q9UNH6 , and O00300 , are cross-validated using an extra expression data set of the same LCLs . We also identify ∼300 trans-acting evQTL between > 13,000 common SNPs and 500 randomly selected representative genes . We employ two distinct scenarios , emphasizing single-SNP and multiple-SNP effects on expression variability , to explain the formation of evQTL . We argue that detecting evQTL may represent a novel method for effectively screening for genetic interactions , especially when the multiple-SNP influence on expression variability is implied . The implication of our results for revealing genetic mechanisms of gene expression variability is discussed . In vivo effects of a combined P28222 receptor/ P31645 antagonist in experimental pulmonary hypertension . AIMS : A mechanism for co-operation between the serotonin ( 5-hydroxytryptamine , 5-HT ) transporter and P28222 receptor in mediating pulmonary artery vasoconstriction and proliferation of pulmonary artery smooth muscle cells has been demonstrated in vitro . Here we determine , for the first time , the in vivo effects of a combined P28222 receptor/serotonin transporter antagonist ( LY393558 ) with respect to the development of pulmonary arterial hypertension ( PAH ) and its in vitro effects in human pulmonary artery smooth muscle cells ( hPASMCs ) derived from idiopathic PAH ( IPAH ) patients . METHODS AND RESULTS : We determined the effects of LY393558 as well as a selective serotonin transporter inhibitor , citalopram , on right ventricular pressure , right ventricular hypertrophy , and pulmonary vascular remodelling in wildtype mice and mice over-expressing serotonin transporter ( P31645 + mice ) before and after hypoxic exposure . We also compared their effectiveness at reversing PAH in P31645 + mice and hypoxic mice . Further , we examined the proliferative response to serotonin in IPAH hPASMCs . We also clarified the pharmacology of serotonin-induced vasoconstriction and P28222 receptor/serotonin transporter interactions in mouse isolated pulmonary artery . DB00215 had a moderate effect at preventing and reversing experimental PAH in vivo whereas LY393558 was more effective . LY393558 was more effective than citalopram at reversing serotonin-induced proliferation in IPAH hPASMCs . There is synergy between P28222 receptor and serotonin transporter inhibitors against serotonin-induced vasoconstriction in mouse pulmonary arteries . CONCLUSION : P28222 receptor and serotonin transporter inhibition are effective at preventing and reversing experimental PAH and serotonin-induced proliferation of PASMCs derived from IPAH patients . Targeting both the serotonin transporter and P28222 receptor may be a novel therapeutic approach to PAH . Sharing of the P60568 receptor gamma chain with the functional Q01113 complex . The third subunit , the so-called common gamma ( gamma c ) chain , of the P60568 receptor is shared among the receptors for P60568 , P05112 , P13232 and P40933 , and dysfunction of the gamma c chain is thought to cause X-linked severe combined immunodeficiency ( XSCID ) ascribed to impairment of early T cell development . However , cytokines linked to XSCID are as yet unidentified . A mAb specific for the gamma c chain , TUGm2 , profoundly inhibited cell proliferation in response to P15248 . Another mAb , TUGm3 , immunoprecipitated [125I] P15248 cross-linked with either the Q01113 or the gamma c chain . These results demonstrate that the gamma c chain is included in the functional receptor complex for P15248 , which was initially characterized as a T cell growth factor and is essential for P15248 -dependent growth signal transduction . Chronic methamphetamine exposure alters immune function in normal and retrovirus-infected mice . Methamphetamine ( MA ) abuse represents a growing problem in the USA with an increase of sudden death . To evaluate the immune function alterations due to chronic methamphetamine use , we examined C57BL/C mice with LP-BM5 retrovirus infection plus methamphetamine exposure . Mice were randomly assigned to the following groups : placebo , placebo retrovirus-infected , uninfected MA treated and retrovirus-infected MA treated . Placebo , MA-treated groups were intraperitoneally injected with saline , MA , respectively , with a gradually increasing dose from 15 to 40 mg/kg for 12 weeks ( 5 days/week ) . Con A- and LPS-induced mitogenesis of splenocytes , cytokine production by splenocytes culture and lipid peroxides in the liver were measured . Heart tissue histopathology was analyzed in all the groups with murine cytomegalovirus ( CMV ) superinfection . Our data showed that MA treatment significantly decreased production of P60568 and interferon gamma ( P01579 ) in uninfected mice but did not further suppress the reduced Th1 cytokines in retrovirus-infected mice . There were no significant effects on cytokines P05112 and P05231 . However , tumor necrosis factor ( P01375 ) was significantly increased in both uninfected and infected mice due to MA treatment . Lipid peroxides in liver were significantly increased both in uninfected and retrovirus-infected mice due to MA exposure . DB00163 levels in liver were significantly decreased in uninfected mice due to MA treatment . CMV superinfection greatly increased the cardiac lesions in retrovirus-infected mice while no significant histopathology changes were detected due to MA treatment . Our data suggest that MA has immunomodulation activity , suppressing Th1 cytokine production and enhancing some Th2 cytokine secretion , as well as increasing lipid peroxides in uninfected mice . The interaction between LP-BM5 and MA remains unclear . Aerosol vaccination with AERAS-402 elicits robust cellular immune responses in the lungs of rhesus macaques but fails to protect against high-dose Mycobacterium tuberculosis challenge . Development of a vaccine against pulmonary tuberculosis may require immunization strategies that induce a high frequency of Ag-specific P01730 and CD8 T cells in the lung . The nonhuman primate model is essential for testing such approaches because it has predictive value for how vaccines elicit responses in humans . In this study , we used an aerosol vaccination strategy to administer AERAS-402 , a replication-defective recombinant adenovirus ( rAd ) type 35 expressing Mycobacterium tuberculosis Ags Ag85A , Ag85B , and TB10.4 , in bacillus Calmette-Guérin ( BCG ) -primed or unprimed rhesus macaques . Immunization with BCG generated low purified protein derivative-specific P01730 T cell responses in blood and bronchoalveolar lavage . In contrast , aerosolized AERAS-402 alone or following BCG induced potent and stable Ag85A/b-specific P01730 and CD8 effector T cells in bronchoalveolar lavage that largely produced IFN-γ , as well as P01375 and P60568 . Such responses induced by BCG , AERAS-402 , or both failed to confer overall protection following challenge with 275 CFUs M. tuberculosis Erdman , although vaccine-induced responses associated with reduced pathology were observed in some animals . Anamnestic T cell responses to Ag85A/b were not detected in blood of immunized animals after challenge . Overall , our data suggest that a high M. tuberculosis challenge dose may be a critical factor in limiting vaccine efficacy in this model . However , the ability of aerosol rAd immunization to generate potent cellular immunity in the lung suggests that using different or more immunogens , alternative rAd serotypes with enhanced immunogenicity , and a physiological challenge dose may achieve protection against M. tuberculosis . Immunopotentiation of bovine respiratory disease virus vaccines by interleukin-1 beta and interleukin-2 . Three experiments , using 85 crossbred beef calves , were conducted to evaluate the adjuvanticity of single , multiple , and combined doses of recombinant bovine P01584 ( rBoIL-1 beta ) and recombinant bovine P60568 ( rBoIL-2 ) , with a modified-live bovine herpesvirus-1/parainfluenza-3 ( BHV-1/ P19957 ) virus vaccine and a killed bovine viral diarrhea ( BVD ) virus vaccine . Cytokines were administered intramuscularly at vaccination but at different injection sites . All cytokine treatments increased non-major histocompatibility complex ( MHC ) -restricted cytolytic capability of peripheral blood mononuclear cells ( PBMC ) against virus-infected target cells and serum neutralizing ( SN ) antibody titers to BHV-1 and BVD virus . Multiple , consecutive injections of rBoIL-2 generally showed the greatest adjuvant effect , and no additive effect was observed when rBoIL-1 beta and rBoIL-2 were administered together . In a challenge experiment , calves were vaccinated with a modified-live BHV-1/ P19957 vaccine and infected with BHV-1 on Day 21 . Cytokine-treated calves had higher SN antibody titers to BHV-1 than did the control calves at the time of challenge . Calves that were administered rBoIL-2 on 5 consecutive days shed less BHV-1 and had the highest SN antibody titer to BHV-1 ( Day 28 ) . These data suggest that rBoIL-1 beta and rBoIL-2 may be useful immunoadjuvants for bovine respiratory disease virus vaccines . Inhibition of Q16552 as a pharmacological approach for Q9UKU7 . Several experimental approaches have been utilized , in order to critically examine the roles of Q16552 family members in intestinal inflammation . These approaches have included : ( 1 ) the use of Q16552 and Q96PD4 -deficient mice , ( 2 ) specific antibodies directed against Q16552 , ( 3 ) an Q16552 vaccine , ( 4 ) methods to block the Q16552 receptor and ( 5 ) small-molecule inhibitors of Q16552 . Previous studies found somewhat conflicting results in preclinical models of Inflammatory Bowel Disease ( Q9UKU7 ) , using specific strains of Q16552 -deficient mice . This paper will review the preclinical results using various pharmacological approaches [ specific Q16552 antibodies , an Q16552 receptor fusion protein , IL-12/IL-23 p40 subunit and Q16552 vaccine approaches , as well as a small molecule inhibitor ( Vidofludimus ) ] to inhibit Q16552 in animal models of Q9UKU7 . Recent clinical results in patients with Q9UKU7 will also be discussed for DB09029 ( an Q16552 antibody ) , Brodalumab ( an Q16552 receptor antibody ) and two small-molecule drugs ( Vidofludimus and DB08895 ) , which inhibit Q16552 as part of their overall pharmacological profiles . This review paper will also discuss some pharmacological lessons learned from the preclinical and clinical studies with anti- Q16552 drugs , as related to drug pharmacodynamics , Q16552 receptor subtypes and other pertinent factors . Finally , future pharmacological approaches of interest will be discussed , such as : ( 1 ) Retinoic acid receptor-related orphan nuclear receptor gamma t ( Rorγt ) antagonists , ( 2 ) P10276 ( RARα ) antagonists , ( 3 ) Pim-1 kinase inhibitors and ( 4 ) Dual small-molecule inhibitors of NF-κB and P40763 , like synthetic triterpenoids . Rapid inhibition of vasoconstriction in renal afferent arterioles by aldosterone . DB04630 has been suggested to elicit vessel contraction via a nongenomic mechanism . We tested this proposal in microdissected , perfused rabbit renal afferent arterioles . DB04630 had no effect on internal diameter in concentrations from 10(-10) to 10(-5) mol/L , but aldosterone abolished the ability of 100 mmol/L DB00761 to induce vascular contraction . The inhibitory effect of aldosterone was observed from 1 pmol/L . The inhibitory effect was significant after 5 minutes and maximal after 20 minutes and was fully reversible . DB00970 ( 10(-6) mol/L ) prolonged the effect of aldosterone . The effect was abolished by the mineralocorticoid receptor antagonist spironolactone ( 10(-7) mol/L ) but not by the glucocorticoid receptor antagonist mifepristone ( 10(-6) mol/L ) . The K+-mediated increase of intracellular calcium concentration in afferent arterioles was not affected by aldosterone . P08235 was detected by reverse transcription-polymerase chain reaction and immunohistochemistry in rat renal vasculature and rabbit endothelial cells . Inhibition of phosphatidylinositol ( PI ) -3 kinase with LY 294002 ( 3x10(-6) mol/L ) restored sensitivity to K+ in the presence of aldosterone , and afferent arterioles were immunopositive for P19957 kinase subunit P42336 . Inhibition of NO formation by L-NAME ( 10(-4) mol/L ) or inhibition of soluble guanylyl cyclase with 1H-(1,2,4)Oxadiazolo[4,3-a]quinoxaline-1-one restored K+-induced vasoreactivity in the presence of aldosterone . Similar to aldosterone , the NO donor sodium nitroprusside inhibited K+-induced vascular contraction . DB02424 ( 10(-6) mol/L ) , an inhibitor of heat shock protein 90 , abolished aldosterone-induced vasorelaxation . We conclude that aldosterone inhibits depolarization-induced vasoconstriction in renal afferent arterioles by a rapid nongenomic mechanism that is initiated by mineralocorticoid receptor activation and involves P19957 kinase , protein kinase B , and heat shock protein 90-mediated stimulation of NO generation . Analysis of a 26-kb region linked to the Mhc in zebrafish : genomic organization of the proteasome component beta/transporter associated with antigen processing-2 gene cluster and identification of five new proteasome beta subunit genes . Sequencing of zebrafish ( Danio rerio ) bacterial artificial chromosome and P1 artificial chromosome genomic clone fragments and of cDNA clones has led to the identification of five new loci coding for beta subunits of proteasomes ( PSMB ) . Together with the four genes identified previously , nine PSMB genes have now been defined in the zebrafish . Six of the nine genes reside in the zebrafish MHC ( Mhc ) class I region , four of them reside in a single cluster closely associated with TAP2 on a 26-kb long genomic fragment , and two reside at some distance from the fragment . In addition to homologues of the human genes P28074 through P28065 , two new genes , A5LHX3 and PSMB12 , have been found for which there are no known corresponding genes in humans . The new genes reside in the PSMB cluster in the Mhc . Homology and promoter region analysis suggest that the Mhc-associated genes might be inducible by P01579 . The zebrafish class I region contains representatives of three phylogenetically distinguishable groups of PSMB genes , X , Y , and Z . It is proposed that these genes were present in the ancestral PSMB region before Mhc class I genes became associated with it . DB00131 cyclase 1 ( Q08828 ) mutations cause recessive hearing impairment in humans and defects in hair cell function and hearing in zebrafish . DB02527 ( DB02527 ) production , which is important for mechanotransduction within the inner ear , is catalyzed by adenylate cyclases ( AC ) . However , knowledge of the role of ACs in hearing is limited . Previously , a novel autosomal recessive non-syndromic hearing impairment locus DFNB44 was mapped to chromosome 7p14.1-q11.22 in a consanguineous family from Pakistan . Through whole-exome sequencing of DNA samples from hearing-impaired family members , a nonsense mutation c.3112C > T ( p.Arg1038* ) within adenylate cyclase 1 ( Q08828 ) was identified . This stop-gained mutation segregated with hearing impairment within the family and was not identified in ethnically matched controls or within variant databases . This mutation is predicted to cause the loss of 82 amino acids from the carboxyl tail , including highly conserved residues within the catalytic domain , plus a calmodulin-stimulation defect , both of which are expected to decrease enzymatic efficiency . Individuals who are homozygous for this mutation had symmetric , mild-to-moderate mixed hearing impairment . Zebrafish adcy1b morphants had no FM1-43 dye uptake and lacked startle response , indicating hair cell dysfunction and gross hearing impairment . In the mouse , Adcy1 expression was observed throughout inner ear development and maturation . Q08828 was localized to the cytoplasm of supporting cells and hair cells of the cochlea and vestibule and also to cochlear hair cell nuclei and stereocilia . Ex vivo studies in COS-7 cells suggest that the carboxyl tail of Q08828 is essential for localization to actin-based microvilli . These results demonstrate that Q08828 has an evolutionarily conserved role in hearing and that DB02527 signaling is important to hair cell function within the inner ear . Randomised clinical trial : herbal extract DB05767 in active ulcerative colitis - a double-blind comparison with sustained release mesalazine . BACKGROUND : Andrographis paniculata is an herbal mixture used to treat inflammatory diseases . An extract of the herb , DB05767 , inhibits P01375 -α and IL-1β , and prevents colitis in animal models . AIM : To determine the efficacy and safety of DB05767 in patients with mild-to-moderate ulcerative colitis . METHODS : A randomised , double-blind , multicentre , 8-week parallel group study was conducted using DB05767 1200 mg/day compared with 4500 mg/day of slow release mesalazine ( mesalamine ) granules in patients with mild-to-moderately active ulcerative colitis . Disease activity was assessed at baseline and every 2 weeks for clinical response , and at baseline and 8 weeks by colonoscopy . RESULTS : One hundred and twenty patients at five centres in China were randomised and dosed . Clinical remission and response were seen in 21 % and 76 % of DB05767 -treated patients , and 16 % and 82 % of mesalazine-treated patients . By colonoscopy , remission and response were seen in 28 % and 74 % of DB05767 -treated patients and 24 % and 71 % of mesalazine-treated patients , respectively . There was no significant difference between the two treatment groups . CONCLUSION : DB05767 may be an efficacious alternative to mesalazine in ulcerative colitis . DB00188 -resistant myeloma cell lines : a role for mutated P28074 in preventing the accumulation of unfolded proteins and fatal ER stress . DB00188 is an effective agent for treating multiple myeloma ( MM ) . To investigate the underlying mechanisms associated with acquired resistance to this agent , we established two bortezomib-resistant MM cell lines , KMS-11/BTZ and OPM-2/BTZ , the 50 % inhibitory concentration values of which were respectively 24.7- and 16.6-fold higher than their parental cell lines . No activation of caspase and BH3-only proteins such as Noxa was noted in bortezomib-resistant cells after exposure to the drug . The accumulation of polyubiquitinated proteins was reduced in bortezomib-resistant cells compared with the parental cells , associated with avoidance of catastrophic ER stress as assessed by downregulation of P35638 expression . These resistant MM cells have a unique point mutation , G322A , in the gene encoding the proteasome beta5 subunit ( P28074 ) , likely resulting in conformational changes to the bortezomib-binding pocket of this subunit . KMS-11 parental cells transfected to express mutated P28074 also showed reduced bortezomib-induced apoptosis compared with those expressing wild-type P28074 or the parental cells . Expression of mutated P28074 was associated with the prevention of the accumulation of unfolded proteins . Thus , a fraction of MM cells may acquire bortezomib resistance by suppressing apoptotic signals through the inhibition of unfolded protein accumulation and subsequent excessive ER stress by a mutation of the P28074 gene . Innovative approaches for the development of antidepressant drugs : current and future strategies . Depression is a highly debilitating disorder that has been estimated to affect up to 21 % of the world population . Despite the advances in the treatment of depression with selective serotonin reuptake inhibitors ( SSRIs ) and serotonin and norepinephrine reuptake inhibitors ( SNRIs ) , there continue to be many unmet clinical needs with respect to both efficacy and side effects . These needs range from efficacy in treatment resistant patients , to improved onset , to reductions in side effects such as emesis or sexual dysfunction . To address these needs , there are numerous combination therapies and novel targets that have been identified that may demonstrate improvements in one or more areas . There is tremendous diversity in the types of targets and approaches being taken . At one end of a spectrum is combination therapies that maintain the benefits associated with SSRIs but attempt to either improve efficacy or reduce side effects by adding additional mechanisms ( P08908 , P28222 , P28221 , P28335 , alpha-2A ) . At the other end of the spectrum are more novel targets , such as neurotrophins ( P23560 , IGF ) , based on recent findings that antidepressants induce neurogenesis . In between , there are many approaches that range from directly targeting serotonin receptors ( P28335 , P50406 ) to targeting the multiplicity of potential mechanisms associated with excitatory ( glutamate , DB01221 , Q14416 , P41594 ) or inhibitory amino acid systems ( GABA ) or peptidergic systems ( neurokinin 1 , DB05394 1 , melanin-concentrating hormone 1 , V1b ) . The present review addresses the most exciting approaches and reviews the localization , neurochemical and behavioral data that provide the supporting rationale for each of these targets or target combinations . Modulation of the P22301 /IL-12 cytokine circuit by interferon-beta inhibits the development of epitope spreading and disease progression in murine autoimmune encephalomyelitis . IFN-beta has been shown to be effective in the treatment of multiple sclerosis ( MS ) . However , the primary mechanism by which IFN-beta mediates its therapeutic effect remains unclear . Recent studies indicate that under defined conditions , IFN-beta may downregulate DC expression of IL-12 . We and others have shown that IFN-beta may also downregulate P22301 . In light of the recently proposed paradigm that an P22301 /IL-12 immunoregulatory circuit controls susceptibility to autoimmune disease , we examined the effect of IFN-beta on the development and behavior of the autoreactive T cell repertoire during experimental autoimmune encephalomyelitis ( EAE ) , an animal model sharing many features with MS . SWXJ mice were immunized with the immunodominant p139-151 determinant of myelin proteolipid protein ( PLP ) , and at onset of EAE were treated every other day with IFN-beta . After eight weeks of treatment , we assessed autoreactivity and observed no significant IFN-beta effect on splenocyte proliferation or splenocyte production of P01579 , P60568 , P05112 , or P05113 in response to the priming determinant used to initiate disease . However , in IFN-beta treated mice , the cytokine profile in response to the priming immunogen was significantly skewed toward an increased production of P22301 and a concurrent decreased production of IL-12 . Moreover , the in vivo modulation of the P22301 /IL-12 immunoregulatory circuit in response to the priming immunogen was accompanied by an aborted development of epitope spreading . Our results indicate that IFN-beta induces a reciprocal modulation of the P22301 /IL-12 cytokine circuit in vivo . This skewed autoreactivity establishes an inflammatory microenvironment that effectively prevents endogenous self-priming thereby inhibiting the progression of disease associated with epitope spreading . Tumor-derived exosomes promote tumor progression and T-cell dysfunction through the regulation of enriched exosomal microRNAs in human nasopharyngeal carcinoma . Tumor-derived exosomes contain biologically active proteins and messenger and microRNAs ( miRNAs ) . These particles serve as vehicles of intercellular communication and are emerging mediators of tumorigenesis and immune escape . Here , we isolated 30-100 nm exosomes from the serum of patients with nasopharyngeal carcinoma ( NPC ) or the supernatant of TW03 cells . Increased circulating exosome concentrations were correlated with advanced lymphoid node stage and poor prognosis in NPC patients ( P < 0.05 ) . TW03-derived exosomes impaired T-cell function by inhibiting T-cell proliferation and Th1 and Th17 differentiation and promoting Treg induction by NPC cells in vitro . These results are associated with decreases in P29323 , P42224 , and P40763 phosphorylation and increases in P42229 phosphorylation in exosome-stimulated T-cells . TW03-derived exosomes increased the proinflammatory cytokines IL-1β , P05231 , and P22301 but decreased IFNγ , P60568 , and Q16552 release from P01730 + or CD8+ T-cells . Furthermore , five commonly over-expressed miRNAs were identified in the exosomes from patient sera or NPC cells : hsa-miR-24-3p , hsa-miR-891a , hsa-miR-106a-5p , hsa-miR-20a-5p , and hsa-miR-1908 . These over-expressed miRNA clusters down-regulated the Q9P0L2 signaling pathway to alter cell proliferation and differentiation . Overall , these observations reveal the clinical relevance and prognostic value of tumor-derived exosomes and identify a unique intercellular mechanism mediated by tumor-derived exosomes to modulate T-cell function in NPC .

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MH_train_1340

interacts_with DB00762?

multiple_choice

Novel agents that potentially inhibit irinotecan-induced diarrhea . DB00762 ( CPT-11 , 7-ethyl-10-[4-(1-piperidino)-1-piperidino] carbonyloxycamptothecin ) has exhibited clinical activities against a broad spectrum of carcinomas by inhibiting P11387 ( Topo I ) . However , severe and unpredictable dosing-limiting toxicities ( mainly myelosuppression and severe diarrhea ) hinder its clinical use . The latter consists of early and late-onset diarrhea , occurring within 24 hr or > or = 24 hr after CPT-11 administration , respectively . This review highlights novel agents potentially inhibiting CPT-11-induced diarrhea , which are designed and tested under guidance of disposition pathways and potential toxicity mechanisms . Early-onset diarrhea is observed immediately after CPT-11 infusion and probably due to the inhibition of acetylcholinesterase activity , which can be eliminated by administration of atropine . Late-onset diarrhea appears to be associated with intestinal exposure to SN-38 ( 7-ethyl-10-hydroxycamptothecin ) , the major active metabolite of CPT-11 , which may bind to Topo I and induce apoptosis of intestinal epithelia , leading to the disturbance in the absorptive and secretory functions of mucosa . CPT-11 and SN-38 may also stimulate the production of pro-inflammatory cytokines and prostaglandins ( PGs ) , thus inducing the secretion of Na(+) and Cl(-) . Early treatment of severe late-onset diarrhea with oral high-dose loperamide has decreased patient morbidity . Extensive studies have been conducted to identify other potential agents to ameliorate diarrhea in preclinical and clinical models . These include intestinal alkalizing agents , oral antibiotics , enzyme inducers , P-glycoprotein ( PgP ) inhibitors , cyclooxygenase-2 ( P35354 ) inhibitors , tumor necrosis factor-alpha ( P01375 ) inhibitors , or blockers of biliary excretion of SN-38 . Further studies are needed to identify the molecular targets associated with CPT-11 toxicity and safe and effective agents for alleviating CPT-11-induced diarrhea . Role of chronic inhibition of dopamine-metabolizing enzymes in the regulation of renal sodium and phosphate excretion in the rat remnant kidney . BACKGROUND/AIMS : The present study examined the effects of chronic selective or combined inhibition of type A monoamine oxidase ( MAO ) and catechol-O-methyltransferase ( P21964 ) on daily urinary excretion of dopamine and metabolites and on natriuresis and phosphaturia in 3/4 nephrectomized ( 3/4nx ) and Sham rats . METHODS : The 3/4nx and Sham rats were placed in metabolic cages and received the P21397 -selective inhibitor Ro-411049 ( 7.5 mg x kg(-1) bid ) and/or the P21964 -selective inhibitor DB03336 3-202 ( 30 mg x kg(-1) bid ) orally for 3 days during high sodium diet . RESULTS : Selective P21964 inhibition increased the urinary excretion of the deaminated metabolite ( 3,4-dihydroxyphenylacetic acid , DOPAC ) and decreased the urinary excretion of the methylated ( 3-methoxytyramine , 3-MT ) and deaminated plus methylated metabolite ( homovanillic acid , HVA ) in both groups . Selective P21397 inhibition increased the urinary excretion of 3-MT and reduced the urinary excretion of both DOPAC and HVA in either 3/4nx or Sham rats . Combined inhibition of P21397 and P21964 did not significantly change the urinary excretion of DOPAC and markedly decreased the urinary excretion of 3-MT and HVA in both groups . Selective or combined inhibition of P21397 and P21964 did not alter the daily urinary excretion of dopamine , sodium or phosphate in either 3/4nx or Sham rats . CONCLUSIONS : Chronic selective or combined inhibition of P21397 and P21964 is not of major importance in regulating the dopamine-dependent natriuresis and phosphaturia in either 3/4nx or Sham rats . DB00158 transport in mouse cumulus-oocyte complexes and preimplantation embryos . Endogenous folate stores are required in preimplantation embryos of several species , but how folates are accumulated and whether they can be replenished has not been determined . Folates are generally taken up into cells by specific transporters , mainly the reduced folate carrier RFC1 ( P41440 protein ) and the high-affinity folate receptors P15328 and P14207 . Quantitative RT-PCR showed that Slc19a1 mRNA was expressed in mouse cumulus-oocyte complexes ( COCs ) and oocytes , whereas Folr1 showed expression only in preimplantation embryos , increasing from the 2-cell stage onward . The mRNAs encoding Folr2 and the intestinal folate transporter Slc46a1 were not detected . DB00563 ( MTX ) , an antifolate often used as a model substrate for folate transport , exhibited saturable transport in COCs and in preimplantation embryos starting at the 2-cell stage . However , folate transport characteristics differed between COCs and embryos . In COCs , transport of MTX and the reduced folate leucovorin was inhibited by the anion transport inhibitor SITS that blocks RFC1 but was insensitive to dynasore , a specific dynamin inhibitor that instead inhibits folate receptor-receptor mediated endocytosis , whereas the opposite was found in 2-cell embryos and blastocysts . The inhibitor profile and transport properties of MTX and leucovorin in COCs correspond to established transport characteristics of RFC1 ( P41440 ) , whereas those in 2-cell embryos and blastocysts correspond with those of P15328 , consistent with the mRNA expression patterns . Considerable folate was accumulated in COCs via RFC1 , but the presence of cumulus cells did not enhance folate accumulation in the enclosed oocyte , indicating a lack of transfer from cumulus to oocyte . The promotion of iron-induced generation of reactive oxygen species in nerve tissue by aluminum . DB01370 is suspected to play a role in several neurological disorders . Reactive oxygen species ( ROS ) lead to oxidative stress , which is thought to be a possible mechanism for neurological damage . Interactions between aluminum and iron , a known promoter of prooxidant events , were studied in cerebral tissues using a fluorescent probe to measure rates of generation of ROS . Al2(SO4)3 alone failed to stimulate ROS production over a wide range of concentrations ( 50-1000 microM ) . The aluminum-deferrioxamine chelate in the absence of iron could also not potentiate ROS formation . However , Al2(SO4)3 potentiated FeSO4-induced ROS , with a maximal effect at 10 microM Fe and 500 microM Al . DB01575 , a hydrated aluminum silicate , did not potentiate iron-induced ROS formation . Ferritin had a minor stimulatory effect on ROS generation , but this was not potentiated by the concurrent presence of Al2(SO4)3 . P02787 had no effect on basal rates of ROS generation , but when Al2(SO4)3 was also present , ROS production was enhanced . It is concluded that : 1 . There is a potentiation of iron-induced ROS by aluminum salts ; 2 . Free or complexed aluminum alone is not a key producer of ROS ; and 3 . High rates of ROS production are unlikely to be owing to the displacement by aluminum iron from its biologically sequestered locations . Protective effects of heparin on endothelial cells in sepsis . This study aims to observe the protective effects of heparin on endothelial cells in sepsis and explore the involved signal pathway regulated by heparin . Methods Human vascular endothelial cells were treated by TNFα in vitro to simulate the inflammatory environment when sepsis occurred . They were intervened by heparin and the expression levels of soluble thrombomodulin ( sTM ) and serum activated protein C ( P25054 ) were detected by ELISA , the regulatory mechanism of heparin improving vascular endothelial cells injury induced by TNFα was detected by Western Blotting method , the methylation of histone in the gene promoter region of endothelial nitric oxide synthase ( P29474 ) and monocyte chemotactic protein-1 ( P13500 ) were detected using chromatin immunoprecipitation method . Results DB01109 could inhibit the secretion of sTM and P25054 protein and the expression of P13500 gene which involved in NF-κB signal pathway . Conclusions DB01109 could protect vascular endothelial cells from injury induced by TNFα and sepsis , the mechanisms were related with the effects of heparin on the histone methylation of promoter region and the regulation of heparin on the MAPK and NF-κB signal pathways . These results provide a theoretical basis for the application of heparin in the prevention and treatment of vascular disease related with sepsis . Q9GZP0 inhibition by DB05139 ameliorates tubulointerstitial fibrosis following experimental glomerulonephritis . BACKGROUND : Arresting or regressing kidney scarring is of major clinical relevance . Q9GZP0 ( Q9GZP0 ) is widely expressed in fibrotic kidneys . Administration of the Q9GZP0 neutralizing fully human monoclonal antibody DB05139 in the acute phase of progressive anti-Thy 1.1 glomerulonephritis reduced glomerular and secondary tubulointerstitial damage . METHODS : Using this model , we now assessed the effects of DB05139 ( n=15 ) vs irrelevant control IgG ( n=17 ) administered on days 17 , 28 and 35 after disease induction , i.e. after acute glomerular damage had subsided . RESULTS : In vitro , DB05139 inhibited the Q9GZP0 - but not the PDGF-B-induced proliferation of rat renal fibroblasts . Following the first DB05139 injection on day 17 , exposure to therapeutic levels was maintained until day 49 . Proteinuria in the DB05139 -treated group was transiently reduced between days 49 and 77 ( -19 to -23 % in comparison with the controls ; P < 0.05 ) . On day 100 , DB05139 treatment reduced the number of rats that had doubled their serum creatinine ( DB05139 : 40 vs controls : 71 % ; P < 0.05 ) . Compared with controls , the DB05139 animals , on day 100 , significantly lowered glomerular expression of vimentin and collagens as well as tubulointerstitial damage scores , interstitial fibrosis , vimentin and cortical Q9GZP0 mRNA levels . CONCLUSIONS : Q9GZP0 antagonism , even after the phase of acute glomerular damage , exerts beneficial effects on the course of tubulointerstitial damage , i.e. the final common pathway of most renal diseases . Pharmacogenetics in chemotherapy of colorectal cancer . Although in recent years , chemotherapeutic options for colorectal carcinoma have expanded , overall response rates are still too low , with high rates of toxicity . Pharmacogenetics aim at predicting both treatment response and adverse effects in individual patients . This review describes the current knowledge of pharmacogenetic markers in the systemic treatment of colorectal cancer . P22309 *28 leads to reduced conjugation of SN-38 , the active metabolite of irinotecan , resulting in an increased rate of adverse effects , especially neutropenia . To a lesser extent , increased DB00544 toxicity is predicted by Q12882 *2A . A variable number of tandem repeats polymorphism in the thymidylate synthase enhancer region , in combination with a single nucleotide polymorphism C > G , may predict poorer response to DB00544 . Efficacy of oxaliplatin is influenced by polymorphisms in components of DNA repair systems , such as P07992 and P18887 . Polymorphic changes in the endothelial growth factor receptor probably predict cetuximab efficacy . Furthermore , the antibody-depended cell-mediated cytotoxic effect of cetuximab may be reduced by polymorphisms in the immunoglobin G fragment C receptors . DB00112 efficacy is suspected to be influenced by polymorphisms in the P15692 gene and the hypoxia inducible factor 1alpha gene . Although the interpretation of pharmacogenetic studies is complicated , results imply a promising way of pretreatment prediction of chemotherapy efficacy and toxicity . P15121 inhibitor fidarestat attenuates leukocyte-endothelial interactions in experimental diabetic rat retina in vivo . PURPOSE : Dysregulation of the polyol pathway has been implicated as a major cause of diabetic retinopathy . The aldose reductase inhibitor fidarestat was recently reported to prevent retinal oxidative stress and overexpression of vascular endothelial growth factor ( P15692 ) protein in diabetic rats . In this study , we investigated the effect of fidarestat on leukocyte-endothelial cell interactions in an in vivo experimental model for diabetic retina . MATERIALS AND METHODS : Diabetes was induced in six-week-old male Long-Evans rats by intraperitoneal injection of streptozotocin ( Q11206 ) ( 75 mg/kg ) . The rats were divided into four experimental groups : non-diabetic control rats , untreated diabetic rats , and diabetic rats treated with a low ( 4 mg/kg/day ) or high ( 16 mg/kg/day ) oral dose of fidarestat . After four weeks of treatment , accumulated leukocytes in the retina were counted in vivo by acridine orange digital fluorography . Intercellular adhesion molecule-1 ( P05362 ) and P15692 -164 mRNA levels in the retina were analyzed using the quantitative reverse transcription-polymerase chain reaction . P05362 protein expression in the retina was investigated by immunohistochemistry . RESULTS : DB02021 treatment significantly decreased concentrations of sorbitol and fructose in the retinas of Q11206 -induced diabetic rats . Leukocyte accumulation in the retinas of fidarestat-treated rats was significantly less than in the untreated diabetic group ( P < 0.01 ) . DB02021 treatment significantly reduced the expression P05362 mRNA , but not P15692 -164 mRNA , in the retina of diabetic rats . Immunohistochemical study also revealed the suppressive effect of fidarestat on expression of P05362 . CONCLUSIONS : Oral administration of fidarestat attenuated leukocyte accumulation in the retina of Q11206 induced-diabetic rats , suggesting that fidarestat may have a therapeutic role in preventing the progression of diabetic retinopathy . Neurokinin-1 receptor blockade and murine lung tumorigenesis . RATIONALE : Analogous to the adenoma-carcinoma sequence in the colon , it has been proposed that adenocarcinoma ( AC ) in the lung arises from adenomatous hyperplasia that progresses through atypical adenomatous hyperplasia to AC . However , the data supporting this sequence are largely circumstantial and the almost impossible task of identifying these lesions before resection rules out any longitudinal study in humans . OBJECTIVES , METHODS , AND RESULTS : We show in mice that the loss of function of the neurokinin-1 receptor ( P25103 ) -due to either a pharmacologic or genetic manipulation-results in a sequence of morphologic changes in response to bleomycin treatment that precede the development of AC . We also demonstrate that a series of alterations in gene expression of proliferation markers ( i.e. , P12004 and Ki-67 ) and cell cycle regulators ( i.e. , P49789 , p53 , and P38936 ) characterizes the sequence of the precursor lesions . The loss of function of the P25103 results in changes of the apoptotic rate and in a delay of DNA break recovery of alveolar epithelial cells following bleomycin treatment . The P25103 blockade interferes with a caspase-independent pathway of apoptosis by affecting both the translocation of P22736 into the cytoplasm and the expression of some important Bcl2 family members such as Bcl2 and Bak . CONCLUSIONS : To our knowledge , this is the first model to demonstrate a role for P25103 in lung epithelial cell death and tumorigenesis . This animal model may provide new information on the biology of AC and will facilitate designing and testing of new therapeutic interventions . DB08816 increases adenosine plasma concentration in patients with an acute coronary syndrome . OBJECTIVES : This study aimed to investigate the impact of ticagrelor on adenosine plasma concentration ( P25054 ) in acute coronary syndrome ( ACS ) patients . BACKGROUND : DB08816 is a direct-acting Q9H244 -adenosine diphosphate receptor blocker . The clinical benefit of ticagrelor compared with clopidogrel in ACS patients suggests that the drug has non-platelet-directed properties . Animal and in vitro models suggested that the " pleiotropic " properties of ticagrelor may be related to an interaction with adenosine metabolism . METHODS : We prospectively randomized 60 ACS patients to receive ticagrelor or clopidogrel . The P25054 was measured by liquid chromatography . To assess the mechanism of P25054 variation , we measured adenosine deaminase concentration , adenosine uptake by red blood cells , and cyclic adenosine monophosphate production by cells overexpressing adenosine receptors . The Q9H244 -adenosine diphosphate receptor blockade was assessed by the vasodilator-stimulated phosphoprotein index . RESULTS : Patients receiving ticagrelor had significantly higher P25054 than patients receiving clopidogrel ( 1.5 μM [ interquartile range : 0.98 to 1.7 μM ] vs. 0.68 μM [ interquartile range : 0.49 to 0.78 μM ] ; p < 0.01 ) . The P25054 was not correlated with vasodilator-stimulated phosphoprotein ( p = 0.16 ) . Serum-containing ticagrelor inhibited adenosine uptake by red blood cells compared with clopidogrel or controls ( p < 0.01 for both comparisons ) . DB00640 deaminase activity was similar in serum of patients receiving clopidogrel or ticagrelor ( p = 0.1 ) . DB08816 and clopidogrel had no direct impact on adenosine receptors ( p = not significant ) . CONCLUSIONS : DB08816 increases P25054 in ACS patients compared with clopidogrel by inhibiting adenosine uptake by red blood cells . Inflammatory and motor responses by tachykinins in the guinea-pig oesophageal sphincter . 1. The aim of this study was to characterize the tachykinin receptors involved in producing plasma protein extravasation and contractile responses of the guinea-pig oesophageal sphincter . 2 . In anaesthetized guinea-pigs , the selective tachykinin P25103 agonist [Sar9, DB00134 (O2)11] DB05875 produced plasma protein extravasation ( PPE ) which was not affected by the treatment with the tachykinin P21452 antagonist MEN 10627 ( 1 micromol kg(-1) i.v. ) or the histamine H1-receptor antagonist , diphenhydramine ( 34.5 micromol kg(-1) i.v. ) . However , the [Sar9, DB00134 (O2)11] DB05875 -induced PPE was blocked by the previous administration of the peptide tachykinin P25103 antagonist FK 888 or by the non-peptide antagonist DB05790 , yielding ED50 values of 1.1 +/- 0.2 and 0.01 +/- 0.004 micromol kg(-1) i.v. , respectively . 3 . The tachykinin NK-2 or P29371 agonists [beta-Ala8]neurokinin A ( 4-10 ) or [MePhe7]neurokinin B , respectively , produced a weak PPE at high doses . The effect of [MePhe7]neurokinin B-induced PPE was inhibited by DB05790 . 4 . In the guinea-pig isolated oesophageal sphincter , [Sar9, DB00134 (O2)11] DB05875 did not exert any contractile effect up to 10 microM . The selective tachykinin P21452 agonist ( [beta-Ala8]neurokinin A ( 4-10 ) , produced concentration-dependent contractions ( pD2 = 7.6 +/- 0.03 ) which were inhibited by the selective tachykinin P21452 antagonist , MEN 10627 ( pA2 = 8.6 +/- 0.1 ) . Also , the tachykinin P29371 selective agonist [MePhe7]neurokinin B induced concentration-dependent contractile responses , but these responses were inhibited by MEN 10627 . 5 . Altogether , these data indicate that the stimulation of tachykinin P25103 produces a vascular inflammatory response , while activation of tachykinin NK-2 receptors mediate the contraction of the guinea-pig oesophageal sphincter . Q9H244 receptors play a significant role in the development of platelet microaggregation in patients with diabetes . Ninety-eight diabetic patients ( type 2 ) were studied together with 24 healthy normotensive controls . Microaggregates ( particle scale , < 25 microm ) of platelets were detected by a laser scattering system . Microaggregates in the control group showed a time-dependent reversible change ; however , they existed continuously in 82 of 98 diabetic patients . When platelets of diabetics were stimulated by a shear stress alone without ADP , 74 also showed spontaneous and irreversible microaggregates even though they were not observed in all control subjects . In control subjects , microaggregates were inhibited by MRS2279 ( a P47900 antagonist ) , but not AR-C69931MX ( a Q9H244 antagonist ) . However , AR-C69931MX prevented irreversible microaggregates in diabetic patients . When either aspirin or ticlopidine was administered to diabetic patients with irreversible microaggregates , both drugs significantly decreased microaggregates induced by a low dose of ADP . DB00208 additionally reduced the microaggregates induced by shear stress alone . In conclusion , microaggregates of platelets via Q9H244 receptors could play a key role in the hypersensitivity of platelets in diabetic patients , and the measurement of microaggregation could be a useful marker to estimate of thrombogenesis . These findings present a possible new means for patients with diabetes to prevent ischemic events . P02787 subtypes and variants in Germany ; further evidence for a Tf null allele . Isoelectric focusing ( IEF ) with carrier ampholytes was used for the determination of transferrin C subtypes and transferrin B and D variants in a sample of 1125 unrelated individuals from Southern Germany . The observed TfC allele frequencies were Tf*C1 = 0.7872 , Tf* P06681 = 0.1365 , and Tf* P01024 = 0.0675 . The rare C subtype P13671 was observed twice . A new C subtype , called Q99622 , was observed and identified by IEF with immobilized pH gradients . The rare C subtypes C4 and Q99618 were also studied by this method . TfB and TfD variants were found with a heterozygous frequency of 1.53 % . One new TfD was found which is located between D1 and D2 and therefore named D1-2 . Evidence for a Tf null allele was obtained in a child and the putative father ; they were considered to be heterozygous for an allele Tf0 . The theoretical exclusion rate for paternity examinations was calculated for the Tf system and found to be 17.95 % . Enhanced sensitivity to irinotecan by Cdk1 inhibition in the p53-deficient HT29 human colon cancer cell line . Mutations in the tumor-suppressor gene p53 have been associated with advanced colorectal cancer ( CRC ) . DB00762 ( CPT-11 ) , a P11387 inhibitor , has been recently incorporated to the adjuvant therapy . Since the DNA-damage checkpoint depends on p53 activation , the status of p53 might critically influence the response to CPT-11 . We analysed the sensitivity to CPT-11 in the human colon cancer cell line HT29 ( mut p53 ) and its wild-type (wt)-p53 stably transfected subclone HT29-A4 . Cell-cycle analysis in synchronised cells demonstrated the activation of transfected wt-p53 and a P38936 ( P38936 /CIP1)-dependent cell-cycle blockage in the S phase . Activated wt-p53 increased apoptosis and enhanced sensitivity to CPT-11 . In p53-deficient cells , cDNA-macroarray analysis and western blotting showed an accumulation of the cyclin-dependent kinase (cdk)1/cyclin B complex . Subsequent p53-independent activation of the cdk-inhibitor ( cdk-I ) P38936 ( P38936 /CIP1) prevented cell-cycle progression . Cdk1 induction was exploited in vivo to improve the sensitivity to CPT-11 by additional treatment with the cdk-I P99999 -202 . We demonstrate a gain of sensitivity to CPT-11 in a p53-mutated colon cancer model either by restoring wild-type p53 function or by sequential treatment with cdk-Is . Considering that mutations in p53 are among the most common genetic alterations in CRC , a therapeutic approach specifically targeting p53-deficient tumors could greatly improve the treatment outcomes . Evaluation of the toxicity of the dopaminergic neurotoxins MPTP and P25189 + in PC12 pheochromocytoma cells : binding and biological studies . This study was designed to investigate the toxicity of both MPTP and P25189 + using some simple cell systems , such as PC12 and P13671 cultures , as models . Exposure of PC12 cells to 0.5 mM MPTP for 72 h resulted in a 50 % cell loss with respect to the control cells , and clorgyline , a P21397 inhibitor , antagonized this toxic effect . Higher concentrations of MPTP demonstrated only a weak cytostatic effect on P13671 cells . Moreover , P25189 + showed a toxic effect which was 100 times more evident than MPTP toxicity in the PC12 . We found a single , saturable class of [3H] P25189 + binding sites with a relatively high affinity both in PC12 and P13671 cell lines . Moreover , the most susceptible cell line towards the toxic effects of both MPTP and P25189 + , i.e. PC12 , has the higher number of P25189 + binding sites . Our results suggest that MPTP can be toxic not only via P27338 , but also via P21397 activity and we propose PC12 as a model to study the intracellular mechanisms of MPTP and P25189 + toxicity . Different histological types of non-small cell lung cancer have distinct folate and DNA methylation levels . Aberrant DNA methylation is a commonly observed epigenetic change in lung cancer . DB00158 has been suggested to play a role in the homeostasis of DNA methylation and has also been implicated in cancer chemotherapy . We investigated a possible role for folate in DNA methylation by measuring folate concentrations in tumors and adjacent normal tissues from 72 non-small cell lung cancer ( NSCLC ) patients . These were compared to DNA methylation levels and to clinicopathological features . DB00158 concentrations were determined as the sum of 5,10-methylenetetrahydrofolate and DB00116 . The MethyLight assay was used to quantitate methylation in promoter regions of P16(CDKN2A) , P25054 , P55290 , P10826 , Q9NS23 , Q13761 , and P15172 . Methylation of LINE-1 repeats was used as a surrogate for global methylation . DB00158 levels in tumors correlated positively with LINE-1 , P55290 , and Q13761 methylation . DB00158 concentrations and methylation of LINE-1 , Q9NS23 , and Q13761 were significantly higher in adenocarcinoma compared to squamous cell carcinoma ( SCC ) . Two sets of array-based data retrieved from the Gene Expression Omnibus consistently showed that expression of P15328 , a folate transport enzyme , and Q92820 , an enzyme that prevents folate retention , were higher and lower , respectively , in adenocarcinomas compared to SCC . This was independently validated by quantitative RT-PCR in 26 adenocarcinomas and 13 SCC . Our results suggest that folate metabolism plays a role in aberrant DNA methylation in NSCLC . The histological subtype differences in folate concentration and DNA methylation observed here were associated with distinct expression patterns for folate metabolizing enzymes . These findings may have clinical applications for histology-directed chemotherapy with fluoropyrimidine and anti-folates in NSCLC . Vasopeptidase inhibition exhibits endothelial protection in salt-induced hypertension . DB00886 represents a new class of drugs capable of inhibiting both P12821 and neutral endopeptidase 24.11 , the so-called vasopeptidase inhibitors . It therefore contributes to neurohumoral modulation , which might improve endothelial function in cardiovascular diseases . This study investigated the effect of omapatrilat in comparison to the P12821 inhibitor captopril on systolic blood pressure and endothelial function in salt-induced hypertension . Dahl salt-sensitive rats ( n=6/group ) on standard or salt-enriched ( 4 % NaCl ) chow were treated for 8 weeks with either omapatrilat ( 36+/-4 mg/kg per day ) , captopril ( 94+/-2 mg/kg per day ) , or placebo . Aortic rings were then isolated and suspended in organ chambers for isometric tension recording . Systolic blood pressure of salt-fed , placebo-treated animals increased to 196+/-6 mm Hg , which was prevented by omapatrilat ( 162+/-5 mm Hg , P < 0.05 ) and captopril ( 164+/-7 mm Hg , P < 0.05 ) to a comparable degree . In control rats , acetylcholine ( 10(-10) to 10(-5) mol/L ) induced endothelium-dependent relaxation ( 97+/-4 % ) , which was reduced by high-salt diet to 30+/-5 % ( P < 0.005 ; n=6 ) . DB00886 improved relaxation to a greater extent ( 86+/-5 % ) than did captopril ( 57+/-6 % ; P < 0.05 ) . P29474 protein expression and aortic nitrite/nitrate content were reduced in hypertensive rats and improved by both omapatrilat and captopril . Aortic endothelin-1 levels were increased in salt-fed animals and unaffected by omapatrilat or captopril . These data suggest that despite comparable blood pressure , omapatrilat is superior to captopril in improving endothelium-dependent relaxation in salt-sensitive hypertension . Lessons learned from the irinotecan metabolic pathway . DB00762 , a camptothecin analogue , is a prodrug which requires bioactivation to form the active metabolite SN-38 . SN-38 acts as a P11387 poison . DB00762 has been widely used in the treatment of metastatic colorectal cancer , small cell lung cancer and several other solid tumors . However , large inter-patient variability in irinotecan and SN-38 disposition , as well as severe but unpredictable diarrhea limits the clinical potential of irinotecan . Intense clinical pharmacology studies have been conducted to elucidate its complicated metabolic pathways and to provide scientific rationale in defining strategies to optimize drug therapy . DB00762 is subjected to be shunted between P08684 mediated oxidative metabolism to form two inactive metabolites P25054 or NPC and tissue carboxylesterase mediated hydrolysis to form SN-38 which is eventually detoxified via glucuronidation by P22309 to form SN-38G . The pharmacology of this compound is further complicated by the existence of genetic inter-individual differences in activation and deactivation enzymes of irinotecan ( e.g. , P08684 , P20815 , P22309 ) and sharing competitive elimination pathways with many concomitant medications , such as anticonvulsants , St . John 's Wort , and ketoconazole . Efflux of the parent compound and metabolites out of cells by several drug transporters ( e.g. , Pgp , Q9UNQ0 , MRP1 , Q92887 ) also occurs . This review highlights the latest findings in drug activation , transport mechanisms , glucuronidation , and CYP3A-mediated drug-drug interactions of irinotecan in order to unlock some of its complicated pharmacology and to provide ideas for relevant future studies into optimization of this promising agent . [ Quantitative analysis of P11387 activity in human and rat glioma : characterization and mechanism of resistance to antitopoisomerase chemical , camptothecin-11 ] . DB00762 ( CPT-11 ) is a new derivation of camptothecin , a plant alkaloid antitumor agent . Previous studies indicated that antitumor activity of CPT-11 was mediated through interaction of the drugs with its target enzyme , P11387 ( topo I ) . In this study , we studied the relation between sensitivity to CPT-11 and topo I activity of glioma cells . Furthermore , we established CPT-11 resistant cell lines in order to elucidate potential mechanisms of drug resistance . A clear correlation between the sensitivities to CPT-11 and topo I activities in surgical glioma specimens was demonstrated . Activities of topo I in CPT-11 sensitive group ( IC50 values for CPT-11 ; < 50 micrograms/ml ) tended to be higher than those in CPT-11 resistant group ( IC50 values ; > or = 50 ) . Topo I activity may serve as a novel marker to predict the sensitivity of gliomas to topo inhibitors . CPT-11 resistance cell lines ( T98G/CPT-11 and P13671 ) respectively exhibit a 5.4- and 7.3-fold increase in resistance to CPT-11 . No differences in topo I activity and intracellular accumulation of CPT-11 were observed between parent and CPT-11 resistant lines . On the other hand , topo I from T98G/CPT-11 and P13671 /CPT-11 cells were at least 4- and 2-fold resistant to the inhibitory effect of the CPT-11 on the relaxation activity of topo I in comparison with their parent lines . This enzymological difference may be responsible for the resistance to CPT-11 . Platelet-derived growth factor-D induces expression of cyclooxygenase-2 in rat mesangial cells through activation of PI3K/ P31749 and PKCs . Platelet-derived growth factor ( PDGF ) -D is suggested to be a key factor in the development of several renal pathologies , including mesangioproliferative glomerulonephritis . Cyclooxygenase ( P36551 ) -2 is a protein involved in the biosynthesis of inflammatory prostaglandins . In this study , we investigated the effect of Q9GZP0 on the regulation of P35354 expression in rat mesangial cells ( RMCs ) . Treatment with Q9GZP0 induced P35354 at both the protein and mRNA levels in RMCs , suggesting that the Q9GZP0 -mediated induction of P35354 is due to P35354 transcriptional upregulation . Q9GZP0 treatment also led to a rapid but transient activation of P31749 and extracellular signal regulated kinase ( P29323 ) -1/2 . Activities of JNK-1/2 and p38 MAPK , however , were not influenced by Q9GZP0 in RMCs . Markedly , pharmacological inhibition studies showed that pretreatment with LY294002 ( a PI3K/ P31749 inhibitor ) or GF109203X ( a pan-PKC inhibitor ) suppressed the Q9GZP0 -induced expression of P35354 protein and mRNA , while pretreatment with PD98059 ( an P27361 /2 inhibitor ) or P50391 ( an Src inhibitor ) had no effect on it . These findings collectively demonstrate for the first time that Q9GZP0 induces P35354 by transcriptional upregulation in RMCs and the induction is largely related to PI3K/ P31749 and PKCs activities . DB06243 ( DB06243 ) prevents progression of pancreatic cancer by modulating ornithine decarboxylase signaling . P11926 ( ODC ) is the key rate-limiting enzyme in the polyamine synthesis pathway and it is overexpressed in a variety of cancers . We found that polyamine synthesis and modulation of ODC signaling occurs at early stages of pancreatic precursor lesions and increases as the tumor progresses in Kras-activated p48(Cre/+)-LSL-Kras(G12D/+) mice . Interest in use of the ODC inhibitor eflornithine ( DB06243 ) as a cancer chemopreventive agent has increased in recent years since ODC was shown to be transactivated by the c-myc oncogene and to cooperate with the ras oncogene in malignant transformation of epithelial tissues . We tested the effects of DB06243 on pancreatic intraepithelial neoplasias ( PanIN ) and their progression to pancreatic ductal adenocarcinoma ( PDAC ) in genetically engineered Kras mice . The Kras(G12D/+) mice fed DB06243 at 0.1 % and 0.2 % in the diet showed a significant inhibition ( P < 0.0001 ) of PDAC incidence compared with mice fed control diet . Pancreatic tumor weights were decreased by 31 % to 43 % ( P < 0.03-0.001 ) with both doses of DB06243 . DB06243 at 0.1 % and 0.2 % caused a significant suppression ( 27 % and 31 % ; P < 0.02-0.004 ) of PanIN 3 lesions ( carcinoma in situ ) . DB06243 -treated pancreas exhibited modulated ODC pathway components along with decreased proliferation and increased expression of P38936 /p27 as compared with pancreatic tissues derived from mice fed control diet . In summary , our preclinical data indicate that DB06243 has potential for chemoprevention of pancreatic cancer and should be evaluated in other PDAC models and in combination with other drugs in anticipation of future clinical trials .

MH_train_1341

MH_train_1341

MH_train_1341

interacts_with DB08827?

multiple_choice

Immunobiotic Lactobacillus strains augment Q96P20 expression in newborn and adult porcine gut-associated lymphoid tissues . We isolated cDNA encoding porcine nucleotide-binding domain-like receptor family , pryin domain containing 3 ( Q96P20 ) from Peyer 's patches . The complete nucleotide open reading frame of porcine Q96P20 contains 3108-bp encoding a deduced polypeptide of 1036-amino acid residues . The porcine Q96P20 amino acid sequence is more similar to the longest isoform of human than the mouse counterpart . The predicted amino acid sequence of porcine Q96P20 presented nine C-terminal leucine-rich repeat domains . In newborn swine , the expression of Q96P20 was detected at higher levels in spleen and mesenteric lymph nodes , while lower levels were observed in intestinal tissues . In adult swine , Q96P20 was strongly expressed in Peyer 's patches and the mesenteric lymph nodes , and the expression level in the lower intestinal tissues was comparable to that in spleen . Toll-like receptor and nucleotide-binding domain ligands , as well as Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus gasseri , enhanced Q96P20 expression in gut-associated lymphoid tissues ( P07902 ) of newborn and adult swine . Our results should aid in understanding the intestinal immunoregulatory mechanisms underlying Q96P20 activation and the priming ability of immunobiotic lactic acid bacteria in porcine P07902 . Beyond statins : new lipid lowering strategies to reduce cardiovascular risk . Statins are the first-line therapy in LDL- DB04540 ( LDL-C ) reduction and its clinical use has contributed to significant prevention and treatment of atherosclerotic vascular disease . Yet , a significant proportion of patients remain at high risk . Recently , a number of new therapies have been developed to further lower LDL-C . These agents may provide clinical benefit on top of statin therapy in patients with high residual risk , severe hypercholesterolemia or as an alternative for patients who are intolerant to statins . We review four novel approaches based on the inhibition of proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) , apolipoprotein-B100 ( apoB ) , Cholesteryl ester transport protein ( P11597 ) and microsomal triglyceride transfer protein ( P55157 ) . ApoB and P55157 inhibitors ( DB05528 and DB08827 ) are indicated only for homozygous familial hypercholesterolemia patients . The results of ongoing trials with P11597 and Q8NBP7 inhibitors may warrant a wider employment in different categories of patients at high risk for cardiovascular disease . DB00428 -induced increase in cholesterol ester transfer protein ( P11597 ) and its reversal by insulin in transgenic mice expressing human P11597 . High plasma triacylglycerol and low high-density lipoprotein levels are risk factors for cardiovascular disease in diabetes . Plasma high-density lipoprotein levels are regulated by cholesterol ester transfer protein ( P11597 ) . The regulation of P11597 under diabetic conditions is not clear , and this is due to a lack of appropriate models . We used transgenic mice expressing human P11597 to study the regulation of this protein under type-1 diabetic conditions and further investigated whether insulin reverses the effect of diabetes . Mice expressing human P11597 under the control of its natural flanking region and age-matched littermates not expressing this protein were made diabetic by injecting streptozotocin , and the reversal of diabetes was assessed by injecting insulin . The plasma total cholesterol , low-density lipoprotein-cholesterol , and triacylglycerol concentrations were elevated , whereas high-density lipoprotein-cholesterol concentrations were reduced after the onset of diabetes . P01308 injection partially recovered this effect . The plasma cholesterol ester transfer activity , P11597 mass , and hepatic P11597 mRNA abundance were significantly higher in diabetic mice that were partially restored by insulin administration . There was a strong correlation between high-density lipoprotein-cholesterol concentrations and cholesterol ester transfer activity . These results suggest that an increase in P11597 under diabetic conditions might be a major factor responsible for increased incidence of diabetes-induced atherosclerosis . A common binding site on the microsomal triglyceride transfer protein for apolipoprotein B and protein disulfide isomerase . The assembly of triglyceride-rich lipoproteins requires the formation in the endoplasmic reticulum of a complex between apolipoprotein B ( apoB ) , a microsomal triglyceride transfer protein ( P55157 ) , and protein disulfide isomerase ( P07237 ) . In the P55157 complex , the amino-terminal region of P55157 ( residues 22-303 ) interacts with the amino-terminal region of apoB ( residues 1-264 ) . Here , we report the identification and characterization of a site on apoB between residues 512 and 721 , which interacts with residues 517-603 of P55157 . P07237 binds in close proximity to this apoB binding site on P55157 . The proximity of these binding sites on P55157 for P07237 and amino acids 512-721 of apoB was evident from studies carried out in a yeast two-hybrid system and by co-immunoprecipitation . The expression of P07237 with P55157 and apoB16 ( residues 1-721 ) in the baculovirus expression system reduced the amount of P55157 co-immunoprecipitated with apoB by 73 % . The interaction of residues 512-721 of apoB with P55157 facilitates lipoprotein production . Mutations of apoB that markedly reduced this interaction also reduced the level of apoB-containing lipoprotein secretion . DB04998 inhibits activation of nuclear factor-kappaB ( NF-kappaB ) by forming a complex with NF-kappaB essential modulator ( Q9Y6K9 ) and nucleolin . DB04998 , also known as DB04998 , is an experimental anticancer drug that recently entered human clinical trials . It is a member of a novel class of antiproliferative agents known as G-rich oligonucleotides ( P09341 ) , which are non-antisense , guanosine-rich phosphodiester oligodeoxynucleotides that form stable G-quadruplex structures . The biological activity of GROs results from their binding to specific cellular proteins as aptamers . One important target protein of GROs has been previously identified as nucleolin , a multifunctional protein expressed at high levels by cancer cells . Here , we report that DB04998 also associates with nuclear factor-kappaB ( NF-kappaB ) essential modulator ( Q9Y6K9 ) , which is a regulatory subunit of the inhibitor of kappaB ( IkappaB ) kinase ( IKK ) complex , and also called IKKgamma . In the classic NF-kappaB pathway , the IKK complex is required for phosphorylation of P25963 and subsequent activation of the transcription factor NF-kappaB . We found that treatment of cancer cells with DB04998 inhibits IKK activity and reduces phosphorylation of P25963 in response to tumor necrosis factor-alpha stimulation . Using a reporter gene assay , we showed that DB04998 blocks both tumor necrosis factor-alpha-induced and constitutive NF-kappaB activity in human cancer cell lines derived from cervical , prostate , breast , and lung carcinomas . In addition , we showed that , in DB04998 -treated cancer cells , Q9Y6K9 is coprecipitated by nucleolin , indicating that both proteins are present in the same complex . Our studies suggest that abrogation of NF-kappaB activity may contribute to the anticancer effects of DB04998 and that nucleolin may play a previously unknown role in regulating the NF-kappaB pathway . DB08827 . Aegerion Pharmaceuticals is developing lomitapide , a small-molecule , microsomal triglyceride transfer protein ( P55157 ) inhibitor , for the treatment of both familial and primary hypercholesterolemia . Oral , once-daily lomitapide will be targeted at patients resistant to P04035 inhibitors ( statins ) either due to abnormalities in liver function or to discontinuation because of muscle pain . An oral formulation of lomitapide is in phase III development for homozygous familial hypercholesterolemia ( hyperlipoproteinemia type IIa ) in the US , Canada , Italy , and South Africa . This review discusses the key development milestones and therapeutic trials of this drug . Knockdown of Q03426 does not lead to changes in Q96P20 expression or activation . BACKGROUND : Mutations in the Mevalonate Kinase gene ( Q03426 ) are causes of a rare autoinflammatory disease : Mevalonate Kinase Deficiency and its more acute manifestation , Mevalonic Aciduria . The latter is characterized , among other features , by neuroinflammation , developmental delay and ataxia , due to failed cerebellar development or neuronal death through chronic inflammation . Pathogenesis of neuroinflammation in Mevalonate Kinase Deficiency and Mevalonic Aciduria has not yet been completely clarified , however different research groups have been suggesting the inflammasome complex as the key factor in the disease development . A strategy to mimic this disease is blocking the mevalonate pathway , using P04035 inhibitors ( Statins ) , while knock-out mice for Mevalonate Kinase are non-vital and their hemyzygous ( i.e only one copy of gene preserved ) littermate display almost no pathological features . FINDINGS : We sought to generate a murine cellular model closely resembling the pathogenic conditions found in vivo , by direct silencing of Mevalonate Kinase gene . Knockdown of Mevalonate Kinase in a murine microglial cellular model ( BV-2 cells ) results in neither augmented Q96P20 expression nor increase of apoptosis . On the contrary , statin treatment of BV-2 cells produces an increase both in Mevalonate Kinase and Q96P20 expression . CONCLUSIONS : MKD deficiency could be due or affected by protein accumulation leading to Q96P20 activation , opening novel questions about strategies to tackle this disease . Effects of MLN518 , a dual P36888 and P10721 inhibitor , on normal and malignant hematopoiesis . Internal tandem duplications ( ITDs ) of the P07333 -like tyrosine kinase 3 ( P36888 ) receptor tyrosine kinase are found in approximately 30 % of patients with acute myelogenous leukemia ( AML ) and are associated with a poor prognosis . P36888 ITD mutations result in constitutive kinase activation and are thought to be pathogenetically relevant , implicating P36888 as a plausible therapeutic target . MLN518 ( formerly CT53518 ) is a small molecule inhibitor of the P36888 , P10721 , and platelet-derived growth-factor receptor ( P09619 ) tyrosine kinases with significant activity in murine models of P36888 ITD-positive leukemia . Given the importance of P36888 and P10721 for normal hematopoietic progenitor cells , we analyzed the effect of MLN518 on murine hematopoiesis under steady-state conditions , after chemotherapy-induced myelosuppression , and during bone marrow transplantation . In these assays , we show that MLN518 has mild toxicity toward normal hematopoiesis at concentrations that are effective in treating P36888 ITD-positive leukemia in mice . We also demonstrate that MLN518 preferentially inhibits the growth of blast colonies from P36888 ITD-positive compared with ITD-negative patients with AML , at concentrations that do not significantly affect colony formation by normal human progenitor cells . In analogy to imatinib mesylate in P11274 - P00519 -positive acute leukemia , MLN518-induced remissions may not be durable . Our studies provide the basis for integrating this compound into chemotherapy and transplantation protocols . Gene polymorphisms in P02649 , NOS3 , and P11150 genes may be risk factors for cardiac adverse events after primary CABG . INTRODUCTION : Coronary artery disease progression after primary coronary artery bypass grafting may , beside classical atherosclerosis risk factors , be depending on genetic predisposition . METHODS : We investigated 192 CABG patients ( 18 % female , age : 60.9 +/- 7.4 years ) . Clinically cardiac adverse events were defined as need for reoperation ( n = 88 ; 46 % ) , reintervention ( n = 58 ; 30 % ) , or angina ( n = 89 ; 46 % ) . Mean follow-up time measured 10.1 +/- 5.1 years . Gene polymorphisms ( ApoE , NOS3 , P11150 , P11597 , SERPINE-1 , P00734 ) were investigated separately and combined ( gene risk profile ) . RESULTS : Among classical risk factors , arterial hypertension and hypercholesterinemia significantly influenced CAD progression . Single ApoE , NOS3 and P11150 polymorphisms provided limited information . Patients missing the most common ApoE epsilon 3 allele ( 5,2 % ) , showed recurrent symptoms ( p = 0,077 ) and had more frequently reintervention ( p = 0,001 ) . NOS3 a allele was associated with a significant increase for reintervention ( p = 0,041 ) and recurrent symptoms ( p = 0,042 ) . Homozygous P11150 patients had a higher reoperation rate ( p = 0.049 ) . A gene risk profile enabled us to discriminate between faster and slower occurrence of cardiac adverse events ( p = 0.0012 ) . CONCLUSION : Single P02649 , P11150 and NOS3 polymorphisms permitted limited prognosis of cardiac adverse events in patients after CABG . Risk profile , in contrast , allowed for risk stratification . DB08827 : A novel agent for the treatment of homozygous familial hypercholesterolemia . PURPOSE : The pharmacology , pharmacokinetics , and clinical efficacy and safety of lomitapide in the management of homozygous familial hypercholesterolemia ( HoFH ) are reviewed . SUMMARY : DB08827 ( Juxtapid , Aegerion Pharmaceuticals ) is an oral microsomal triglyceride transfer protein ( P55157 ) inhibitor indicated for the treatment of patients with HoFH , a rare form of hypercholesterolemia that can lead to premature atherosclerotic disease . In clinical trials , the use of lomitapide alone or in combination with other lipid-lowering modalities reduced plasma concentrations of low-density lipoprotein cholesterol ( LDL-C ) by a mean of more than 50 % . DB08827 is associated with significant gastrointestinal adverse effects and increases in hepatic fat levels . DB08827 undergoes hepatic metabolism via cytochrome P-450 ( CYP ) isoenzyme 3A4 and interacts with P08684 substrates including atorvastatin and simvastatin ; dose adjustment is recommended when lomitapide is used concurrently with these agents . In patients receiving concomitant warfarin , the International Normalized Ratio ( INR ) should be closely monitored , as lomitapide use may increase INR values . The recommended initial dosage of lomitapide is 5 mg once daily , with subsequent upward dose adjustment at specified intervals according to tolerability . DB08827 is contraindicated in patients with moderate-to-severe liver disease , patients with sustained abnormal liver function tests , patients taking strong or moderate P08684 inhibitors , and pregnant patients . CONCLUSION : DB08827 is an oral P55157 inhibitor approved for the treatment of HoFH . This agent appears to be a realistic option for patients with HoFH who are unable to attain their LDL-C goal or can not tolerate statin therapy . Protein disulphide isomerase is required for signal peptide peptidase-mediated protein degradation . The human cytomegalovirus glycoprotein US2 induces dislocation of MHC class I heavy chains from the endoplasmic reticulum ( ER ) into the cytosol and targets them for proteasomal degradation . Q8TCT9 ( Q8TCT9 ) has been shown to be integral for US2-induced dislocation of MHC class I heavy chains although its mechanism of action remains poorly understood . Here , we show that knockdown of protein disulphide isomerase ( P07237 ) by RNA-mediated interference inhibited the degradation of MHC class I molecules catalysed by US2 but not by its functional homolog US11 . Overexpression of the substrate-binding mutant of P07237 , but not the catalytically inactive mutant , dominant-negatively inhibited US2-mediated dislocation of MHC class I molecules by preventing their release from US2 . Furthermore , P07237 associated with Q8TCT9 independently of US2 and knockdown of P07237 inhibited Q8TCT9 -mediated degradation of CD3delta but not Q9BUN8 -dependent degradation of P13569 DeltaF508 . Together , our data suggest that P07237 is a component of the Q8TCT9 -mediated ER-associated degradation machinery . P01308 -like growth factor-I is a serum component stimulating growth of human neuroblastoma . Human non-autocrine neuroblastoma cells SK-N-SH and LF require serum for proliferation in vitro . We wished to determine the role of serum-borne insulin-like growth factor I ( P05019 ) as mitogen for these cells . Introduction of the monoclonal antibody alpha-IR3 against human P08069 reduced proliferation in the presence of fetal bovine serum ( FBS ) . P05019 ( 5 nM ) was as effective as FBS ( 10 % ) in stimulating proliferation . DB00071 mimicked the effects of P05019 , but at a 1000-fold higher concentration . The antibody alpha-IR3 reduced growth stimulated by P05019 more effectively than growth stimulated by insulin . Thus , proliferation of human non-autocrine neuroblastoma cells can be effectively manipulated by exogenous P05019 . Microsomal transfer protein ( P55157 ) inhibition-a novel approach to the treatment of homozygous hypercholesterolemia . Homozygous familial hypercholesterolemia ( HoFH ) represents the most severe lipoprotein disorder , generally attributable to mutation(s) of the low-density lipoprotein receptor ( LDL-R ) , i.e. autosomal dominant hypercholesterolemia type 1 ( P07327 ) . Much lower percentages are due to alterations of apolipoprotein B ( P00325 ) , or gain-of-function mutations of proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) ( P00326 ) . In certain geographical areas a significant number of patients may be affected by an autosomal recessive hypercholesterolemia ( Q5SW96 ) . Mutations may be also combined ( two mutations of the same gene , compound heterozygosity ) , or two in different genes ( double heterozygosity ) . Among the most innovative therapeutic approaches made available recently , inhibitors of the microsomal transfer protein ( P55157 ) system have shown a high clinical potential . P55157 plays a critical role in the assembly/secretion of very-low-density lipoproteins ( VLDL ) , and its absence leads to apo B deficiency . P55157 antagonists dramatically lower LDL-cholesterol ( LDL-C ) in animals , although a reported increase of liver fat delayed their clinical development . DB08827 , the best-studied P55157 inhibitor , reduces LDL-C by 50 % or more in HoFH patients , with modest , reversible , liver steatosis . Recent US approval has confirmed an acceptable tolerability , provided patients adhere to a strictly low-fat regimen . There are no clinical data on atherosclerosis reduction/regression , but animal models provide encouraging results . Role of NF-kappaB in the rescue of multiple myeloma cells from glucocorticoid-induced apoptosis by bcl-2 . The molecular mechanisms by which multiple myeloma ( MM ) cells evade glucocorticoid-induced apoptosis have not been delineated . Using a human IgAkappa MM cell line ( P24468 ) , we found that dexamethasone ( DB00514 ) -induced apoptosis is associated with decreased NF-kappaB DNA binding and kappaB-dependent transcription . Both nuclear p50:p50 and p50:p65 NF-kappaB complexes are detected in P24468 cells by supershift electrophoretic mobility shift assay ( EMSA ) . DB00514 -mediated inhibition of NF-kappaB DNA binding precedes a notable increase in annexin V binding , thereby indicating that diminished NF-kappaB activity is an early event in DB00514 -induced apoptosis . Overexpression of bcl-2 in P24468 cells prevents DB00514 -mediated repression of NF-kappaB activity and apoptosis . Sustained NF-kappaB DNA binding is also observed in two previously characterized DB00514 -resistant MM cell lines ( RPMI8226 and Q5SW96 -77 ) that express moderate levels of endogenous bcl-2 and P25963 proteins . In addition , enforced bcl-2 expression in P24468 cells did not prevent the augmentation of P25963 protein by DB00514 . We also noted a possible association between DB00514 -mediated downregulation of NF-kappaB in freshly obtained primary myeloma cells and the patients ' responsiveness to glucocorticoid-based chemotherapy . Collectively , our data suggest that the protective effects of bcl-2 in MM cells act upstream in the NF-kappaB activation-signaling pathway and the potential use of NF-kappaB as a biomarker in progressive MM . Loss of both phospholipid and triglyceride transfer activities of microsomal triglyceride transfer protein in abetalipoproteinemia . Mutations in microsomal triglyceride transfer protein ( P55157 ) cause abetalipoproteinemia ( P00519 ) , characterized by the absence of plasma apoB-containing lipoproteins . In this study , we characterized the effects of various P55157 missense mutations found in P00519 patients with respect to their expression , subcellular location , and interaction with protein disulfide isomerase ( P07237 ) . In addition , we characterized functional properties by analyzing phospholipid and triglyceride transfer activities and studied their ability to support apoB secretion . All the mutants colocalized with calnexin and interacted with P07237 . We found that R540H and N780Y , known to be deficient in triglyceride transfer activity , also lacked phospholipid transfer activity . Novel mutants S590I and G746E did not transfer triglycerides and phospholipids and did not assist in apoB secretion . In contrast , D384A displayed both triglyceride and phospholipid transfer activities and supported apoB secretion . These studies point out that P00519 is associated with the absence of both triglyceride and phospholipid transfer activities in P55157 . Interpretation of point-of-care INR results in patients treated with dabigatran . BACKGROUND : Point-of-care devices for measurement of the international normalized ratio ( INR ) are commonly used to monitor therapy and maintain therapeutic levels of anticoagulation in patients treated with vitamin K antagonists . DB06695 , a new oral , reversible direct thrombin inhibitor approved for stroke prevention in patients with atrial fibrillation does not require routine coagulation monitoring . However , case reports have identified falsely elevated point-of-care INR levels in patients treated with dabigatran using one of these devices ( Hemochron ) . This in vitro study was designed to verify this issue . METHODS : We compared INR levels in whole blood and plasma using a Hemochron Jr . Signature+ point-of-care device ( International Technidyne Corporation , Edison , NJ ) with routine laboratory monitoring , using blood from healthy volunteers that was spiked with increasing concentrations of dabigatran . RESULTS : P00734 time and INR levels were increased about 2- to 4-fold with the point-of-care device compared with laboratory measures across the plasma dabigatran concentration range 50-1400 ng/mL . At plasma concentrations of dabigatran likely to be observed in patients , at a dose of 150 mg twice daily ( 60-275 ng/mL ) , whole blood point-of-care INR values increased from 1.7 to 4.0 , versus 1.1 to 1.5 measured with the laboratory coagulometer . Similar differences in prothrombin time were observed in plasma samples . CONCLUSIONS : INR levels in patients taking dabigatran are substantially higher using a Hemochron Jr. point-of-care device compared with laboratory values . We discourage the use of these devices specifically , as well as the use of the INR in general , for measuring the anticoagulant effect of dabigatran . Inhibition of human brain and RBC acetylcholinesterase ( P22303 ) by heptylphysostigmine ( HPTL ) . Heptylphysostigmine ( HPTL ) , a derivative of the P22303 inhibitor physostigmine ( PHY ) , is under investigation as a therapeutic agent in Alzheimer 's disease . HPTL is active against human RBC P22303 both in vitro and in vivo . Activity of HPTL against human brain has not been documented . We have developed an in vitro assay system using particulate membrane fractions which permits comparison of inhibition and recovery kinetics of human RBC ( primarily globular dimer ) and brain ( primarily globular tetramer ) membrane-bound forms . Under these conditions the HPTLIC50 is similar for the two forms . RBC P22303 inhibition spontaneously reverses in 24 h , as occurs in vivo . In striking contrast , activity of inhibited brain enzyme does not recover on overnight incubation . DDVP-induced inhibition , but not HPTL inhibition , can be reversed by the oxime DB00733 . Some recovery of HPTL inhibition , but not to the level seen with RBC P22303 , occurs on addition of heat-stable fractions from serum or P04141 . Brain enzyme recovers rapidly from PHY in this system . Responses of brain and RBC P22303 to HPTL indicate that these forms are functionally as well as structurally distinct . Since brain inhibition apparently does not spontaneously reverse like RBC inhibition , peripheral measurements of patient responses should be assessed with caution during treatment with HPTL . Acute effects of acamprosate and MPEP on ethanol Drinking-in-the-Dark in male C57BL/6J mice . BACKGROUND : Recently , a simple procedure in mice , Drinking-in-the-Dark ( DID ) , was hypothesized to have value for medication development for human alcoholism . In DID , mice are offered intermittent , limited access to ethanol over a series of days during the dark phase that results in rapid drinking to intoxication in predisposed genotypes . METHODS : We measured the effects of acamprosate or MPEP , metabotropic glutamate 5 receptor ( P41594 ) antagonist , on intake of 20 % ethanol , plain tap water or 10 % sugar water using the DID procedure in male C57BL/6J mice . RESULTS : DB00659 ( 100 , 200 , 300 , or 400 mg/kg ) dose dependently decreased ethanol drinking with 300 mg/kg reducing ethanol intake by approximately 20 % without affecting intake of plain water or 10 % sugar water . MPEP ( 1 , 3 , 5 , 10 , 20 , or 40 mg/kg ) was more potent than acamprosate with 20 mg/kg reducing ethanol intake by approximately 20 % and for longer duration without affecting intake of plain water or 10 % sugar water . CONCLUSIONS : These results support the hypothesis that P41594 signaling plays a role in excessive ethanol intake in DID and suggest DID may have value for screening novel compounds that reduce overactive glutamate signaling for potential pharmaceutical treatment of excessive ethanol drinking behavior . DB04941 , an antisecretory antidiarrheal proanthocyanidin oligomer extracted from Croton lechleri , targets two distinct intestinal chloride channels . DB04941 , a purified proanthocyanidin oligomer extracted from the bark latex of Croton lechleri , is in clinical trials for secretory diarrheas of various etiologies . We investigated the antisecretory mechanism of crofelemer by determining its effect on the major apical membrane transport and signaling processes involved in intestinal fluid transport . Using cell lines and measurement procedures to isolate the effects on individual membrane transport proteins , crofelemer at 50 microM had little or no effect on the activity of epithelial Na(+) or K(+) channels or on DB02527 or calcium signaling . DB04941 inhibited the cystic fibrosis transmembrane regulator ( P13569 ) Cl(-) channel with maximum inhibition of approximately 60 % and an IC(50) approximately 7 microM . DB04941 action at an extracellular site on P13569 produced voltage-independent block with stabilization of the channel closed state . DB04941 did not affect the potency of glycine hydrazide or thiazolidinone P13569 inhibitors . DB04941 action resisted washout , with < 50 % reversal of P13569 inhibition after 4 h . DB04941 was also found to strongly inhibit the intestinal calcium-activated Cl(-) channel Q5XXA6 by a voltage-independent inhibition mechanism with maximum inhibition > 90 % and IC(50) approximately 6.5 microM . The dual inhibitory action of crofelemer on two structurally unrelated prosecretory intestinal Cl(-) channels may account for its intestinal antisecretory activity . Cyclooxygenase isozymes are expressed in human myeloma cells but not involved in anti-proliferative effect of cyclooxygenase inhibitors . Considering possible tumorigenic activity of cyclooxygenase ( P36551 ) isozymes in myeloma , we examined expression levels of P23219 and -2 in seven human myeloma cell lines ( Q5SW96 -77 , IM-9 , RPMI-8226 , HPC , HS-Sultan , TSPC-1 , and U-266 ) . As analyzed by reverse transcriptase-polymerase chain reaction ( RT-PCR ) , all the cell lines constitutively expressed P23219 , while P35354 levels markedly varied among different cell lines . Induction of P35354 by phorbol ester was observed in RPMI-8226 and HPC cells . In contrast , P35354 was constitutively expressed in Q5SW96 -77 and IM-9 cells . Moreover , the high expression level of P35354 protein in Q5SW96 -77 cells was verified by Western blotting . Intact cells of Q5SW96 -77 converted 14C-labeled arachidonic acid to prostaglandin E2 , F2alpha , and D2 , and this activity was dose-dependently inhibited by selective P35354 inhibitors ( SC-58125 and NS-398 ) , a non-selective P36551 inhibitor ( indomethacin ) , and relatively high concentrations of a selective P23219 inhibitor ( SC-560 ) . These P36551 inhibitors also suppressed the proliferation of Q5SW96 -77 cells , but significant suppression was seen only at 100 microM , a much higher concentration than those sufficient for the P36551 inhibition . Moreover , proliferation of the myeloma cells lacking P35354 was also suppressed by 100 microM of SC-58125 . These results suggested that the anti-proliferative effect of the P36551 inhibitors is independent of the inhibition of P35354 . Gateways to clinical trials . Gateways to Clinical Trials is a guide to the most recent clinical trials in current literature and congresses . The data in the following tables has been retrieved from the Clinical Studies Knowledge Area of Prous Science Integrity(R) , the drug discovery and development portal , http://integrity.prous.com . This issue focuses on the following selection of drugs : 3,4-DAP ; DB00718 , Q16586 -10-0101 , alefacept , alemtuzumab , alosetron hydrochloride , ALT-711 , aprepitant , atazanavir sulfate , atlizumab , atvogen ; DB00188 ; P11597 vaccine , clevudine , crofelemer ; P22760 : P0C6A0 , darbepoetin alfa , decitabine , drotrecogin alfa ( activated ) , DX-9065a ; E-7010 , edodekin alfa , emivirine , emtricitabine , entecavir , erlosamide , erlotinib hydrochloride , everolimus , exenatide ; DB00569 , frovatriptan , fulvestrant ; DB00056 , gestodene ; DB04865 , human insulin ; Imatinib mesylate , indiplon , indium 111 ( 111In ) ibritumomab tiuxetan , inhaled insulin , insulin detemir , insulin glargine , ivabradine hydrochloride ; DB06792 , lapatinib , O43766 -34475 , levetiracetam , liraglutide , lumiracoxib ; Maxacalcitol , melagatran , micafungin sodium ; DB00108 , NSC-640488 ; Oblimersen sodium ; DB08439 sodium , PEG-filgrastim , peginterferon alfa-2(a) , peginterferon alfa-2b , pexelizumab , pimecrolimus , pleconaril , pramlintide acetate , pregabalin , prucalopride ; DB00025 -PFM , Ranelic acid distrontium salt , ranolazine , rDNA insulin , recombinant human soluble thrombomodulin , rhGM- P04141 , roxifiban acetate , RSD-1235 , rubitecan , ruboxistaurin mesilate hydrate ; SC-51 , squalamine ; DB01079 maleate , telbivudine , tesaglitazar , testosterone gel , tezosentan disodium , tipranavir ; DB04879 succinate ; DB04898 ; Yttrium 90 ( 90Y ) ibritumomab tiuxetan ; DB00399 monohydrate . Human DB03501 4 ' epimerase ( Q14376 ) gene and identification of five missense mutations in patients with epimerase-deficiency galactosemia . The galactosemias are a series of three inborn errors of metabolism caused by deficiency of any one of the three human galactose-metabolic enzymes : galactokinase ( P51570 ) , galactose-1-phosphate uridyl transferase ( P07902 ) , and DB03501 4 ' epimerase ( Q14376 ) . We report here the characterization of the entire coding sequence of the Q14376 gene and screening for mutations in epimerase-deficient individuals . The human Q14376 gene is about 4 kb in size and is divided into 11 exons on chromosome band 1p36 . We have identified five mutations in the Q14376 gene of epimerase-deficient galactosemia patients . The patients were either homozygotes or compound heterozygotes for mutations . These results confirm that epimerase-deficiency galactosemia is the result of missense mutations in the Q14376 gene and indicate that the disease is characterized by extensive allelic heterogeneity . Discovery and evaluation of 3-phenyl-1H-5-pyrazolylamine-based derivatives as potent , selective and efficacious inhibitors of P07333 -like tyrosine kinase-3 ( P36888 ) . Preclinical investigations and early clinical trial studies suggest that P36888 inhibitors offer a viable therapy for acute myeloid leukemia . However , early clinical data for direct P36888 inhibitors provided only modest results because of the failure to fully inhibit P36888 . We have designed and synthesized a novel class of 3-phenyl-1H-5-pyrazolylamine-derived compounds as P36888 inhibitors which exhibit potent P36888 inhibition and high selectivity toward different receptor tyrosine kinases . The structure-activity relationships led to the discovery of two series of P36888 inhibitors , and some potent compounds within these two series exhibited comparable potency to P36888 inhibitors sorafenib ( 3 ) and DB06080 ( 4 ) in both wt- P36888 enzyme inhibition and P36888 -ITD inhibition on cell growth ( MOLM-13 and MV4;11 cells ) . In particular , the selected compound 12a exhibited the ability to regress tumors in mouse xenograft models using MOLM-13 and MV4;11 cells . Renal changes induced by a cyclooxygenase-2 inhibitor during normal and low sodium intake . P35354 ( P35354 ) has been identified in renal tissues under normal conditions , with its expression enhanced during sodium restriction . To evaluate the role of P35354 -derived metabolites in the regulation of renal function , we infused a selective inhibitor ( nimesulide ) in anesthetized dogs with normal or low sodium intake . The renal effects elicited by nimesulide and a non-isozyme-specific inhibitor ( meclofenamate ) were compared during normal sodium intake . In ex vivo assays , meclofenamate , but not nimesulide , prevented the platelet aggregation elicited by arachidonic acid . During normal sodium intake , nimesulide infusion ( n=6 ) had no effects on arterial pressure or renal hemodynamics but did reduce urinary sodium excretion , urine flow rate , and fractional lithium excretion . In contrast , nimesulide administration increased arterial pressure and decreased renal blood flow , urine flow rate , and fractional lithium excretion during low sodium intake ( n=6 ) . P35354 inhibition reduced urinary prostaglandin E(2) excretion in both groups but did not modify plasma renin activity in dogs with low ( 8.1+/-1.1 ng angiotensin I. mL(-1). h(-1) ) or normal ( 1.8+/-0.4 ng angiotensin I. mL(-1). h(-1) ) sodium intake . DB00939 infusion in dogs with normal sodium intake ( n=8 ) induced a greater renal hemodynamic effect than nimesulide infusion . These results suggest that P35354 -derived metabolites ( 1 ) are involved in the regulation of sodium excretion in dogs with normal sodium intake , ( 2 ) play an important role in the regulation of renal hemodynamic and excretory function in dogs with low sodium intake , and ( 3 ) are not involved in the maintenance of the high renin levels during a long-term decrease in sodium intake . Islet-selectivity of G-protein coupled receptor ligands evaluated for PET imaging of pancreatic β-cell mass . A critical unmet need exists for methods to quantitatively measure endogenous pancreatic β-cell mass ( Q02223 ) for the clinical evaluation of therapies to prevent or reverse loss of Q02223 and diabetes progression . Our objective was to identify G-protein coupled receptors ( GPCRs ) that are expressed with a high degree of specificity to islet β-cells for receptor-targeted imaging of Q02223 . GPCRs enriched in pancreatic islets relative to pancreas acinar and hepatic tissue were identified using a database screen . Islet-specific expression was confirmed by human pancreas immunohistochemistry ( IHC ) . In vitro selectivity assessment was determined from the binding and uptake of radiolabeled ligands to the rat insulinoma P01308 -1 832/13 cell line and isolated rat islets relative to the exocrine pancreas cell-type , PANC-1 . Tail-vein injections of radioligands into rats were used to determine favorable image criteria of in vivo biodistribution to the pancreas relative to other internal organs ( i.e. , liver , spleen , stomach , and lungs ) . Database and IHC screening identified four candidate receptors for further in vitro and in vivo evaluation for PET imaging of Q02223 : prokineticin-1 receptor ( PK-1R ) , metabotropic glutamate receptor type-5 ( P41594 ) , neuropeptide Y-2 receptor ( P01303 -2R ) , and glucagon-like peptide 1 receptor ( P43220 ) . In vitro specificity ratios gave the following receptor rank order : PK-1R > P43220 > P01303 -2R > P41594 . The biodistribution rank order of selectivity to the pancreas was found to be PK-1R > VMAT2∼GLP-1R > P41594 . Favorable islet selectivity and biodistribution characteristics suggest several GPCRs as potential targets for PET imaging of pancreatic Q02223 .

MH_train_1342

MH_train_1342

MH_train_1342

interacts_with DB00266?

multiple_choice

Establishment by adriamycin exposure of multidrug-resistant rat ascites hepatoma AH130 cells showing low P15559 activity and high cross resistance to mitomycins . A resistant subline ( AH130/5A ) selected from rat hepatoma AH130 cells after exposure to adriamycin ( P35318 ) showed remarkable resistance to multiple antitumor drugs , including mitomycin C ( DB00305 ) and porfiromycin ( PFM ) . PFM , vinblastine ( DB00570 ) , and P35318 accumulated in AH130/5A far less than in the parent AH130 ( AH130/P ) cells . AH130/5A cells showed overexpression of P-glycoprotein ( A6NDG6 ) , an increase in glutathione S-transferase activity , and a decrease in P15559 and glutathione peroxidase activity . The resistance to DB00305 and DB00570 of AH130/5A cells was partly reversed by H-87 , an inhibitor of A6NDG6 . Buthionine sulfoximine , an inhibitor of glutathione synthase , did not affect the action of DB00305 . tert-Butylhydroquinone induced P15559 activity , increased PFM uptake , and enhanced the growth-inhibitory action of DB00305 in AH130/5A cells . DB00266 , an inhibitor of P15559 , decreased PFM uptake and reduced the growth-inhibitory action of DB00305 in AH130/P cells . These results indicated that the adriamycin treatment of hepatoma cells caused multifactorial multidrug resistance involving a decrease in P15559 activity . DB05311 ( DX-88 ) , a plasma kallikrein inhibitor for the treatment of hereditary angioedema and the prevention of blood loss in on-pump cardiothoracic surgery . BACKGROUND : P03952 plays a major role in the contact ( kallikrein-kinin ) cascade producing bradykinin . Bradykinin is a vasodilator , which increases vascular permeability , activates inflammation and produces pain . P03952 is also crosslinked to the coagulation system and the complement cascade . OBJECTIVE : DB05311 ( DX-88 ) is a potent and specific inhibitor of plasma kallikrein . DB05311 is a recombinantly produced and engineered small protein based on the first Kunitz domain of human tissue factor pathway inhibitor . It was identified through phage display technology . METHODS : The search terms ' ecallantide ' , ' DX-88 ' and ' hereditary angioedema ' were entered into Pubmed/Medline , ClinicalTrials and Google . RESULTS/CONCLUSION : At present , the drug is being studied for two major indications . First , the results for the treatment of hereditary angioedema are promising . Second , a prospective randomised multi-centre trial for the reduction of blood loss during on-pump cardiothoracic surgery will be terminated in October 2008 . Development of water soluble derivatives of cis-3 , 4 ' , 5-trimethoxy-3'-aminostilbene for optimization and use in cancer therapy . DB01394 site tubulin inhibitors are currently developed as vascular disrupting agents ( VDAs ) . However , they were found to have cardiotoxicity in clinical trials . To overcome the problem , we developed a stilbene derivative , cis-3 , 4 ' , 5-trimethoxy-3'-aminostilbene ( stilbene 5c ) , which is highly potent and has no bone marrow and cardiac toxicity in mice . Here we attempt to optimize stilbene 5c using computer-based drug design and synthesize derivatives with benzimidazole or indole group . Biological evaluation showed that they are weaker than stilbene 5c without better water solubility . Alternative approach was thus adopted to make prodrugs of stilbene 5c . A water-soluble prodrug PD7 was synthesized by addition of a morpholino group with carbamate linkage to the amino group of stilbene 5c . In vitro studies show that PD7 induces mitotic arrest and disrupts microtubule similar to stilbene 5c . The cell signaling events in Cdc2 , p53 , Akt , and aurora kinase are similar in cells treated with stilbene 5c , P22748 or PD7 , suggesting that they share the same mechanism . Although PD7 is less effective than stilbene 5c in vitro , the biological activity of PD7 as a single agent is similar to that of stilbene 5c . Combination of PD7 with P15692 inhibitor bevacizumab significantly enhances the therapeutic efficacy of PD7 in mouse xenograft model . These data suggest that PD7 could be a good candidate for further pre-clinical and clinical development as a new VDA for cancer therapy . A phase-1 trial of bexarotene and denileukin diftitox in patients with relapsed or refractory cutaneous T-cell lymphoma . DB00004 , a genetically engineered fusion protein combining the enzymatically active domains of diphtheria toxin and the full-length sequence for interleukin-2 ( P60568 ) , efficiently targets lymphoma cells expressing the high-affinity P60568 receptor ( IL-2R ) consisting of the alpha/p55/CD25 , beta/p75/CD122 , and gamma/ P31785 /CD132 chains . In vitro studies demonstrated that the retinoid X receptor ( RXR ) retinoid , bexarotene , at biologically relevant concentrations of 10(-6) M to 10(-8) M , upregulated both the p55 and p75 subunits of the IL-2R and enhanced 5- to 10-fold the susceptibility of T-cell leukemia cells to denileukin diftitox . To determine whether this biomodulatory effect could be recapitulated in vivo , we treated 14 patients with relapsed or refractory cutaneous T-cell lymphoma with escalating doses of bexarotene ( 75 mg/day-300 mg/day ) and denileukin diftitox ( 18 mcg/kg per day x 3 days every 21 days ) in a phase 1 trial . Overall response was 67 % ( 4 complete responses , 4 partial responses ) . Modulation of IL-2R expression was observed at or above a bexarotene dose of 150 mg/day . Four patients experienced grade 2 or 3 leukopenia , and 2 had grade 4 lymphopenia . Our results demonstrate that the combination of denileukin diftitox and bexarotene is well tolerated and that even low doses ( 150 mg/day ) of bexarotene are capable of in vivo upregulation of CD25 expression on circulating leukemia cells . A synthetic dl-nordihydroguaiaretic acid ( Nordy ) , inhibits angiogenesis , invasion and proliferation of glioma stem cells within a zebrafish xenotransplantation model . The zebrafish ( Danio rerio ) and their transparent embryos represent a promising model system in cancer research . Compared with other vertebrate model systems , we had previously shown that the zebrafish model provides many advantages over mouse or chicken models to study tumor invasion , angiogenesis , and tumorigenesis . In this study , we systematically investigated the biological features of glioma stem cells ( GSCs ) in a zebrafish model , such as tumor angiogenesis , invasion , and proliferation . We demonstrated that several verified anti-angiogenic agents inhibited angiogenesis that was induced by xenografted-GSCs . We next evaluated the effects of a synthetic dl-nordihydroguaiaretic acid compound ( dl-NDGA or " Nordy " ) , which revealed anti-tumor activity against human GSCs in vitro by establishing parameters through studying its ability to suppress angiogenesis , tumor invasion , and proliferation . Furthermore , our results indicated that Nordy might inhibit GSCs invasion and proliferation through regulation of the arachidonate P09917 ( Alox-5 ) pathway . Moreover , the combination of Nordy and a P15692 inhibitor exhibited an enhanced ability to suppress angiogenesis that was induced by GSCs . By contrast , even following treatment with 50 µM Nordy , there was no discernible effect on zebrafish embryonic development . Together , these results suggested efficacy and safety of using Nordy in vivo , and further demonstrated that this model should be suitable for studying GSCs and anti- P56915 drug evaluation . 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 . Retinal gene expression and Müller cell responses after branch retinal vein occlusion in the rat . PURPOSE : In a rat model of branch retinal vein occlusion ( BRVO ) , changes in gene expression of factors implicated in the development of retinal edema and alterations in the properties of Müller cells were determined . METHODS : In adult Long-Evans rats , BRVO was induced by laser photocoagulation of retinal veins ; untreated eyes served as controls . The mRNA levels of after factors were determined with real-time RT-PCR in the neural retina and retinal pigment epithelium after 1 and 3 days of BRVO : P15692 , pigment epithelium-derived factor ( P36955 ) , tissue factor , prothrombin , the potassium channel Kir4.1 , and aquaporins 1 and 4 . DB01345 currents were recorded in isolated Müller cells , and cellular swelling was assessed in retinal slices . RESULTS : In the neural retina , the expression of P15692 was upregulated within 1 day of BRVO and returned to the control level after 3 days . P36955 was upregulated in the neuroretina and retinal pigment epithelium after 3 days of BRVO . P00734 , Kir4.1 , and both aquaporins were downregulated in the neuroretina . After BRVO , Müller cells displayed a decrease in their potassium currents and an altered distribution of Kir4.1 protein , an increase in the size of their somata , and cellular swelling under hypoosmotic stress that was not observed in control tissues . CONCLUSIONS : BRVO results in a rapid transient increase in the expression of P15692 and a delayed increase in the expression of P36955 . The downregulation of Kir4.1 and aquaporins , the mislocation of Kir4.1 protein , and the osmotic swelling of Müller cells may contribute to the development of edema and neuronal degeneration . Pharmacological profile of astemizole-derived compounds at the histamine H1 and H4 receptor -- H1/H4 receptor selectivity . DB00637 , a P35367 antagonist shows high affinity to the histamine H1 receptor but only a moderate affinity to the histamine H4 receptor . This study aims to modify the astemizole to keep high affinity to the histamine H1 receptor and to increase affinity to the histamine H4 receptor . Therefore , 13 astemizole-derived compounds and astemizole-JNJ7777120-derived hybrid compounds were synthesized and pharmacologically characterized at the histamine H1 and H4 receptors . The new compounds show affinity to the histamine H1 receptor in the pK i range from 5.3 to 8.8 , whereas the affinity of these compounds to the histamine H4 receptor was surprisingly rather low ( pK i from 4.4 to 5.6 ) . Three representative compounds were docked into the histamine H1 receptor and molecular dynamic studies were performed to explain the binding mode and the experimental results on a molecular level . Furthermore , taking into account the binding mode of compounds with high affinity to the histamine H4 receptor , a H1/H4-pharmacophore hypothesis was developed . DB04873 ( SB 207266 ) , a selective Q13639 receptor antagonist , reduces serotonin potentiation of neurally-mediated contractile responses of human detrusor muscle . The aim of this study is to evaluate the potency of piboserod ( SB 207266 ) , a selective 5-HT(4) receptor antagonist , at inhibiting the 5-HT(4)-mediated potentiating effect of serotonin ( 5-HT ) on the neurally-mediated contractile responses of human detrusor strips to electrical field stimulations ( O43281 ) . Strips of human detrusor muscle were mounted in Krebs-HEPES buffer under a resting tension of 500 mg and O43281 ( 20 Hz , 1 ms duration at 300 mA for 5 s ) was applied continuously at 1 min intervals . After stabilization of the O43281 -induced contractions , concentration-response curves to 5-HT ( 0.1 nM-100 microM ) were constructed in the absence or presence of 1 or 100 nM of piboserod . The experiments were performed in the presence of methysergide ( 1 microM ) and ondansetron ( 3 microM ) to block 5HT(1)/5HT(2) and 5-HT(3) receptors , respectively . 5-HT potentiated the contractile responses to O43281 of human bladder strips in a concentration-dependent manner , with a maximum mean of 60.0+/-19.9 % of the basal O43281 -evoked contractions . DB04873 did not modify the basal contractions but concentration-dependently antagonized the ability of 5-HT to enhance bladder strip contractions to O43281 . In presence of 1 and 100 nM of piboserod , the maximal 5-HT-induced potentiations were reduced to 45.0+/-7.9 and 38.7+/-8.7 % , respectively . A mean apparent antagonist dissociation constant value ( K(B) ) of 0.56+/-0.09 nM was determined . These data show the ability of piboserod to antagonize with high potency the enhancing properties of 5-HT on neurally-mediated contractions of isolated human bladder strips . Therefore , the 5-HT(4) receptor might represent an attractive pharmacological target for the treatment of overactive bladder . Leukotriene C4 production during hypoxic pulmonary vasoconstriction in isolated rat lungs . Leukotriene inhibitors preferentially inhibit hypoxic pulmonary vasoconstriction in isolated rat lungs . If lipoxygenase products are involved in the hypoxic pressor response they might be produced during acute alveolar hypoxia and a leukotriene inhibitor should block both the vasoconstriction and leukotriene production that occurs in response to hypoxia . We investigated in isolated blood perfused rat lungs whether leukotriene C4 ( LTC4 ) could be recovered from whole lung lavage fluid during ongoing hypoxic vasoconstriction . Lung lavage from individual rats had slow reacting substance ( SRS ) -like myotropic activity by guinea pig ileum bioassay . Pooled lavage ( 10 lungs ) as analyzed by reverse phase high performance liquid chromatography had an ultraviolet absorbing component at the retention time for LTC4 . At radioimmunoassay , and SRS myotropic activity by bioassay . LTC4 was not found during normoxic ventilation , during normoxic ventilation after a hypoxic pressor response , or during vasoconstriction elicited by DB00761 . DB00711 citrate , a leukotriene synthesis blocker , concomitantly inhibited the hypoxic vasoconstriction and LTC4 production . Thus P09917 products may play a role in the sequence of events leading to hypoxic pulmonary vasoconstriction . Targeting eIF4GI translation initiation factor affords an attractive therapeutic strategy in multiple myeloma . BACKGROUND : Deregulation of protein synthesis is integral to the malignant phenotype and translation initiation is the rate limiting stage . Therefore , eIF4F translation initiation complex components are attractive therapeutic targets . METHODS : Protein lysates of myeloma cells ( cell lines/patients ' bone marrow samples ) untreated/treated with bevacizumab were assayed for eIF4GI expression , regulation ( P15559 /proteosome dependent fragmentation ) ( WB , DB00266 , qPCR ) and targets (WB). eIF4GI was inhibited by knockdown and 4EGI-1 . Cells were tested for viability ( ELISA ) , death ( FACS ) and eIF4GI targets ( WB ) . RESULTS : Previously , we have shown that manipulation of P15692 in myeloma cells attenuated P06730 dependent translation initiation . Here we assessed the significance of eIF4GI to MM cells . We demonstrated increased expression of eIF4GI in myeloma cells and its attenuation upon P15692 inhibition attributed to elevated P15559 /proteasome dependent fragmentation and diminished mRNA levels . Knockdown of eIF4GI was deleterious to myeloma cells phenotype and expression of specific molecular targets ( Q99717 /ERα/HIF1α/c-Myc ) . Finally , we showed that the small molecule 4EGI-1 inhibits eIF4GI and causes a reduction in expression of its molecular targets in myeloma . CONCLUSION : Our findings substantiate that translation initiation of particular targets in MM is contingent on the function of eIF4GI , critical to cell phenotype , and mark it as a viable target for pharmacological intervention . Structures of murine carbonic anhydrase IV and human carbonic anhydrase II complexed with brinzolamide : molecular basis of isozyme-drug discrimination . P22748 ( CAIV ) is a membrane-associated enzyme anchored to plasma membrane surfaces by a phosphatidylinositol glycan linkage . We have determined the 2.8-angstroms resolution crystal structure of a truncated , soluble form of recombinant murine CAIV . We have also determined the structure of its complex with a drug used for glaucoma therapy , the sulfonamide inhibitor brinzolamide ( DB01194 ) . The overall structure of murine CAIV is generally similar to that of human CAIV ; however , some local structural differences are found in the active site resulting from amino acid sequence differences in the " 130 's segment " and the residue-63 loop ( these may affect the nearby catalytic proton shuttle , DB00117 -64 ) . Similar to human CAIV , the C-terminus of murine CAIV is surrounded by a substantial electropositive surface potential that may stabilize the interaction with the phospholipid membrane . Binding interactions observed for brinzolamide rationalize the generally weaker affinity of inhibitors used in glaucoma therapy toward CAIV compared with CAII . Antidepressant properties of the Q13639 receptor partial agonist , SL65.0155 : behavioral and neurochemical studies in rats . This study was undertaken to investigate the potential antidepressant-like properties of SL65.0155 , a serotonin 5-HT(4) receptor partial agonist , in male rats of the Wistar strain tested in the forced swim test ( P19883 ) , an experimental model widely used to assess antidepressant-like activity . The expression of hippocampal neurotrophic factors , such as the brain-derived neurotrophic factor ( P23560 ) , the phosphorilated DB02527 response element-binding protein ( p-CREB ) , the B cell lymphoma-2 ( Bcl-2 ) , the Bax and the vascular endothelium growth factor ( P15692 ) were also evaluated by Western Blot analysis . Different groups of rats received intraperitoneally ( i.p. ) injections of SL65.0155 ( 0.1 , 0.5 and 1 mg/kg ) , clomipramine ( 50 mg/kg ) , citalopram ( 15 mg/kg ) or vehicle , respectively , 24 , 5 and 1 h prior to the P19883 . Compared to the control group , SL65.0155 ( 0.5 and 1 mg/kg ) , clomipramine or citalopram injected animals showed an increased swimming and climbing behavior and reduced immobility time in the P19883 . Interestingly , this effect was not due to changes in the locomotor activity since all treated groups failed to show any change in motor ability as assessed in the open field test . Western blot analysis of hippocampal homogenates showed an enhancement of p-CREB , P23560 Bcl-2 and P15692 protein levels in SL65.0155 treated groups , but not in citalopram or clomipramine treated groups , used here as positive control . No change was found in Bax expression in any treated group . These findings give further support to the hypothesis that the stimulation of serotonin 5-HT(4) receptors may be a therapeutic target for depression . Genome-wide association study on plasma levels of midregional-proadrenomedullin and C-terminal-pro-endothelin-1 . P05305 ( ET-1 ) and adrenomedullin ( P35318 ) are circulating vasoactive peptides involved in vascular homeostasis and endothelial function . Elevated levels of plasma ET-1 and P35318 , and their biologically stable surrogates , C-terminal-pro-endothelin-1 ( CT-pro-ET-1 ) and midregional proadrenomedullin ( MR-pro- P35318 ) , are predictors of cardiac death and heart failure . We studied the association of common genetic variation with MR-pro- P35318 and CT-pro-ET-1 by genome-wide association analyses in 3444 participants of European ancestry . We performed follow-up genotyping of single nucleotide polymorphisms ( SNPs ) that showed suggestive or significant association in the discovery stage in additional 3230 participants . The minor variants in P03952 ( rs4253238 ) and P00748 ( rs2731672 ) , both part of the kallikrein-kinin system , were associated with higher MR-pro- P35318 ( P=4.46E-52 and P=5.90E-24 , respectively ) and higher CT-pro-ET-1 levels ( P=1.23E-122 and P=1.26E-67 , respectively ) . Epistasis analyses showed a significant interaction between the sentinel SNP of P00748 and P03952 for both traits . In addition , a variant near the P35318 gene ( rs2957692 ) was associated with MR-pro- P35318 ( P=1.05E-12 ) and a variant in P10153 -1 ( rs5370 ) was associated with CT-pro-ET-1 ( P=1.49E-27 ) . The total phenotypic variation explained by the genetic variants was 7.2 % for MR-pro- P35318 and 14.6 % for CT-pro-ET-1 . P03952 encodes plasma kallikrein , a proteolytic enzyme known to cleave high-molecular-weight kininogen to bradykinin and prorenin to renin . We cloned the precursors of P35318 and ET-1 and demonstrated that purified plasma kallikrein can cleave these recombinant proteins into multiple smaller peptides . The discovery of genetic variants in the kallikrein-kinin system and in the genes encoding pre-pro-ET-1 and pre-pro- P35318 provides novel insights into the (co-)regulation of these vasoactive peptides in the vascular system . DB00428 -nicotinamide-induced diabetes in the rat . Characteristics of the experimental model . Administration of both streptozotocin ( Q11206 ) and nicotinamide ( NA ) has been proposed to induce experimental diabetes in the rat . Q11206 is well known to cause pancreatic B-cell damage , whereas NA is administered to rats to partially protect insulin-secreting cells against Q11206 . Q11206 is transported into B-cells via the glucose transporter P11168 and causes DNA damage leading to increased activity of poly(ADP-ribose) polymerase ( P09874 ) to repair DNA . However , exaggerated activity of this enzyme results in depletion of intracellular NAD(+) and DB00171 , and the insulin-secreting cells undergo necrosis . The protective action of NA is due to the inhibition of P09874 activity . NA inhibits this enzyme , preventing depletion of NAD(+) and DB00171 in cells exposed to Q11206 . Moreover , NA serves as a precursor of NAD(+) and thereby additionally increases intracellular NAD(+) levels . The severity of diabetes in experimental rats strongly depends on the doses of Q11206 and NA given to these animals . Therefore , in diabetic rats , blood glucose may be changed in a broad range -- from slight hyperglycemia to substantial hyperglycemia compared with control animals . Similarly , blood insulin may be only slightly decreased or substantial hypoinsulinemia may be induced . In vitro studies demonstrated that the insulin-secretory response to glucose is attenuated in Q11206 -NA-induced diabetic rats compared with control animals . This is due to reduced B-cell mass as well as metabolic defects in the insulin-secreting cells . Results of numerous experiments have demonstrated that this model of diabetes is useful in studies of different aspects of diabetes . Interpretation of point-of-care INR results in patients treated with dabigatran . BACKGROUND : Point-of-care devices for measurement of the international normalized ratio ( INR ) are commonly used to monitor therapy and maintain therapeutic levels of anticoagulation in patients treated with vitamin K antagonists . DB06695 , a new oral , reversible direct thrombin inhibitor approved for stroke prevention in patients with atrial fibrillation does not require routine coagulation monitoring . However , case reports have identified falsely elevated point-of-care INR levels in patients treated with dabigatran using one of these devices ( Hemochron ) . This in vitro study was designed to verify this issue . METHODS : We compared INR levels in whole blood and plasma using a Hemochron Jr . Signature+ point-of-care device ( International Technidyne Corporation , Edison , NJ ) with routine laboratory monitoring , using blood from healthy volunteers that was spiked with increasing concentrations of dabigatran . RESULTS : P00734 time and INR levels were increased about 2- to 4-fold with the point-of-care device compared with laboratory measures across the plasma dabigatran concentration range 50-1400 ng/mL . At plasma concentrations of dabigatran likely to be observed in patients , at a dose of 150 mg twice daily ( 60-275 ng/mL ) , whole blood point-of-care INR values increased from 1.7 to 4.0 , versus 1.1 to 1.5 measured with the laboratory coagulometer . Similar differences in prothrombin time were observed in plasma samples . CONCLUSIONS : INR levels in patients taking dabigatran are substantially higher using a Hemochron Jr. point-of-care device compared with laboratory values . We discourage the use of these devices specifically , as well as the use of the INR in general , for measuring the anticoagulant effect of dabigatran . A general method for the synthesis of aryl [11C]methylsulfones : potential PET probes for imaging cyclooxygenase-2 expression . A general one-pot method has been developed for the conversion of an aryl thiol moiety masked as the butyrate ester to the corresponding 11C-labeled methylsulfone group . The potential of this methodology has been demonstrated by the successful radiosynthesis of carbon-11 analogues of several highly selective cyclooxygenase-2 ( P35354 ) inhibitors such as DB00533 , DB01628 , and 3-(4-methylsulfonylphenyl)-4-phenyl-5-trifluoromethyl isoxazole in high yield . The chemical and radiochemical purities obtained for the 11C-labeled P35354 inhibitors are > 99 % with a specific activity > 1000 Ci/mmol . Gene expression by PBMC in primary sclerosing cholangitis : evidence for dysregulation of immune mediated genes . Primary sclerosing cholangitis ( PSC ) is a chronic disease of the bile ducts characterized by an inflammatory infiltrate and obliterative fibrosis . The precise role of the immune system in the pathogenesis of PSC remains unknown . We used RNA microarray analysis to identify immune-related genes and pathways that are differentially expressed in PSC . Messenger RNA ( mRNA ) from peripheral blood mononuclear cells ( PBMC ) was isolated from both patients with PSC and age and sex matched healthy controls . Samples from 5 PSC patients and 5 controls were analyzed by microarray and based upon rigorous statistical analysis of the data , relevant genes were chosen for confirmation by RT-PCR in 10 PSC patients and 10 controls . Using unsupervised hierarchical clustering , gene expression in PSC was statistically different from our control population . Interestingly , genes within the P60568 receptor beta , P05231 and Q96HU1 Kinase pathways were found to be differently expressed in patients with PSC compared to controls . Further , individual genes , P01375 induced protein 6 ( TNFaip6 ) and membrane-spanning 4-domains , subfamily A ( ms4a ) were found to be upregulated in PSC while similar to Q99717 ( Q99717 ) was downregulated . In conclusion , several immune-related pathways and genes were differentially expressed in PSC compared to control patients , giving further evidence that this disease is systemic and immune-mediated . Dynamic stereochemistry of Topiramate ( anticonvulsant drug ) in solution : theoretical approaches and experimental validation . Topiramate , an antiepileptic drug , was synthesized with an improved protocol and identified by (1)H NMR , (13)C NMR , (1)H-(1)H COSY , HMQC and HMBC spectrum . In parallel , density functional theory ( DFT ) using B3LYP functional and split-valance 6-311++G** basis set has been used to optimize the structures and conformers of Topiramate . Also experimental and theoretical methods have been used to correlate the dependencies of (1)J and (2)J involving (1)H and (13)C on the C1- P06681 ( ω ) and C1-O1 ( θ ) torsion angles in the glycosidic part of Topiramate . New Karplus equations are proposed to assist in the structural interpretation of these couplings . Importantly , due to the sensitivity of some couplings , most notably (2)J( P35367 ,H1S) , (2)J( P06681 , P35367 ) and (2)J( P06681 ,H1S) values depend on both C-C ( ω ) and C-O ( θ ) torsion angles . Analyses of experimental coupling constants for protons on the pyranose ring of Topiramate indicate a twist boat structure for Topiramate in solution . In all calculations solvent effects were considered using a polarized continuum model ( PCM ) . DMPPQA , a novel angiogenesis inhibitor , induces apoptosis in human colon cancer HCT-116 cells and HUVECs . Cytotoxic activity of 5,7-dimethoxy-2-phenyl-N-propylquinolin-4-amine ( DMPPQA ) was investigated in human colon cancer cells HCT-116 and umbilical vein endothelial cell line HUVEC . The IC(50) of DMPPQA on HCT-116 and HUVEC cells were respectively 1.26 and 7.43 µM after 72 h treatment . DMPPQA inhibited the growth of HCT-116 and HUVEC cells in concentration- and time-dependent manners . Typical morphological changes of apoptotic body formation were seen after DMPPQA with Hoechst 33258 staining . DB00828 analysis showed that DMPPQA induced apoptosis , mitochondrial membrane potential loss ( ΔΨm ) and increase in the production of intracellular reactive oxygen species ( ROS ) of HCT-116 cells . After treating with DMPPQA , apoptosis-related protein expression of Bax , cytochrome c , caspase-9 , caspase-3 , P09874 and P04637 increased and Bcl-2 protein expression decreased . DMPPQA treatment of HUVECs reduced cell migration and microcapillary tube formation in a Matrigel matrix . It also decreased P15692 protein expression . Thus DMPPQA acts as an angiogenesis inhibitor and induces cell apoptosis by a caspase-dependent mitochondrial pathway . Pivotal role of the P06681 domain of the Smurf1 ubiquitin ligase in substrate selection . The P06681 -WW-HECT-type ubiquitin ligases Smurf1 and Smurf2 play a critical role in embryogenesis and adult bone homeostasis via regulation of bone morphogenetic protein , Wnt , and RhoA signaling pathways . The intramolecular interaction between P06681 and HECT domains autoinhibits the ligase activity of Smurf2 . However , the role of the Smurf1 P06681 domain remains elusive . Here , we show that the P06681 -HECT autoinhibition mechanism is not observed in Smurf1 , and instead its P06681 domain functions in substrate selection . The Smurf1 P06681 domain exerts a key role in localization to the plasma membrane and endows Smurf1 with differential activity toward RhoA versus Q99717 and Runx2 . Crystal structure analysis reveals that the Smurf1 P06681 domain possesses a typical anti-parallel β-sandwich fold . Examination of the sulfate-binding site analysis reveals two key lysine residues , Lys-28 and Lys-85 , within the P06681 domain that are important for Smurf1 localization at the plasma membrane , regulation on cell migration , and robust ligase activity toward RhoA , which further supports a Ca(2+)-independent localization mechanism for Smurf1 . These findings demonstrate a previously unidentified role of the Smurf1 P06681 domain in substrate selection and cellular localization .

MH_train_1343

MH_train_1343

MH_train_1343

interacts_with DB00222?

multiple_choice

Intracellular targets of cyclin-dependent kinase inhibitors : identification by affinity chromatography using immobilised inhibitors . BACKGROUND : Chemical inhibitors of cyclin-dependent kinases ( CDKs ) have great therapeutic potential against various proliferative and neurodegenerative disorders . DB02116 , a 2,6,9-trisubstituted purine , has been optimized for activity against P06493 /cyclin B by combinatorial and medicinal chemistry efforts to yield the purvalanol inhibitors . Although many studies support the action of purvalanols against CDKs , the actual intracellular targets of 2,6 , 9-trisubstituted purines remain unverified . RESULTS : To address this issue , purvalanol B ( 95. ) and an N6-methylated , CDK-inactive derivative ( 95M. ) were immobilized on an agarose matrix . Extracts from a diverse collection of cell types and organisms were screened for proteins binding purvalanol B . In addition to validating CDKs as intracellular targets , a variety of unexpected protein kinases were recovered from the 95. matrix . Casein kinase 1 ( CK1 ) was identified as a principal 95. matrix binding protein in Plasmodium falciparum , Leishmania mexicana , Toxoplasma gondii and Trypanosoma cruzi . DB02733 compounds also inhibit the proliferation of these parasites , suggesting that CK1 is a valuable target for further screening with 2,6,9-trisubstituted purine libraries . CONCLUSIONS : That a simple batchwise affinity chromatography approach using two purine derivatives facilitated isolation of a small set of highly purified kinases suggests that this could be a general method for identifying intracellular targets relevant to a particular class of ligands . This method allows a close correlation to be established between the pattern of proteins bound to a small family of related compounds and the pattern of cellular responses to these compounds . [ Low doses of sulphonyluria as a successful replacement for insulin therapy in a patient with neonatal diabetes due to a mutation of Q14654 gene encoding Kir6.2 ] . Neonatal diabetes mellitus is a rare metabolic disorder with an estimated incidence of 1:300.000 to 400.000 newborns , and less than 50 % of the neonates have permanent neonatal diabetes mellitus ( PNDM ) . Recently , activating mutation in the Q14654 gene encoding Kir6.2 subunit of the adenosin triphosphate-sensitive potassium ( K( DB00171 ) ) channel has been described as the most frequent cause of PNDM . Under physiological circumstances K( DB00171 ) channel closure plays a central role in glucose-stimulated insulin secretion from pancreatic beta cells . Sulphonylurea drugs stimulate insulin secretion by binding to and closing K( DB00171 ) channels and thus bypassing beta cell metabolism stimulate the same chain of reactions as glucose . We describe a boy diagnosed with PNDM at the age of 3 months when insulin therapy was started , and at the age of 4.5 years Q14654 gene was sequenced and found that the boy carried a de novo activating R201H mutation . P01308 therapy was successfully switched to low doses of oral glibenclamide . Accordingly , it is important to emphasize that every person diagnosed with diabetes before six months of life , however old they actually are , should be tested for K( DB00171 ) mutations which is offered via the website www.diabetesgenes.org . [ DB06655 ( DB06655 ) : human glucagon-like peptide-1 used in once daily injection for the treatment of type 2 diabetes ] . DB06655 ( DB06655 ) is a peptide produced by DNA recombinant technology , which presents 97 % homology with human glucagon-like peptide-1 ( P0C6A0 ) but is resistant to dipeptidylpeptidase-4 , the enzyme that degrades the natural hormone . It actives the P43220 and exerts an incretin mimetic effect during at least 24 hours after a single subcutaneous injection . Besides a glucose-dependent stimulatory effect of insulin secretion , liraglutide inhibits glucagon secretion and retards gastric emptying . In patients with type 2 diabetes , it reduces glycated haemoglobin by at least 1 % , without inducing hypoglycaemia . It also induces a moderate weight loss and a mild reduction in blood pressure . Gastrointestinal adverse events ( nausea , vomiting ) may occur during the initial phase of treatment , but rarely impose the interruption of the medication and usually diminish with time.Although indicated in combination with other glucose-lowering agents , liraglutide is currently reimbursed in Belgium only if administered in patients with type 2 diabetes not sufficiently controlled with a combination of metformin plus sulfonylurea or metformin plus a thiazolidinedione . DB06655 is presented in prefilled pens and is injected subcutaneously once a day . Treatment will be initiated with 0.6 mg to improve digestive tolerance and the daily dose will be increased to 1.2 mg ( usual dose ) after at least one week , and up to 1.8 mg ( maximal dose ) if necessary . Inhibition of envelope-mediated P01730 +-T-cell depletion by human immunodeficiency virus attachment inhibitors . Human immunodeficiency virus type 1 ( HIV-1 ) envelope ( Env ) binding induces proapoptotic signals in P01730 (+) T cells without a requirement of infection . Defective virus particles , which represent the majority of HIV-1 , usually contain a functional Env and therefore represent a potentially significant cause of such P01730 (+)-T-cell loss . We reasoned that an HIV-1 inhibitor that prohibits Env-host cell interactions could block the destructive effects of defective particles . HIV-1 attachment inhibitors ( AIs ) , which potently inhibit Env- P01730 binding and subsequent downstream effects of Env , display low-nanomolar antiapoptotic potency and prevent P01730 (+)-T-cell depletion from mixed lymphocyte cultures , also with low-nanomolar potency . Specific Env amino acid changes that confer resistance to AI antientry activity eliminate AI antiapoptotic effects . We observed that P01730 (+)-T-cell destruction is specific for P61073 -utilizing HIV-1 strains and that the fusion blocker enfuvirtide inhibits Env-mediated P01730 (+)-T-cell killing but is substantially less potent than AIs . These observations , in conjunction with observed antiapoptotic activities of soluble P01730 and the P61073 blocker DB06809 , suggest that this AI activity functions through a mechanism common to AI antientry activity , e.g. , prevention of Env conformation changes necessary for specific interactions with cellular factors that facilitate viral entry . Our study suggests that AIs , in addition to having potent antientry activity , could contribute to immune system homeostasis in individuals infected with HIV-1 that can engage P61073 , thereby mitigating the increased risk of adverse clinical events observed in such individuals on current antiretroviral regimens . Differential selectivity of insulin secretagogues : mechanisms , clinical implications , and drug interactions . The sulphonylurea receptor ( Q09428 ) subunits of K( DB00171 ) channels are the targets for several classes of therapeutic drugs . Sulphonylureas close K( DB00171 ) channels in pancreatic beta-cells and are used to stimulate insulin release in type 2 diabetes , whereas the K( DB00171 ) channel opener nicorandil acts as an antianginal agent by opening K( DB00171 ) channels in cardiac and vascular smooth muscle . The predominant type of Q09428 varies between tissues : Q09428 in beta-cells , SUR2A in cardiac muscle , and SUR2B in smooth muscle . Sulphonylureas and related drugs exhibit differences in tissue specificity , as the drugs interact to varying degrees with different types of Q09428 . DB01120 and tolbutamide are beta-cell selective and reversible . DB00222 , glibenclamide , and repaglinide , however , inhibit cardiac and smooth muscle K( DB00171 ) channels in addition to those in beta-cells and are only slowly reversible . Similar properties have been observed by recording K( DB00171 ) channel activity in intact cells and in Xenopus oocytes expressing cloned K( DB00171 ) channel subunits . While K( DB00171 ) channels in cardiac and smooth muscle are largely closed under physiological conditions ( but open during ischaemia ) , they are activated by antianginal agents such as nicorandil . Under these conditions , they may be inhibited by sulphonylureas that block SUR2-type K( DB00171 ) channels ( e.g. , glibenclamide ) . Care should , therefore , be taken when choosing a sulphonylurea if potential interactions with cardiac and smooth muscle K( DB00171 ) channels are to be avoided . Differential gene expression in well-regulated and dysregulated pancreatic beta-cell ( MIN6 ) sublines . To identify genes involved in regulated insulin secretion , we have established and characterized two sublines derived from the mouse pancreatic beta-cell line MIN6 , designated B1 and P01024 . They have a similar insulin content , but differ in their secretory properties . B1 responded to glucose in a concentration- and cell confluence-dependent manner , whereas P01024 did not . B1 cells were stimulated by phorbol 12-myristate 13-acetate , leucine , arginine , glibenclamide , isobutylmethylxanthine , and DB00761 , whereas P01024 did not respond ( leucine , arginine , and glibenclamide ) or responded to a lesser extent ( isobutylmethylxanthine , phorbol 12-myristate 13-acetate , and DB00761 ) . Although intracellular Ca(2+) rose in response to glucose in B1 but not P01024 cells , DB00761 increased intracellular Ca(2+) in a similar manner in both sublines . P11166 , P11168 , Kir6.2 , and Q09428 expression was not significantly different between B1 and P01024 cells , whereas P12830 was more abundantly expressed in B1 cells . A more complete list of differentially expressed genes was established by suppression subtractive hybridization and high density ( Affymetrix ) oligonucleotide microarrays . Genes were clustered according to known or putative function . Those involved in metabolism , intracellular signaling , cytoarchitecture , and cell adhesion are of potential interest . These two sublines should be useful for identification of the genes and mechanisms involved in regulated insulin secretion of the pancreatic beta-cell . P10275 inducing bladder cancer progression by promoting an epithelial-mesenchymal transition . The study investigated the role of androgen receptor ( AR ) as a potential target for the treatment of bladder cancer in regulating epithelial-mesenchymal transition or transformation ( EMT ) . Cell proliferation , and migration capacity were determined in bladder cancer T24 cells treated with small interfering RNA directed against AR , and expression levels of P12830 , β-catenin and N- cadherin were assessed using quantitative reverse transcription PCR ( qRT-PCR ) . Tumour cell growth was evaluated in vivo in T24 tumour-bearing nude mice receiving electroporation-assisted administration of anti-AR small interfering RNA . It was found that low AR expression decreased proliferation and migration of bladder cancer cells . In vivo experiments showed that silencing AR expression significantly suppressed AR-positive bladder tumour growth with decreased cell proliferation . Low AR level of T24 bladder cancer cells treated with DB01541 ( DB02901 ) decreased expression of P12830 , β-catenin and P19022 expression , indicating a strong sensitivity to the EMT and In cells with low AR content , TGF-β induced down-regulation of P12830 and β-catenin . It is concluded that suppression of AR expression decreased the production of TGF-β , inhibiting EMT and bladder cancer cell growth in vitro and in vivo , implying that its use might be a potential therapeutic target for the treatment of bladder cancer . Modeling of Q14654 and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 -sensitive potassium ( K+ DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 ) . The drugs and the compounds were docked to the DB00171 -dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME/Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule . [ Relation between insulin secretion and action -- study of genetic determination ] . BACKGROUND : The disposition index represents insulin secretion related to the degree of insulin sensitivity , being constant for given degree of glucose tolerance . The aim of this study is to discern genetic determinants influencing the value of disposition index , e.g. predisposition to glucose intolerance . METHODS AND RESULTS : Two hundred and four non-diabetic subjects with varied glucose tolerance were divided into groups according to the values of disposition index . DB09341 and lipid metabolism , anthropometric parameters and family history of type 2 diabetes mellitus ( DM2 ) were examined . The genotype frequency of candidate genes was compared between the groups of individuals within the lowest ( Q1 ) and the highest ( Q4 ) quartiles of the disposition index values . Those groups were not different concerning age and female to male ratio . Fasting and stimulated parameters of glucose metabolism and lipid profile were worse in group Q1 compared to group Q4 . Group Q1 is characterized with higher number of individuals with metabolic syndrome and family history of DM2 . The examination of candidate genes revealed the differences in genotype frequency of P07550 ( rs1042714 ) , Q07869 ( rs1800206 ) , Q14654 ( rs5219 ) , and Q8IWU4 ( rs13266634 ) between groups Q1 and Q4 . CONCLUSIONS : Low value of disposition index is related to the deterioration of glucose tolerance and other signs of metabolic syndrome . It is associated with genes affecting insulin secretion and genes related to energy metabolism and obesity . G-protein-coupled receptors and asthma endophenotypes : the cysteinyl leukotriene system in perspective . Genetic variation in specific G-protein coupled receptors ( GPCRs ) is associated with a spectrum of respiratory disease predispositions and drug response phenotypes . Although certain GPCR gene variants can be disease-causing through the expression of inactive , overactive , or constitutively active receptor proteins , many more GPCR gene variants confer risk for potentially deleterious endophenotypes . Endophenotypes are traits , such as bronchiole hyperactivity , atopy , and aspirin intolerant asthma , which have a strong genetic component and are risk factors for a variety of more complex outcomes that may include disease states . GPCR genes implicated in asthma endophenotypes include variants of the cysteinyl leukotriene receptors ( Q9Y271 and Q9NS75 ) , and prostaglandin D2 receptors ( Q13258 and Q9Y5Y4 ) , thromboxane A2 receptor ( P21731 ) , beta2-adrenergic receptor ( P07550 ) , chemokine receptor 5 ( P51681 ) , and the G protein-coupled receptor associated with asthma ( Q6W5P4 ) . This review of the contribution of variability in these genes places the contribution of the cysteinyl leukotriene system to respiratory endophenotypes in perspective . The genetic variant(s) of receptors that are associated with endophenotypes are discussed in the context of the extent to which they contribute to a disease phenotype or altered drug efficacy . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . Regulation of P30518 transcription by hypertonicity and P37288 - P30518 signal interaction . DB00067 ( AVP ) and hypertonicity in the renal medulla play a major role in the urine concentration mechanism . Previously , we showed that rat vasopressin V2 receptor ( rV2R ) promoter activity was increased by vasopressin P30518 stimulation and decreased by vasopressin V1a receptor ( P37288 ) stimulation in a LLC- P30613 cell line stably expressing rat P37288 ( LLC- P30613 /rV1aR ) . In the present study , we investigated the effects of hypertonicity on the rV2R promoter activity and on the suppression of rV2R promoter activity by P37288 stimulation in LLC- P30613 /rV1aR cells . rV2R promoter activity was increased in NaCl- or mannitol-induced hypertonicity . The hypertonicity-responsive site in the rV2R promoter region was limited to 10 bp , including the Sp1 motif . The increase of P30518 promoter activity by hypertonicity was significantly inhibited by a JNK inhibitor ( SP600125 ) and PKA inhibitor ( H89 ) . In contrast , rV2R promoter activity was remarkably suppressed by P37288 stimulation in the hypertonic condition rather than in the isotonic condition . The AVP-stimulated intracellular Ca2+ concentration was increased in the hypertonic condition , suggesting the functional activation of P37288 by hypertonicity . In conclusion , 1 ) P30518 promoter activity is increased by hypertonicity via the JNK and PKA pathways , 2 ) suppression of P30518 expression by the P37288 -Ca2+ pathway is enhanced by hypertonicity , and 3 ) hypertonicity enhances the P37288 -Ca2+ pathway . The counteractivity of P30518 and P37288 could be required to maintain minimum urine volume in the dehydrated state . A circuitry and biochemical basis for tuberous sclerosis symptoms : from epilepsy to neurocognitive deficits . Tuberous sclerosis complex ( TSC ) is an autosomal dominant monogenetic disorder that is characterized by the formation of benign tumors in several organs as well as brain malformations and neuronal defects . TSC is caused by inactivating mutations in one of two genes , Q92574 and P49815 , resulting in increased activity of the mammalian Target of DB00877 ( P42345 ) . Here , we explore the cytoarchitectural and functional CNS aberrations that may account for the neurological presentations of TSC , notably seizures , hydrocephalus , and cognitive and psychological impairments . In particular , recent mouse models of brain lesions are presented with an emphasis on using electroporation to allow the generation of discrete lesions resulting from loss of heterozygosity during perinatal development . Cortical lesions are thought to contribute to epileptogenesis and worsening of cognitive defects . However , it has recently been suggested that being born with a mutant allele without loss of heterozygosity and associated cortical lesions is sufficient to generate cognitive and neuropsychiatric problems . We will thus discuss the function of P42345 hyperactivity on neuronal circuit formation and the potential consequences of being born heterozygous on neuronal function and the biochemistry of synaptic plasticity , the cellular substrate of learning and memory . Ultimately , a major goal of TSC research is to identify the cellular and molecular mechanisms downstream of P42345 underlying the neurological manifestations observed in TSC patients and identify novel therapeutic targets to prevent the formation of brain lesions and restore neuronal function . The Drosophila insulin receptor activates multiple signaling pathways but requires insulin receptor substrate proteins for DNA synthesis . The Drosophila insulin receptor ( P30518 ) contains a 368-amino-acid COOH-terminal extension that contains several tyrosine phosphorylation sites in YXXM motifs . This extension is absent from the human insulin receptor but resembles a region in insulin receptor substrate ( P41252 ) proteins which binds to the phosphatidylinositol ( PI ) 3-kinase and mediates mitogenesis . The function of a chimeric P30518 containing the human insulin receptor binding domain ( hDIR ) was investigated in 32D cells , which contain few insulin receptors and no P41252 proteins . P01308 stimulated tyrosine autophosphorylation of the human insulin receptor and hDIR , and both receptors mediated tyrosine phosphorylation of Shc and activated mitogen-activated protein kinase . P35568 was required by the human insulin receptor to activate PI 3-kinase and p70s6k , whereas hDIR associated with PI 3-kinase and activated p70s6k without P35568 . However , both receptors required P35568 to mediate insulin-stimulated mitogenesis . These data demonstrate that the P30518 possesses additional signaling capabilities compared with its mammalian counterpart but still requires P35568 for the complete insulin response in mammalian cells . Regulation of glucagon-like peptide-1 receptor and calcium-sensing receptor signaling by L-histidine . Receptor-specific agonists of the extracellular calcium-sensing receptor ( P41180 ) potentiate glucose-induced insulin secretion , an effect similar to that of glucagon-like peptide-1 ( P0C6A0 ) . We have sequenced the full open reading frame of the P41180 from rat insulinoma ( P01308 -1 ) cells and find that the predicted amino acid sequence of the receptor is identical with that of the receptor from the parathyroid gland . This receptor couples to both Gq/11 and Gi/o , and this dual coupling may partly explain the varying effects of nonspecific agonists on secretion reported previously . L- DB00117 ( L- DB00117 ) increases the sensitivity of the P41180 to extracellular Ca2+ and potentiates glucose-dependent insulin secretion from P01308 -1 cells . This potentiation is partially inhibited at low extracellular [ Ca2+ ] where the P41180 is ineffective . Coexpression of the P41180 and P43220 ( P43220 ) produces a pertussis toxin-sensitive inhibition of P0C6A0 -induced DB02527 production in response to elevated extracellular [ Ca2+ ] . However , l- DB00117 potentiates DB02527 response element reporter activity in P01308 -1 cells and in human embryonic kidney-293 cells expressing either the P43220 alone or the P41180 and P43220 . P01308 -1 cells express the RNA for the P41180 at a lower level than that for the P43220 . This difference in expression level of the receptors may explain the potentiation of insulin secretion by L- DB00117 despite coupling of the P41180 to Gi/o . In conclusion , L- DB00117 can potentiate both P43220 - and P41180 -activated signaling pathways , and these effects may play a role in the potentiation of glucose-induced insulin secretion in response to meals containing protein in addition to carbohydrates and fat . Astaxanthin improves the proliferative capacity as well as the osteogenic and adipogenic differentiation potential in neural stem cells . In the present study , the effect of astaxanthin on improvement of the proliferative capacity as well as the osteogenic and adipogenic differentiation potential in neural stem cells ( NSCs ) was evaluated . Treatment of astaxanthin-induced actives cell growth in a dose-dependent and time-dependent manner . Results from a clonogenic assay clearly indicated that astaxanthin can actively stimulate proliferation of NSCs . Astaxanthin-induced improvement in the proliferative capacity of NSCs resulted in overexpression of several proliferation-related proteins . Astaxanthin-induced activation of PI3K and its downstream mediators , p-MEK , p- P29323 , and p-Stat3 in NSCs resulted in subsequent induction of expression of proliferation-related transcription factors ( Rex1 , P06493 , and P24941 ) and stemness genes ( Q01860 , P48431 , Nanog , and O43474 ) . Astaxanthin also improved the osteogenic and adipogenic differentiation potential of NSCs . Astaxanthin-treated NSCs showed prominent calcium deposits and fat formation . These results were consistent with overexpression of osteogenesis-related genes ( osteonectin , RXR , and osteopontin ) and adipogenesis-related genes ( AP and P37231 ) after astaxanthin treatment . These findings clearly demonstrated that astaxanthin acts synergistically on the regulatory circuitry that controls proliferation and differentiation of NSCs . Developmental programming : effect of prenatal steroid excess on intraovarian components of insulin signaling pathway and related proteins in sheep . Prenatal testosterone ( T ) excess increases ovarian follicular recruitment , follicular persistence , insulin resistance , and compensatory hyperinsulinemia . Considering the importance of insulin in ovarian physiology , in this study , using prenatal T- and dihydrotestosterone ( DB02901 , a nonaromatizable androgen ) -treated female sheep , we tested the hypothesis that prenatal androgen excess alters the intraovarian insulin signaling cascade and metabolic mediators that have an impact on insulin signaling . Changes in ovarian insulin receptor ( INSRB ) , insulin receptor substrate 1 ( P35568 ) , mammalian target of rapamycin ( P42345 ) , phosphatidylinositol 3-kinase ( PIK3 ) , peroxisome proliferator-activated receptor-gamma ( P37231 ) , and adiponectin proteins were determined at fetal ( Days 90 and 140 ) , postpubertal ( 10 mo ) , and adult ( 21 mo ) ages by immunohistochemistry . Results indicated that these proteins were expressed in granulosa , theca , and stromal compartments , with INSRB , P35568 , P37231 , and adiponectin increasing in parallel with advanced follicular differentiation . Importantly , prenatal T excess induced age-specific changes in P37231 and adiponectin expression , with increased P37231 expression evident during fetal life and decreased antral follicular adiponectin expression during adult life . Comparison of developmental changes in prenatal T and DB02901 -treated females found that the effects on P37231 were programmed by androgenic actions of T , whereas the effects on adiponectin were likely by its estrogenic action . These results suggest a role for P37231 in the programming of ovarian disruptions by prenatal T excess , including a decrease in antral follicular adiponectin expression and a contributory role for adiponectin in follicular persistence and ovulatory failure . Q9Y271 is involved in N-methyl-D-aspartate-mediated neuronal injury in mice . AIM : To determine whether cysteinyl leukotriene receptor 1 ( CysLT1 receptor ) is involved in N-methyl-D-aspartate ( DB01221 ) -induced excitotoxic injury in the mouse brain . METHODS : Brain injury was induced by DB01221 microinjection ( 50-150 nmol in 0.5 microL ) into the cerebral cortex . The changes in CysLT1 receptor expression 24 h after DB01221 injection and the effects of a CysLT1 receptor antagonist , pranlukast ( 0.01 and 0.1 mg/kg ) , an DB01221 receptor antagonist , ketamine ( 30 mg/kg ) , and an antioxidant , edaravone ( 9 mg/kg ) were observed . RESULTS : In the DB01221 -injured brain , the CysLT1 receptor mRNA , and protein expression were upregulated , and the receptor was mainly localized in the neurons and not in the astrocytes . DB01411 , ketamine and edaravone decreased DB01221 -induced injury ; pranlukast ( 0.1 mg/kg ) and ketamine inhibited the upregulated expression of the CysLT1 receptor . CONCLUSION : CysLT1 receptor expression in neurons is upregulated after DB01221 injection , and DB01221 -induced responses are inhibited by CysLT1 receptor antagonists , indicating that the increased CysLT1 receptor is involved in DB01221 excitotoxicity . A Short-activating RNA Oligonucleotide Targeting the Islet β-cell Transcriptional Factor MafA in P28906 (+) Cells . Upon functional loss of insulin producing islet β-cells , some patients with diabetes become dependent on life-long insulin supplementation therapy . Bioengineering surrogate insulin producing cells is an alternative replacement strategy . We have developed a novel approach using short-activating RNA oligonucleotides to differentiate adult human P28906 (+) cells into insulin-secreting cells . By transfecting RNA to increase transcript levels of the master regulator of insulin biosynthesis , v-maf musculoaponeurotic fibrosarcoma oncogene homolog A ( MafA ) , several pancreatic endodermal genes were upregulated during the differentiation procedure . These included Pancreatic and duodenal homeobox gene-1 ( PDX1 ) , Neurogenin 3 , Q13562 , and NK6 homeobox 1 ( NKx6-1 ) . Differentiated P28906 (+) cells also expressed glucokinase , glucagon-like peptide 1 receptor ( P43220 ) , sulfonylurea receptor-1 ( Q09428 ) and phogrin-all essential for glucose sensitivity and insulin secretion . The differentiated cells appropriately processed C-peptide and insulin in response to increasing glucose stimulation as shown by enzyme-linked immunosorbent assay ( ELISA ) , fluorescence-activated cell sorting analysis , western blotting , and immunofluorescence staining . We provide a new approach using short-activating RNA in developing insulin producing surrogate cells for treating diabetes.Molecular Therapy - Nucleic Acids ( 2013 ) 2 , e97 ; doi:10.1038/mtna.2013.23 ; advance online publication 4 June 2013 . Characterization of the effect of chronic administration of a calcium-sensing receptor antagonist , DB05255 , on renal calcium excretion and serum calcium in postmenopausal women . Ronacaleret is an orally-active calcium-sensing receptor ( P41180 ) antagonist that has the potential for therapeutic utility in the stimulation of PTH release , notably as a bone anabolic agent comparable to recombinant human PTH(1-34) ( DB05829 (1-34) ) . A recent study has shown that , despite the ability to increase circulating PTH levels in postmenopausal women in a dose-dependent manner , minimal effects of DB05255 on bone mineral density have been observed . Therefore , the purpose of this study was to characterize the PTH profile as well as calcium metabolism parameters as a marker of PTH biological activity following the administration of DB05255 or DB05829 (1-34) . Administration of DB05255 led to lower peak levels of PTH than were observed with DB05829 (1-34) , however , greater total PTH exposure was observed . Further , chronic administration of either agent was associated with increases in urinary calcium excretion and serum calcium levels , with the magnitude of the changes following DB05255 significantly greater than that for DB05829 (1-34) . The greater magnitude of effects observed with DB05255 is likely due to the greater total PTH exposure , and is potentially reflective of a state comparable to mild hyperparathyroidism . It is not clear whether the administration of all calcilytics would lead to a similar result , or is due to characteristics specific to DB05255 . Monoclonal antibodies targeting P01584 reduce biomarkers of atherosclerosis in vitro and inhibit atherosclerotic plaque formation in P02649 -deficient mice . OBJECTIVE : Atherosclerosis is a condition that is increasingly contributing to worldwide mortality through complications such as stroke and myocardial infarction . IL-1β plays multiple direct , local roles in the formation and stability of the atheroma by eliciting the production of additional cytokines and proteolytic enzymes from macrophages , endothelial cells ( EC ) and smooth muscle cells ( SMC ) . We therefore tested whether an anti-IL-1β antibody , DB06062 , might inhibit the secretion of pro-atherogenic cytokines from macrophages in vitro and affect a positive outcome in the P02649 -deficient mouse ( ApoE(-/-) ) model of atherosclerosis in vivo . METHODS AND RESULTS : In an in vitro co-culture model , DB06062 inhibited macrophage-induced secretion of key atherogenic cytokines from EC and SMC , including P05231 , P10145 , P13500 and TNFα . The release of degradative enzymes , such as the matrix metalloproteinases P08254 and P14780 , was also decreased by DB06062 . In addition , DB06062 inhibited the secretion of P13232 from EC and P05112 from SMC , cytokines not previously reported to be driven by IL-1β in this context . In vivo , XMA052 MG1K , a chimeric murine version of DB06062 , inhibited the formation of atherosclerotic lesions in the ApoE(-/-) model at all three doses tested . This effect was comparable to that reported for complete genetic ablation of IL-1β or IL-1R1 on an ApoE(-/-) background and was associated with decreases in plasma non-HDL/HDL cholesterol ratio and plaque lipid content and macrophage infiltration . CONCLUSIONS : These results demonstrate for the first time that an antibody targeting IL-1β can inhibit the progression of atherosclerosis in vivo , highlighting the importance of this key cytokine in cardiovascular disease . The efficacy of combination antiretroviral therapy in HIV type 1-infected patients treated in Curaçao compared with Antillean , Surinam , and Dutch HIV type 1-infected patients treated in The Netherlands . We compared the efficacy of combination antiretroviral therapy ( cART ) of Antillean HIV-1-infected patients treated on the Caribbean island of Curaçao ( CUR-AN ) with Antillean ( NL-AN ) , Surinam ( NL- Q09428 ) , and Dutch ( NL-NL ) patients treated in The Netherlands . In total 2118 therapy-naive patients who started cART between January 2005 and August 2008 were included in the comparison . The CUR-AN patients initiated cART at a median P01730 cell count of 141 cells/mm(3) and 63 % had counts below 200 cells/mm(3) . Within 12 months of the start of cART 76 % of the CUR-AN patients achieved viral suppression , defined as HIV-1 RNA plasma levels below 80 copies/ml . The percentage achieving viral suppression was higher in patients treated in The Netherlands ( NL-AN = 87 % , NL- Q09428 = 93 % , and NL-NL = 96 % ) . Lost to follow-up after 30 months of cART was 10 % among CUR-AN patients and was higher than observed among patients treated in The Netherlands ( NL-AN = 8 % , NL- Q09428 = 3 % , and NL-NL = 2 % ) . A similar pattern was found for progression to AIDS and death ( 10 % of CUR-AN vs. 5 % , 6 % , and 7 % of NL-AN , NL- Q09428 , and NL-NL patients , respectively ) . Late start of cART and limited viral suppression after the start of cART determine the higher rate of disease progression to AIDS and death among Antillean patients treated in Curaçao . The high percentage of lost to follow-up may result in an underestimation of AIDS and AIDS-related death among HIV-1-infected Antilleans treated in Curaçao . Screening of 134 single nucleotide polymorphisms ( SNPs ) previously associated with type 2 diabetes replicates association with 12 SNPs in nine genes . More than 120 published reports have described associations between single nucleotide polymorphisms ( SNPs ) and type 2 diabetes . However , multiple studies of the same variant have often been discordant . From a literature search , we identified previously reported type 2 diabetes-associated SNPs . We initially genotyped 134 SNPs on 786 index case subjects from type 2 diabetes families and 617 control subjects with normal glucose tolerance from Finland and excluded from analysis 20 SNPs in strong linkage disequilibrium ( r(2) > 0.8 ) with another typed SNP . Of the 114 SNPs examined , we followed up the 20 most significant SNPs ( P < 0.10 ) on an additional 384 case subjects and 366 control subjects from a population-based study in Finland . In the combined data , we replicated association ( P < 0.05 ) for 12 SNPs : P37231 Pro12Ala and His447 , Q14654 Glu23Lys and rs5210 , P01375 -857 , P11168 Ile110Thr , P20823 /TCF1 rs2701175 and GE117881_360 , P35558 -232 , Q13562 Thr45Ala , P05231 -598 , and P22413 Lys121Gln . The replication of 12 SNPs of 114 tested was significantly greater than expected by chance under the null hypothesis of no association ( P = 0.012 ) . We observed that SNPs from genes that had three or more previous reports of association were significantly more likely to be replicated in our sample ( P = 0.03 ) , although we also replicated 4 of 58 SNPs from genes that had only one previous report of association .

MH_train_1344

MH_train_1344

MH_train_1344

interacts_with DB00559?

multiple_choice

Endothelial progenitor cells in relation to endothelin-1 and endothelin receptor blockade : a randomized , controlled trial . AIMS : Endothelial progenitor cells ( EPC ) represent an endogenous repair mechanism involving rendothelialization and neoangiogenesis . Patients with both diabetes and vascular disease have low numbers of circulating EPC . The endothelium-derived peptide , endothelin-1 ( ET-1 ) , is increased in patients with type 2 diabetes and vascular complications and has been suggested to contribute to endothelial dysfunction . Therefore , we investigated the relation between EPC and plasma ET-1 and the effect of dual ET-1 receptor antagonist treatment . METHODS : In this double blind study patients with type 2 diabetes mellitus and microalbuminuria were randomized to treatment with the dual P25101 /ETB receptor antagonist DB00559 treatment ( 125mg bid ; n=17 ) or placebo ( n=19 ) for four weeks . Different EPC subpopulations were enumerated by flow cytometry using triple staining ( P28906 , CD133 , P35968 ) at baseline at the end of treatment . Viability was assessed by 7AAD and Annexin-V-staining . RESULTS : Baseline ET-1 levels correlated significantly with P02741 levels . Patients with ET-1 levels above the median value had higher levels of P28906 (+)CD133(+) and P28906 (+) P35968 (+) EPC . There was no difference in P28906 (+) and P28906 (+)CD133(+) P35968 (+) cells , markers of EPC apoptosis or circulating markers of endothelial damage between patients with ET-1 levels below or above the median . Four week treatment with DB00559 did not change EPC levels . CONCLUSION : Among patients with type 2 diabetes and vascular disease , high plasma levels of ET-1 are associated with higher number of EPC . The recruitment of EPC does not seem to be regulated via ET-1 receptor activation since treatment with a dual ET-1 receptor blocker did not affect circulating EPC numbers . Population pharmacokinetics of ethionamide in patients with tuberculosis . SETTING : Three US referral hospitals . OBJECTIVE : Determine the population pharmacokinetic ( PK ) parameters of ethionamide ( P25101 ) following multiple oral doses . DESIGN : Fifty-five patients with tuberculosis ( TB ) participated . Patients received multiple oral doses of P25101 as part of their treatment . They also received other anti-tuberculosis medications based upon in vitro susceptibility data . Serum samples were collected over 12 h post-dose , and concentrations were determined using a validated high-performance liquid chromatography ( HPLC ) assay . Concentration-time data were analyzed using population methods . RESULTS : P25101 areas under the concentration-versus-time curve ( AUCs ) increased linearly with increasing oral doses from 250 to 1000 mg . Compared to the population pattern , delayed absorption was seen at least once in 15 % of patients . P25101 PK parameter estimates were independent of age , weight , height , gender , and creatinine clearance . TB patients appeared to have larger volumes of distribution ( 3.22 l/kg ) and clearance values ( 1.88 l/h/kg ) compared to previously studied healthy volunteers . This resulted in lower AUC values ( 3.95 mcg h/ml ) in the TB patients . P25101 displayed a short elimination half-life ( 1.94 h ) . The effect of different dosing strategies on calculated pharmacodynamic parameters was explored . Simulated doses of 250 mg P55957 to TID failed to achieve serum concentrations above the MIC . CONCLUSION : P25101 PK parameters differed between TB patients and healthy volunteers , possibly due to differences in the completeness of absorption . Doses of at least 500 mg appear to be required to achieve serum concentrations above the typical P25101 MIC . Additional research is needed to determine the optimal dosing of P25101 . Enhanced expression of pro-inflammatory mediators and liver Q9UBH6 -regulated lipogenic genes in non-alcoholic fatty liver disease and hepatitis C . NAFLD ( non-alcoholic fatty liver disease ) is one of the most frequent chronic liver diseases worldwide . The metabolic factors associated with NAFLD are also determinants of liver disease progression in chronic HCV ( hepatitis C virus ) infection . It has been reported that , besides inducing hepatic fatty acid biosynthesis , LXR ( liver X receptor ) regulates a set of inflammatory genes . We aimed to evaluate the hepatic expression of LXRα and its lipogenic and inflammatory targets in 43 patients with NAFLD , 44 with chronic HCV infection and in 22 with histologically normal liver . Real-time PCR and Western blot analysis were used to determine hepatic expression levels of LXRα and related lipogenic and inflammatory mediators in the study population . We found that the LXRα gene and its lipogenic targets Q07869 -γ ( peroxisome-proliferator-activated receptor-γ ) , SREBP ( sterol-regulatory-element-binding protein ) -1c , Q12772 and FAS ( fatty acid synthase ) were overexpressed in the liver of NAFLD and HCV patients who had steatosis . Moreover , up-regulation of inflammatory genes , such as P01375 ( tumour necrosis factor ) -α , IL (interleukin)-6 , P10451 ( osteopontin ) , P35228 ( inducible NO synthase ) , P36551 (cyclo-oxygenase)-2 and Q9NSE2 ( suppressors of cytokine signalling ) -3 , was observed in NAFLD and HCV patients . Interestingly , P01375 -α , P05231 and osteopontin gene expression was lower in patients with steatohepatitis than in those with steatosis . In conclusion , hepatic expression of LXRα and its related lipogenic and inflammatory genes is abnormally increased in NAFLD and HCV patients with steatosis , suggesting a potential role of LXRα in the pathogenesis of hepatic steatosis in these chronic liver diseases . Dual endothelin receptor antagonism prevents remodeling of resistance arteries in diabetes . Vascular remodeling , characterized by extracellular matrix deposition and increased media-to-lumen ( M/L ) ratio , contributes to the development of microvascular complications in diabetes . We have previously shown in type 2 diabetic Goto-Kakizaki ( GK ) rats that selective P25101 receptor blockade prevents medial thickening of mesenteric arteries via regulation of matrix metalloproteases ( MMP ) , whereas selective ETB receptor blockade augments this thickening . The goal of this study was to determine the effect of combined P25101 and ETB receptor blockade on resistance vessel remodeling . Vessel structure , MMP activity , and extracellular matrix proteins were assessed in control Wistar and diabetic GK rats treated with vehicle or DB00559 ( 100 mg/kg per day ) for 4 weeks ( n = 7-9 per group ) . DB00559 completely prevented the increase in M/L ratio and P08253 activity in diabetes but paradoxically increased M/L ratio and MMP activation in control animals . Collagenase ( P45452 ) activity and protein levels were significantly decreased in diabetes . Accordingly , collagen deposition was augmented in GK rats . Dual ET receptor antagonism improved enzyme activity and normalized P45452 levels in diabetic animals but blunted P45452 activity in control animals . In summary , current findings suggest that diabetes-mediated remodeling of resistance arteries is prevented by dual blockade of P25101 and ETB receptors and that the relative role of ET receptors in the regulation of vascular structure differs in the control and disease states . Key residues at the riboflavin kinase catalytic site of the bifunctional riboflavin kinase/ Q8NFF5 from Corynebacterium ammoniagenes . Many known prokaryotic organisms depend on a single bifunctional enzyme , encoded by the RibC of RibF gene and named DB03147 synthetase ( FADS ) , to convert DB00140 ( RF ) , first into Q68DA7 and then into DB03147 . The reaction occurs through the sequential action of two activities present on a single polypeptide chain where the N-terminus is responsible for the DB00171 : Q8NFF5 ( FMNAT ) activity and the C-terminus for the DB00171 : riboflavin kinase ( Q969G6 ) activity . Sequence and structural analysis suggest that T208 , N210 and E268 at the C-terminus Q969G6 module of Corynebacterium ammoniagenes FADS ( CaFADS ) might be key during RF phosphorylation . The effect of site-directed mutagenesis on the Q969G6 activity , as well as on substrates and products binding , indicates that T208 and N210 provide the Q969G6 active-site geometry for binding and catalysis , while E268 might be involved in the catalytic step as catalytic base . These data additionally suggest concerted conformational changes at the Q969G6 module of CaFADS during its activity . Mutations at the Q969G6 site also modulate the binding parameters at the FMNAT active site of CaFADS , altering the catalytic efficiency in the transformation of Q68DA7 into DB03147 . This observation supports the hypothesis that the hexameric assembly previously revealed by the crystal structure of CaFADS might play a functional role during catalysis . Immune Checkpoint Blockade in Cancer Therapy . Immunologic checkpoint blockade with antibodies that target cytotoxic T lymphocyte-associated antigen 4 ( P16410 ) and the programmed cell death protein 1 pathway ( P18621 / Q9NZQ7 ) have demonstrated promise in a variety of malignancies . DB06186 ( P16410 ) and pembrolizumab ( P18621 ) are approved by the US Food and Drug Administration for the treatment of advanced melanoma , and additional regulatory approvals are expected across the oncologic spectrum for a variety of other agents that target these pathways . Treatment with both P16410 and P18621 / Q9NZQ7 blockade is associated with a unique pattern of adverse events called immune-related adverse events , and occasionally , unusual kinetics of tumor response are seen . Combination approaches involving P16410 and P18621 / Q9NZQ7 blockade are being investigated to determine whether they enhance the efficacy of either approach alone . Principles learned during the development of P16410 and P18621 / Q9NZQ7 approaches will likely be used as new immunologic checkpoint blocking antibodies begin clinical investigation . Distinct functions of activated protein C differentially attenuate acute kidney injury . Administration of activated protein C ( P25054 ) protects from renal dysfunction , but the underlying mechanism is unknown . P25054 exerts both antithrombotic and cytoprotective properties , the latter via modulation of protease-activated receptor-1 ( P25116 ) signaling . We generated P25054 variants to study the relative importance of the two functions of P25054 in a model of LPS-induced renal microvascular dysfunction . Compared with wild-type P25054 , the K193E variant exhibited impaired anticoagulant activity but retained the ability to mediate P25116 -dependent signaling . In contrast , the L8W variant retained anticoagulant activity but lost its ability to modulate P25116 . By administering wild-type P25054 or these mutants in a rat model of LPS-induced injury , we found that the P25116 agonism , but not the anticoagulant function of P25054 , reversed LPS-induced systemic hypotension . In contrast , both functions of P25054 played a role in reversing LPS-induced decreases in renal blood flow and volume , although the effects on P25116 -dependent signaling were more potent . Regarding potential mechanisms for these findings , P25054 -mediated P25116 agonism suppressed LPS-induced increases in the vasoactive peptide adrenomedullin and infiltration of P35228 -positive leukocytes into renal tissue . However , the anticoagulant function of P25054 was responsible for suppressing LPS-induced stimulation of the proinflammatory mediators P12821 -1 , P05231 , and Q14116 , perhaps accounting for its ability to modulate renal hemodynamics . Both variants reduced active caspase-3 and abrogated LPS-induced renal dysfunction and pathology . We conclude that although P25116 agonism is solely responsible for P25054 -mediated improvement in systemic hemodynamics , both functions of P25054 play distinct roles in attenuating the response to injury in the kidney . Increase in hepatic expression of Q12772 by gemfibrozil administration to rats . It is well known that gemfibrozil increases the biliary output of cholesterol and phospholipids , but we have little knowledge about the impact these changes have on liver cholesterol and phospholipid biosynthetic pathways . In the present study , no changes were detected in liver lipids and P53007 :phosphocholine cytidylyltransferase after gemfibrozil administration to rats . On the contrary , 3-hydroxy-3-methylglutaryl- DB01992 reductase mRNA ( 9.9-fold ) and Rd activity ( 16.7-fold ) and phosphatidate phosphohydrolase activity ( 1.7-fold ) increased , while plasma apo B-cholesterol ( 40 % ) and triglyceride ( 43 % ) levels decreased . As a part of a compensatory homeostatic response , we report for the first time that gemfibrozil administration to rats increased the hepatic sterol regulatory element binding protein-2 ( Q12772 ) mRNA ( 2.9-fold ) and mature protein ( 2.2-fold ) levels . An early increase in the transcriptional activity of Q12772 elicited by gemfibrozil administration might be responsible for the observed changes in P04035 , phosphatidate phosphohydrolase , and Q12772 expression . Role of Q14116 in overt pain-like behaviour in mice . There are evidences that targeting Q14116 might be beneficial to inhibit inflammatory symptoms , including hypernociception ( decrease in nociceptive threshold ) . The mechanism of Q14116 mechanical hypernociception depends on endothelin in rats and mice . However , the role of Q14116 in overt pain-like behaviour remains undetermined . Therefore , we addressed the role of Q14116 in writhing response induced by intraperitoneal ( i.p. ) injection of phenyl-p-benzoquinone ( PBQ ) and acetic acid in mice . Firstly , it was detected that PBQ and acetic acid i.p. injection induced a dose-dependent number of writhes in Balb/c mice . Subsequently , it was observed that the PBQ - but not the acetic acid-induced writhes were diminished in Q14116 deficient ( ( -/- ) ) mice . Therefore , considering that P01579 , endothelin and prostanoids mediate Q14116 -induced mechanical hypernociception , we also investigated the role of these mediators in the same model of writhing response in which Q14116 participates . It was noticed that PBQ-induced writhes were diminished in P01579 (-/-) mice and by the treatment with DB00559 ( mixed endothelin P25101 /ETB receptor antagonist ) , BQ 123 ( cyclo[DTrp-DAsp-Pro-DVal- DB00149 ] , selective endothelin P25101 receptor antagonist ) , BQ 788 ( N-cys-2,6 dimethylpiperidinocarbonyl-l-methylleucyl-d-1-methoxycarboyl-d-norleucine , selective endothelin ETB receptor antagonist ) or indomethacin ( cycloxigenase inhibitor ) . Thus , Q14116 , P01579 , endothelin acting on endothelin P25101 and ETB receptors , and prostanoids mediate PBQ-induced writhing response in mice . To conclude , these results further advance the understanding of the physiopathology of overt pain-like behaviour , and suggest for the first time a role for Q14116 in writhing response in mice . NT-702 ( parogrelil hydrochloride , DB05505 ) , a novel and potent phosphodiesterase inhibitor , improves reduced walking distance and lowered hindlimb plantar surface temperature in a rat experimental intermittent claudication model . NT-702 ( parogrelil hydrochloride , DB05505 ) , 4-bromo-6-[3-(4-chlorophenyl)propoxy]-5-[(pyridin-3-ylmethyl)amino]pyridazin-3(2H)-one hydrochloride , a novel phosphodiesterase ( PDE ) inhibitor synthesized as a potent vasodilatory and antiplatelet agent , is being developed for the treatment of intermittent claudication ( IC ) in patients with peripheral arterial disease . We assessed the efficacy of NT-702 in an experimental IC model as compared with cilostazol and additionally investigated the pharmacological property in vitro and ex vivo . NT-702 selectively inhibited PDE3 ( IC(50)=0.179 and 0.260 nM for Q14432 and 3B ) more potently than cilostazol ( IC(50)=231 and 237 nM for Q14432 and 3B ) among recombinant human PDE1 to PDE6 . NT-702 inhibited in vitro human platelet aggregation induced by various agonists ( IC(50)=11 to 67 nM ) and phenylephrine-induced rat aortic contraction ( IC(50)=24 nM ) . Corresponding results for cilostazol were 4.1 to 17 microM and 1.0 microM , respectively . NT-702 ( 3 mg/kg or more ) significantly inhibited ex vivo rat platelet aggregation after a single oral dose . For cilostazol , 300 mg/kg was effective . In a rat femoral artery ligation model , NT-702 at 5 and 10 mg/kg repeated oral doses twice a day ( P55957 ) for 13 days significantly improved the reduced walking distance while the lowered plantar surface temperature was improved at 2.5 mg/kg and more . DB01166 also improved the walking distance and surface temperature at 300 mg/kg P55957 but significant difference was only observed for surface temperature on day 8 . These results suggest that NT-702 can be expected to have therapeutic advantage for IC . Cellular retinol-binding proteins are determinants of retinol uptake and metabolism in stably transfected Caco-2 cells . The mammalian small intestine contains two related cellular retinol-binding proteins , P09455 and P09455 II , which are thought to have distinct functions . The human intestinal cell line , Caco-2 , was used as a model system for testing the hypothesis that intracellular levels of these proteins directly modulate the absorption and subsequent metabolism of retinol in enterocytes . Immunoblot and Northern blot hybridization demonstrated that Caco-2 cells express P09455 II and cellular retinoic acid-binding protein I in increasing amounts as the cells become more differentiated but do not express detectable quantities of P09455 . Stably transfected cloned Caco-2 cell lines that over-express P09455 II or coexpress P09455 and P09455 II and a control cell line that contains the expression vector without an insert were established . DB00162 uptake and retinyl ester synthesis were increased up to 2-fold by coexpression of P09455 or over-expression of P09455 II . No significant differences were detected in the pattern of retinyl esters synthesized from exogenous [3H]retinol . This suggests that the fatty acid pools utilized for retinol esterification were the same despite differences in the P09455 and P09455 II phenotype of the cell lines . There were no differences between apical and basolateral [3H]retinol uptake or metabolism for filter grown transfected or wild type cell monolayers . Thus , neither P09455 II nor P09455 appear to preferentially interact with luminal- or plasma-derived retinol . Notably , in a cell line which over-expressed P09455 , endogenous P09455 II was reduced by 5-10-fold compared with the wild type and control cell lines . These studies indicate that P09455 and P09455 II levels are determinants of intracellular retinol accumulation and esterification , and they suggest that P09455 -bound retinol or a metabolite can regulate the expression of P09455 II in the mammalian intestine . Blood flow alterations in TNBS-induced colitis : role of endothelin receptors . OBJECTIVES : The aim of the present study was to investigate the time dependent changes in hemodynamic parameters and to assess the role of endothelin ( ET ) receptors in trinitrobenzene sulfonic acid ( TNBS ) induced colitis . MATERIALS : Inferior mesenteric artery ( IMA ) hemodynamics , myeloperoxidase activity ( P05164 ) and damage scores were measured immediately or 1 , 3 , 5 and 14 days after colitis . TREATMENTS : Another group of rats received a nonselective ET receptor antagonist DB00559 ( 30 mg/kg/day ) , P25101 receptor antagonist BQ485 ( 60 microg/rat/day ) or P24530 receptor antagonist BQ788 ( 60 microg/rat/day ) prior to and on the 1st , 2nd and 3rd days after TNBS administration . RESULTS : IMA flow significantly increased at 90 min followed by a substantial decrease through days 1-5 . Tissue P05164 activity and macroscopic damage score increased on 1st day after the induction of colitis and remained elevated 3 , 5 and 14 days following colitis . Treatment with DB00559 or P25101 receptor antagonist largely prevented the colitis-induced reduction in blood flow and tissue injury whereas P24530 receptor antagonist did not attenuate tissue injury or reductions in blood flow . CONCLUSIONS : Our results demonstrate that time-dependent abnormalities occur in IMA hemodynamics following TNBS administration . Our findings also indicate that P25101 receptors but not P24530 receptors play an important role in the colonic inflammation following TNBS administration . DB05007 -- a multitargeted tyrosine kinase inhibitor : results of a phase II study in subjects with non-small cell lung cancer who have progressed after responding to treatment with either gefitinib or erlotinib . INTRODUCTION : Although patients with non-small cell lung cancer ( NSCLC ) whose tumors harbor epidermal growth factor receptor ( P00533 ) activating mutations commonly experience significant regressions when treated with erlotinib or gefitinib , they uniformly develop resistance to these agents . The secondary P00533 T790M mutation is found in 50 % of patients with acquired resistance . Herein , we studied DB05007 , an oral small molecule inhibitor of multiple receptor tyrosine kinases , including P00533 , P35968 , P04626 , and EphB4 , in NSCLC patients known or suspected of having tumors harboring T790M . METHODS : Eligible patients included those with relapsed or recurrent advanced NSCLC who progressed after ≥12 weeks of stable disease or response to erlotinib or gefitinib and/or those patients with a documented P00533 T790M . DB05007 300 mg was administered once daily . The primary end point was objective response rate . Pretreatment plasma samples were collected for mutation testing of circulating tumor DNA . RESULTS : Forty-one patients were enrolled ; 33 were evaluable for efficacy . One partial response was observed ( response rate 3 % and 90 % confidence interval , 0 % to 14 % ) . Of patients whose tumors harbored T790M , 67 % ( 8/12 ) had progression of disease as best response compared with 14 % ( 3/21 ) of those without this mutation . Plasma samples from 40 patients were available for mutation testing , 14 ( 35 % ) of which were found to have P00533 mutations . CONCLUSIONS : The 3 % response rate observed did not meet the prespecified threshold to recommend further study of DB05007 in patients who develop acquired resistance to erlotinib or gefitinib . Patients with T790M had a significantly worse progression-free survival . Retinoids induce P14780 expression through RARalpha during mammary gland remodeling . Retinoic acid ( RA ) is a signaling molecule in the morphogenesis of the mammary gland , modulating the expression of matrix metalloproteinases ( MMPs ) . The aim of this paper was to study the role of RA during weaning , which consists of three events : apoptosis of the secretory cells , degradation of the extracellular matrix , and adipogenesis . CRABP II and P09455 -1 carrier proteins increased significantly during weaning compared with lactating glands but reverted to control values after the litter resuckled . The effects of RA are mediated by the nuclear receptors RARalpha , RARbeta , RARgamma , and RXRalpha , which underwent an increase in protein levels during weaning . In an attempt to elucidate the RARalpha-dependent signaling pathway , ChIP assays were performed . The results showed the binding of RARalpha to the P14780 promoter after 24- and 72-h weaning together with its coactivator p300 ; this fact could be responsible for the increase found in P14780 mRNA and protein levels in these conditions . Expression of related MMPs ( P08253 and P08254 ) was also increased during weaning . Using gelatine zymography , we observed a time-dependent increase in active forms of P14780 and P08253 . On the other hand , the inhibitor of MMPs , P01033 , was almost undetectable at 24- and 72-h weaning by Western blot . The role of retinoids in matrix remodeling is reinforced by the fact that administration of an acute dose of retinol palmitate to control lactating rats also induces P14780 expression . This emphasizes the importance of retinoids in vivo to regulate mammary gland involution . Ligand binding domain of granulocyte colony-stimulating factor receptor . The amino-terminal domain of the cytokine receptor homologous region ( BN domain ; roughly 100 amino acid residues ) in the receptor for murine granulocyte colony-stimulating factor ( DB00099 ) was secreted as a maltose-binding protein fusion into the Escherichia coli periplasm . The murine BN domain ( mBN ) was prepared from the fusion protein by restriction protease Factor Xa digestion and purified to homogeneity . The purified BN domain specifically and stoichiometrically bound DB00099 , with an apparent dissociation constant ( Kd ) of 3-8 x 10(-8) M . The CD spectrum of the mBN domain was similar to that of the extracellular region of the human growth hormone ( GH ) receptor , which is composed of turns and beta-sheets held together by disulfide bonds . Tertiary folding and the beta-sheet of this small domain was confirmed by NMR spectroscopy . Disulfide bonds determined by peptide mapping were in the following locations : Cys107-Cys118 , Cys153-Cys162 , and Cys143-Cys194 . Among them , the first and the second produce small loops ( roughly 10 amino acid residues ) as found in the human P10912 . These results suggested that the mBN domain of the Q99062 expressed by E. coli has a P10912 -like structure . However , the third disulfide bond varied considerably between the G- P04141 and GH receptors . Disruption of these disulfide bonds in the BN domain of the Q99062 suggested that all of them are critical for maintaining a stably folded protein . Our results will facilitate understanding of the biophysical and structural properties of this receptor . DB01093 -treated HL60 cells : a model of neutrophil-like cells mainly expressing Q07343 subtype . The human promyelocytic HL60 cells acquired a neutrophilic phenotype after a 7- to 10-day DB01093 treatment . Fc gammaRII was up-regulated . Fc gammaRI was also up-regulated by an additional P01579 treatment . These cells are able to produce O2*- by NADPH oxidase activation in the presence of immune complexes or phorbol-12-myristate-13-acetate ( PMA ) . A change of their DB05876 subtype profile was also observed : Q07343 was the predominant isoenzyme , Q08499 was down-regulated and P27815 was no longer detectable . Additionally , the more NADPH oxidase was activated by PMA , the less P27815 was expressed , suggesting that NADPH oxidase activity could be used as a surrogate marker of P27815 down-regulation . DB01954 and DB03849 ( cilomilast ) , two selective DB05876 inhibitors , dose-dependently inhibited receptor-coupled activation of superoxide . These results suggest that Q07343 is the main subtype involved in regulating superoxide induced by Fc gammaRs activation . Furthermore , these cells , expressing almost exclusively Q07343 subtype , could be useful to identify selective Q07343 inhibitors . DB00559 , an endothelin receptor antagonist , ameliorates collagen-induced arthritis : the role of P01375 -α in the induction of endothelin system genes . OBJECTIVE : Endothelins ( ETs ) are involved in several inflammatory events . The present study investigated the efficacy of DB00559 , a dual P25101 /ETB receptor antagonist , in collagen-induced arthritis ( CIA ) in mice . TREATMENT : CIA was induced in DBA/1J mice . Arthritic mice were treated with DB00559 ( 100 mg/kg ) once a day , starting from the day when arthritis was clinically detectable . METHODS : CIA progression was assessed by measurements of visual clinical score , paw swelling and hypernociception . Histological changes , neutrophil infiltration and pro-inflammatory cytokines were evaluated in the joints . Gene expression in the lymph nodes of arthritic mice was evaluated by microarray technology . PreproET-1 mRNA expression in the lymph nodes of mice and in peripheral blood mononuclear cells ( PBMCs ) was evaluated by real-time PCR . The differences were evaluated by one-way Q9UNW9 or Student 's t test . RESULTS : Oral treatment with DB00559 markedly ameliorated the clinical aspects of CIA ( visual clinical score , paw swelling and hyperalgesia ) . DB00559 treatment also reduced joint damage , leukocyte infiltration and pro-inflammatory cytokine levels ( IL-1β , TNFα and Q16552 ) in the joint tissues . Changes in gene expression in the lymph nodes of arthritic mice returned to the levels of the control mice after DB00559 treatment . PreproET mRNA expression increased in PBMCs from rheumatoid arthritis ( RA ) patients but returned to basal level in PBMCs from patients under anti- P01375 therapy . In-vitro treatment of PBMCs with TNFα upregulated ET system genes . CONCLUSION : These findings indicate that ET receptor antagonists , such as DB00559 , might be useful in controlling RA . Moreover , it seems that ET mediation of arthritis is triggered by TNFα . Effect of endothelin receptor antagonists on non-muscle matrix compaction in a cell culture vasospasm model . P05305 ( ET-1 ) , a potent vascular smooth muscle constrictor , is one of the possible spasmogens in cerebral vasospasm . However , the role of ET-1 in non-muscle compaction ( another aspect of the pathogenesis of cerebral vasospasm ) has not been reported . This study was undertaken to demonstrate the effect of ET-1 , as well as erythrocyte lysate and bloody cerebrospinal fluid ( P04141 ) , on fibroblast populated collagen lattice ( FPCL ) compaction . Human dermal fibroblasts were used to form FPCL . The concentration-dependent effect of ET-1 was examined in the absence and presence of an P25101 receptor antagonist ( BQ-485 ) , or an ETB receptor antagonist ( BQ-788 ) , or both . FPCL compaction was determined by measuring reduction of areas over five days following treatment . To compare the effect of ET-1 on lattice compaction , erythrocyte lysate and bloody P04141 obtained from a cerebral vasospasm patient were also tested . We found that ET-1 increased FPCL compaction in a concentration-dependent ( but not time-dependent ) manner . Erythrocyte lysate produced the strongest compaction , however , without time-dependence . Bloody P04141 promoted FPCL compaction in a time-dependent fashion . Compaction induced by ET-1 was inhibited by BQ-485 but not by BQ-788 . We concluded that ET-1 promotes FPCL compaction by activation of P25101 receptors . Other components in bloody P04141 or erythrocytes may also contribute to FPCL compaction . Immunoregulatory natural killer cells suppress autoimmunity by down-regulating antigen-specific CD8+ T cells in mice . Natural killer ( NK ) cells belong to the innate immune system . Besides their role in antitumor immunity , NK cells also regulate the activity of other cells of the immune system , including dendritic cells , macrophages , and T cells , and may , therefore , be involved in autoimmune processes . The aim of the present study was to clarify the role of NK cells within this context . Using two mouse models for type 1 diabetes mellitus , a new subset of NK cells with regulatory function was identified . These cells were generated from conventional NK cells by incubation with Q14116 and are characterized by the expression of the surface markers CD117 ( also known as c-Kit , stem cell factor receptor ) and programmed death ( PD ) -ligand 1 . In vitro analyses demonstrated a direct lysis activity of Q14116 -stimulated NK cells against activated insulin-specific CD8(+) T cells in a P18621 /PD-ligand 1-dependent manner . Flow cytometry analyses revealed a large increase of splenic and lymphatic NK1.1(+)/c-Kit(+) NK cells in nonobese diabetic mice at 8 wk of age , the time point of acceleration of adaptive cytotoxic immunity . Adoptive transfer of unstimulated and Q14116 -stimulated NK cells into streptozotocin-treated mice led to a delayed diabetes development and partial disease prevention in the group treated with Q14116 -stimulated NK cells . Consistent with these data , mild diabetes was associated with increased numbers of NK1.1(+)/c-Kit(+) NK cells within the islets . Our results demonstrate a direct link between innate and adaptive immunity in autoimmunity with newly identified immunoregulatory NK cells displaying a potential role as immunosuppressors . Death receptors 4 and 5 activate Nox1 NADPH oxidase through riboflavin kinase to induce reactive oxygen species-mediated apoptotic cell death . Stimulation of the proapoptotic tumor necrosis factor ( P01375 ) -related apoptosis-inducing ligand ( P50591 ) receptors , death receptors 4 ( DR4 ) and 5 ( DR5 ) , conventionally induces caspase-dependent apoptosis in tumor cells . Here we report that stimulation of DR4 and/or DR5 by the agonistic protein KD548-Fc , an Fc-fused DR4/DR5 dual-specific Kringle domain variant , activates plasma membrane-associated Nox1 NADPH oxidase to generate superoxide anion and subsequently accumulates intracellular reactive oxygen species ( ROS ) , leading to sustained c-Jun N-terminal kinase activation and eventual apoptotic cell death in human HeLa and Jurkat tumor cells . KD548-Fc treatment induces the formation of a DR4/DR5 signaling complex containing riboflavin kinase ( Q969G6 ) , Nox1 , the Nox1 subunits ( Rac1 , Noxo1 , and Noxa1 ) , P01375 receptor-associated death domain ( Q15628 ) , and Q12933 ( TRAF2 ) . Depletion of Q969G6 , but not the Nox1 subunits , Q15628 and TRAF2 , failed to recruit Nox1 and Rac1 to DR4 and DR5 , demonstrating that Q969G6 plays an essential role in linking DR4/DR5 with Nox1 . Knockdown studies also reveal that Q969G6 , Q15628 , and TRAF2 play critical , intermediate , and negligible roles , respectively , in the KD548-Fc-mediated ROS accumulation and downstream signaling . Binding assays using recombinantly expressed proteins suggest that DR4/DR5 directly interact with cytosolic Q969G6 through Q969G6 -binding regions within the intracellular death domains , and Q15628 stabilizes the DR4/DR5- Q969G6 complex . Our results suggest that DR4 and DR5 have a capability to activate Nox1 by recruiting Q969G6 , resulting in ROS-mediated apoptotic cell death in tumor cells . DB05692 , a thrombin receptor ( P25116 ) antagonist for the prevention and treatment of atherothrombosis . DB05692 is a novel antiplatelet agent undergoing development by Schering-Plough Corp for the treatment and prevention of atherothrombosis . The compound is an orally administered himbacine analog that potently antagonizes the platelet thrombin receptor protease-activated receptor 1 ( P25116 ) , which leaves the procoagulant function of thrombin intact . In preclinical studies , DB05692 demonstrated no effect on bleed time or coagulation parameters . In both cynomolgus monkeys and humans , the compound had high bioavailability and inhibited ex vivo TRAP ( thrombin receptor-activating peptide ) -stimulated platelet aggregation in a potent and long-lasting manner . In a phase II clinical trial of patients undergoing percutaneous coronary intervention , DB05692 added to standard therapy with aspirin and clopidogrel did not increase major or minor thrombolysis in myocardial infarction bleeding , and demonstrated a trend toward decreased major adverse cardiovascular events versus placebo . At the time of publication , three phase III trials were underway to assess the efficacy and safety of DB05692 for at least 1 year in up to 35,000 patients with acute coronary syndromes or atherosclerosis . The distinct mechanism of action of DB05692 allows for cardiovascular protection without the liability of increased bleeding associated with other antiplatelet therapies . Phase III trials in high-risk patients will determine the use of DB05692 in cardiological practice .

MH_train_1345

MH_train_1345

MH_train_1345

interacts_with DB01109?

multiple_choice

Selective distributions of proteoglycans and their ligands in pericellular matrix of cultured fibroblasts . Implications for their roles in cell-substratum adhesion . We showed previously that a large chondroitin sulfate proteoglycan , P13611 ( also known as versican ) , inhibits cell-substratum adhesion , while basement membrane heparan sulfate proteoglycan ( recently named perlecan ) does not ( Yamagata et al. ( 1989 ) J. Biol. Chem. 264 , 8012-8018 ) . To extend our understanding of the adhesive function of these proteoglycans , we examined the pericellular localization of the proteoglycans and their ligands and also that of some matrix receptors and cytoskeletal molecules in various fibroblast culture systems . P13611 was abundant in the subcellular space of fibroblasts , but was excluded selectively from focal contacts where vinculin , integrins and fibronectin were localized . DB08818 , P16070 and tenascin were distributed similarly as P13611 . In contrast , perlecan was associated with fibronectin and was included in focal contacts . P18827 , a membrane heparan sulfate/chondroitin sulfate proteoglycan , was associated with fibronectin at the cell surface , partly at focal contacts and in association with stress fibers . Thus , complexes of P13611 with hyaluronan , tenascin and P16070 , are not involved in focal contacts . On the other hand , perlecan and syndecan-1 together with fibronectin may participate in focal contacts . The difference in localization between these proteoglycans may be related to their glycosaminoglycan content and to their distinctive roles in cell-substratum adhesion . P35354 inhibitors : better than traditional NSAIDs ? Vioxx and DB00482 may be no less risky than NSAIDs . Molecular weight and biochemical profile of a chemically modified heparin derivative , Suleparoide . Recently , a new chemically modified derivative of heparin ( Suleparoide , Syntex Laboratories Buenos Aires , Argentina ) was introduced for the prophylaxis of thrombosis and treatment of vascular disorders . This agent is claimed to contain a depolymerized , chemically modified , heparin derivative with similar biologic actions as heparan sulfate . To study the pharmacologic profile of this agent , we have defined its molecular weight distribution profile , utilizing a computerized gel permeation chromatographic system equipped with ultraviolet and refractive index detectors . Suleparoide exhibited a normal molecular distribution profile with no contaminants . It exhibited a weight average of 9.3 K DA and an apparent peak MW of 8.0 K DA . Approximately 50 % of the molecular components were < 5.0 K DA and 40 % > 5.0 K DA . The results from these studies on the mechanisms show that Suleparoide has anticoagulant activity primarily mediated through DB01109 Cofactor-II ( P05546 ) and because of its novel mechanism of action , further investigations on the biochemical profile of Suleparoide are carried out . Global clotting tests such as Activated Partial P13726 Time ( APTT ) , Heptest and Thrombin Time ( TT ) revealed a concentration dependent effect in all assays . Plasma samples supplemented with Suleparoide exhibited no significant anti-Xa and anti-IIa activities . However , in the P05546 mediated inhibitory assay for IIa , Suleparoide exhibited significant activity . In contrast , the P01008 ( DB11598 ) mediated inhibition of IIa was much weaker . DB09301 glycosaminoglycans as major P16109 ligands on metastatic breast cancer cell lines . The metastatic breast cancer cell line , 4T1 , abundantly expresses the oligosaccharide sialylated Lewis x ( sLe(x) ) . SLe(x) oligosaccharide on tumor cells can be recognized by E- and P16109 , contributing to tumor metastatic process . We observed that both selectins reacted with this cell line . However , contrary to the P16581 reactivity , which was sLe(x) dependent , P16109 reactivity with this cell line was sLe(x)-independent . The sLe(x)-Neg variant of the 4T1 cell line with markedly diminished expression of sLe(x) and lack of sLe(a) , provided a unique opportunity to characterize P16109 ligands and their contribution to metastasis in the absence of overlapping selectin ligands and P16581 binding . We observed that P16109 binding was Ca(2+)-independent and sulfation-dependent . We found that P16109 reacted primarily with cell surface chondroitin sulfate ( CS ) proteoglycans , which were abundantly and stably expressed on the surface of the 4T1 cell line . P16109 binding to the 4T1 cells was inhibited by heparin and CS glycosaminoglycans ( GAGs ) . Moreover , DB01109 administration significantly inhibited experimental lung metastasis . In addition , the data suggest that surface CS GAG chains were involved in P16109 mediated adhesion of the 4T1 cells to murine platelets and human umbilical vein endothelial cells . The data suggest that CS GAGs are also the major P16109 -reactive ligands on the surface of human MDA-MET cells . The results warrant conducting clinical studies on the involvement of cell surface CS chains in breast cancer metastasis and evaluation of various CS types and their biosynthetic pathways as target for development of treatment strategies for antimetastatic therapy of this disease . Recombinational and physical mapping of the locus for primary open-angle glaucoma ( Q99972 ) on chromosome 1q23-q25 . Primary open-angle glaucoma ( POAG ) is a leading cause of irreversible blindness in industrialized countries . A locus for juvenile-onset POAG , Q99972 , has been mapped to 1q21-q31 in a 9-cM interval . With recombinant haplotypes , we have now reduced the Q99972 interval to a maximum of 3 cM , between the D1S452/NGA1/D1S210 and NGA5 loci . These loci are 2.8 Mb apart on a 4.7-Mb contig that we have completed between the D1S2851 and D1S218 loci and that includes 96 YAC clones and 48 STSs . The new Q99972 interval itself is now covered by 25 YACs , 30 STSs , and 16 restriction enzyme site landmarks . The lack of a NotI site suggests that the region has few CpG islands and a low gene content . This is compatible with its predominant cytogenetic location on the 1q24 G-band . Finally , we have excluded important candidate genes , including genes coding for three ATPases ( P05026 , P23634 , P50993 ) , an ion channel ( VDAC4 ) , antithrombine III ( P01008 ) , and prostaglandin synthase ( P35354 ) . Our results provide a basis to identify the Q99972 gene . The angiotensin II type 1 receptor antagonist Losartan binds and activates bradykinin B2 receptor signaling . The angiotensin II type 1 receptor ( AT1R ) blocker ( ARB ) Losartan has cardioprotective effects during ischemia-reperfusion injury and inhibits reperfusion arrhythmias -effects that go beyond the benefits of lowering blood pressure . The renin-angiotensin and kallikrein-kinin systems are intricately connected and some of the cardioprotective effects of Losartan are abolished by blocking the bradykinin B2 receptor ( P30411 ) signaling . In this study , we investigated the ability of six clinically available ARBs to specifically bind and activate the P30411 . First , we investigated their ability to activate phosphoinositide ( PI ) hydrolysis in COS-7 cells transiently expressing the P30411 . We found that only Losartan activated the P30411 , working as a partial agonist compared to the endogenous ligand bradykinin . This effect was blocked by the P30411 antagonist DB06196 . A competitive binding analysis revealed that Losartan does not significantly compete with bradykinin and does not change the binding affinity of bradykinin on the P30411 . Furthermore , Losartan but not DB00796 mimicked the ability of bradykinin to increase the recovery of contractile force after metabolic stress in rat atrial tissue strips . In conclusion , Losartan is a partial agonist of the P30411 through direct binding and activation , suggesting that P30411 agonism could partly explain the beneficial effects of Losartan . Association between P16109 glycoprotein ligand-1 and pathogenesis in acute coronary syndrome assessed by optical coherence tomography . OBJECTIVE : Although monocytes appear to be actively involved in the onset of acute coronary syndrome ( ACS ) , they are heterogenous in human peripheral blood . How up-regulation of monocyte subsets leads to coronary plaque rupture followed by thrombus formation remains unclear . Recent studies have shown that P16109 glycoprotein ligand-1 ( Q14242 ) is involved in monocyte activation in patients with thrombus formation . We therefore investigated the relationship between the expression of Q14242 on monocyte subsets and thrombus formation using frequency-domain optical coherence tomography ( FD- O75051 ) in patients with ACS . METHODS : We enrolled a total of 100 individuals in this study : patients with acute myocardial infarction ( AMI , n=25 ) , unstable angina pectoris ( UAP , n=20 ) , or stable angina pectoris ( n=35 ) who underwent coronary angiography , and control subjects ( n=20 ) . Three monocyte subsets ( P08571 ++CD16- , P08571 ++CD16+ , and P08571 +CD16+ ) and the expression of Q14242 were measured by flow cytometry . In patients with AMI and UAP , FD- O75051 was performed before percutaneous coronary intervention . RESULTS : Circulating peripheral P08571 ++CD16+ monocytes expressed Q14242 more frequently than P08571 ++CD16- and P08571 +CD16+ monocytes in patients with ACS . The expression of Q14242 on circulating peripheral P08571 ++CD16+ monocytes was significantly elevated in patients with AMI compared with the other 3 groups . Moreover , the expression levels of Q14242 on P08571 ++CD16+ monocytes were significantly higher in patients with plaque rupture or intracoronary thrombus assessed by FD- O75051 . CONCLUSION : Up-regulation of Q14242 on P08571 ++CD16+ monocytes may be a crucial role in plaque rupture and thrombus formation . Expression of ras proto-oncogenes in the Dunning R3327 rat prostatic adenocarcinoma system . Steady-state levels of c-Ha-ras mRNA were measured in eight sublines of the Dunning R3327 rat prostatic adenocarcinoma . As a control , normal dorsal prostate tissue was studied . Increased expression of c-Ha-ras is associated with tumor progression in one lineage of the Dunning R3327 system ( H to AT1 to P24752 -Lu and P24752 -Ly-Lu ) . Here ras mRNA increases as the tumor advances from androgen dependence and a high degree of differentiation to an anaplastic aneuploid phenotype with high metastatic potential . However , in the other Dunning lineage ( H to HI to HI-F to P01008 ) , expression of c-Ha-ras is variable and does not correlate with tumor progression . Immunocytochemistry showed that levels of the c-Ha-ras P38936 protein paralleled steady-state mRNA levels in variants . Transfection assays , using NIH/3T3 cells , suggested that the ras loci were not activated in the R3327 tumors . Levels of P01116 mRNA were also measured in the Dunning tumors ; these did not correlate with tumor progression in either lineage . Expression of N-ras mRNA was not detected in the Dunning tumors . Identification of novel downstream molecules of tissue factor activation by comparative proteomic analysis . P13726 ( TF ) is both an initiator of blood coagulation and a signaling receptor . Using a proteomic approach , we investigated the role of TF in cell signaling when stimulated by its ligand , activated factor VII ( FVIIa ) . From a 2-D difference gel electrophoresis ( DIGE ) study we found forty one spots that were differentially regulated over time in FVIIa stimulated cells or in comparison to nonstimulated cells . Mass spectrometry identifies 23 out of these as 13 different proteins . One of them , elongation factor 2 ( P13639 ) , was investigated in greater detail by Western blot , a protein synthesis assay and cell cycle analysis . When tissue factor was stimulated by FVIIa , the phosphorylation of P13639 increased which inactivates this protein . Analyzing the effect using site inactivated FVIIa ( FVIIai ) , as well as the protease activated receptor 2 ( P55085 ) agonist SLIGKV , indicated that the inactivation was not P55085 dependent . A panel of tissue factor mutants was analyzed further to try to pinpoint what part of the cytoplasmic domain that is needed for this effect . Performing a protein synthesis assay in two different cell lines we could confirm that protein synthesis decreased upon stimulation by FVIIa . Cell cycle analysis showed that FVIIa also promotes a higher degree of cell proliferation . DB01109 's anti-inflammatory effects require glucosamine 6-O-sulfation and are mediated by blockade of L- and P-selectins . DB01109 has been used clinically as an anticoagulant and antithrombotic agent for over 60 years . Here we show that the potent anti-inflammatory property of heparin results primarily from blockade of P16109 and P14151 . DB01109 and chemically modified analogs were tested as inhibitors of selectin binding to immobilized sialyl Lewis(X) and of cell adhesion to immobilized selectins or thrombin-activated endothelial cells . Compared with unfractionated heparin , the modified heparinoids had inhibitory activity in this general order : over-O-sulfated heparin > heparin > 2-O,3-O-desulfated > or = N-desulfated/N-acetylated heparin > or = carboxyl-reduced heparin > or= N-,2-O,3-O-desulfated heparin >> 6-O-desulfated heparin . The heparinoids also showed similar differences in their ability to inhibit thioglycollate-induced peritonitis and oxazolone-induced delayed-type hypersensitivity . Mice deficient in P- or L-selectins showed impaired inflammation , which could be further reduced by heparin . However , heparin had no additional effect in mice deficient in both P- and L-selectins . We conclude that ( a ) heparin 's anti-inflammatory effects are mainly mediated by blocking P- and P14151 -initiated cell adhesion ; ( b ) the sulfate groups at P13671 on the glucosamine residues play a critical role in selectin inhibition ; and ( c ) some non-anticoagulant forms of heparin retain anti-inflammatory activity . Such analogs may prove useful as therapeutically effective inhibitors of inflammation . Inhibition of cyclin-dependent kinases , GSK-3beta and CK1 by hymenialdisine , a marine sponge constituent . BACKGROUND : Over 2000 protein kinases regulate cellular functions . Screening for inhibitors of some of these kinases has already yielded some potent and selective compounds with promising potential for the treatment of human diseases . RESULTS : The marine sponge constituent hymenialdisine is a potent inhibitor of cyclin-dependent kinases , glycogen synthase kinase-3beta and casein kinase 1 . DB02950 competes with DB00171 for binding to these kinases . A P24941 -hymenialdisine complex crystal structure shows that three hydrogen bonds link hymenialdisine to the Glu81 and Leu83 residues of P24941 , as observed with other inhibitors . DB02950 inhibits Q00535 /p35 in vivo as demonstrated by the lack of phosphorylation/down-regulation of Pak1 kinase in E18 rat cortical neurons , and also inhibits GSK-3 in vivo as shown by the inhibition of P46821 phosphorylation . DB02950 also blocks the in vivo phosphorylation of the microtubule-binding protein tau at sites that are hyperphosphorylated by GSK-3 and Q00535 /p35 in Alzheimer 's disease ( cross-reacting with Alzheimer's-specific AT100 antibodies ) . CONCLUSIONS : The natural product hymenialdisine is a new kinase inhibitor with promising potential applications for treating neurodegenerative disorders . beta-Carotene induces apoptosis and up-regulates peroxisome proliferator-activated receptor gamma expression and reactive oxygen species production in MCF-7 cancer cells . Although the pharmacological role of beta-carotene in the prevention and treatment of many cancer cells has received increasing attention , the molecular mechanisms underlying such chemopreventive activity are not clear . Since peroxisome proliferator-activated receptor gamma ( P37231 ) has been implicated in regulating breast cancer cell differentiation and apoptosis , the effects of beta-carotene on the P37231 -mediated pathway and its association with reactive oxygen species production in MCF-7 cells were investigated in the present study . The results demonstrated that beta-carotene significantly increased P37231 mRNA and protein levels in time-dependent manner . In addition , beta-carotene increased the cyclin-dependent kinase inhibitor P38936 ( P38936 /CIP1) expression and decreased the prostanoid synthesis rate-limiting enzyme cyclooxygenase-2 expression . DB07863 ( GW9662 ) , an irreversible P37231 antagonist , partly attenuated the cell death caused by beta-carotene . Further , reactive oxygen species ( ROS ) production was induced by beta-carotene , resulting in mitochondrial dysfunction and cytochrome C release . DB00143 ( DB00143 ) treatment decreases the intracellular ROS and prevents cytochrome C release and cell apoptosis induced by beta-carotene . In total , these observations suggest that the synergistic effect of P37231 expression and ROS production may account for beta-carotene-mediated anticancer activities . Site-specific mutagenesis of the histidine precursor of diphthamide in the human elongation factor-2 gene confers resistance to diphtheria toxin . Protein synthesis elongation factor 2 ( P13639 ) from eukaryotes contains a conserved post-translationally modified histidine residue known as diphthamide . DB03223 is a unique site of ADP-ribosylation by diphtheria toxin ( DT ) , which is responsible for cell killing . In this report , we describe the construction of DT-resistant HeLa cell lines by engineering the toxin-resistant form of its specific substrate , protein elongation factor-2 . Using site-specific mutagenesis of the histidine precursor of diphthamide , the histidine residue of codon 715 in human P13639 cDNA was substituted with one of four amino acid residue codons : leucine , methionine , asparagine or glutamine . Mutant EF-2s were subcloned into a pCMVexSVneo expression vector , transfected into HeLa cells , and DT-resistant cell clones were isolated . The protective effect of mutant EF-2s against cell killing by DT , after exposing all four mutant strains derived from HeLa cells to different concentrations of the toxin ( 5-20 ng/mL ) was demonstrated by : ( 1 ) the normal morphological appearance of the cells ; ( 2 ) their unaffected or slightly slower growth rates ; ( 3 ) their undisturbed electrophoretic DNA profiles whose integrity was virtually preserved . Mutant cell strains showed also considerable levels of resistance to very high concentrations of DT , in that they maintained slower but consistent rates of cell growth . It was hence concluded that despite its strict conservation and unique modification , the diphthamide histidine appears not to be essential to the function of human P13639 in protein synthesis . In addition , DT-resistant HeLa cell clones should prove valuable hosts for various DT gene-containing vectors that express the toxin intracellularly . Vitamin D analogues . The plethora of actions attributed to 1,25(OH)2D3 throughout the body have suggested potential therapeutic applications for the treatment of hyperproliferative diseases , immune dysfunction , endocrine disorders , and metabolic bone disease . However , the potent calcemic activity of the natural vitamin D hormone has precluded its use in most cases . New vitamin D analogues are under development that display greater specificity , in most cases , by retaining the therapeutic properties of 1,25(OH)2D3 , but with lower calcemic activity . Two analogues have been approved for use in patients : calcipotriol ( DB02300 from Leo Pharmaceuticals , Copenhagen , Denmark ) for the treatment of psoriasis ; and 19-nor-1,25(OH)2D2 ( DB00910 from Abbott Laboratories , Abbott Park , IL ) for secondary hyperparathyroidism . Many others analogues are currently being tested in preclinical and clinical trials for the treatment of various types of cancer and osteoporosis , and for immunosuppression . The selectivity of the analogues can be attributed to the combined interactions with four proteins : the vitamin D receptor ( P11473 ) , the serum vitamin D binding protein ( DBP ) , the vitamin D-24-hydroxylase and to a newly described membrane receptor . Low DBP affinity has been shown to be responsible for the reduced calcemic actions of calcipotriol and 22-oxacalcitriol ( O75051 ) , which is being tested for secondary hyperparathyroidism . However , the low calcemic activity of other analogues , including 19-nor-1,25(OH)2D2 , involves other , as yet undefined , mechanisms . Understanding of the molecular basis for the selectivity of the vitamin D analogues will allow the design of more effective and safer vitamin D compounds for the treatment of a wide range of clinical disorders . DB00435 and thiol redox regulation of Janus kinase activity . The activation of Janus kinases ( JAKs ) is crucial for propagation of the proliferative response initiated by many cytokines . The proliferation of various cell lines , particularly those of hematopoietic origin , is also modulated by mediators of oxidative stress such as nitric oxide and thiol redox reagents . Herein we demonstrate that nitric oxide and other thiol oxidants can inhibit the autokinase activity of rat O60674 in vitro , presumably through oxidation of crucial dithiols to disulfides within O60674 . The reduced form of O60674 is the most active form , and the oxidized O60674 form is inactive . DB00435 pretreatment of quiescent Ba/ P13726 cells also inhibits the interleukin 3-triggered in vivo activation of O60674 , a phenomenon that correlates with inhibited proliferation . Furthermore , we observed that the autokinase activity of P52333 responds in a similar fashion to thiol redox reagents in vitro and to nitric oxide donors in vivo . We suggest that the thiol redox regulation of JAKs may partially explain the generally immunosuppressive effects of nitric oxide and of other thiol oxidants . Identification of gliotropic factors that induce human stem cell migration to malignant tumor . Neural stem cells are mobile , are attracted to regions of brain damage , and can migrate a considerable distance to reach a glioma site . However , the molecular basis of the progression of gliotropism to malignant gliomas remains poorly understood . With the use of clinically and histologically assessed glioma cells , we have assessed their protein and gene profiles via proteomics and microarray approaches , and have identified candidate genes from human glioma tissues . This research is expected to provide clues to the molecular mechanisms underlying the migration of neural stem cells ( P13726 cell ) to glioma sites . The expression of 16 proteins was shown to have increased commonly in human glioma tissues . Among them , the expression of annexin A2 , P01033 , P12107 , bax , P04233 , P32971 , and O15270 were all increased in human glioma cells , as confirmed by Western blotting and immunohistochemical staining . In particular , annexin A2 effects an increase in migration toward P13726 and glioblastoma cells ( U87 cell ) in a Boyden chamber migration assay . An P29323 inhibitor ( PD98057 ) and a Q00535 inhibitor ( rescovitine ) inhibited 50 % and 90 % of annexin A2-induced migration in P13726 cells , respectively . A similar chemotactic migration was noted in P13726 and U87 cells . These results demonstrated that 7 candidate proteins may harbor a potential glioma tropism factor relevant to the pathology of malignant glioma . These results reveal that this novel molecular approach to the monitoring of glioma may provide clinically relevant information regarding tumor malignancy , and should also prove appropriate for high-throughput clinical screening applications . SAR and in vivo evaluation of 4-aryl-2-aminoalkylpyrimidines as potent and selective O60674 ( O60674 ) inhibitors . We report the discovery of a series of 4-aryl-2-aminoalkylpyrimidine derivatives as potent and selective O60674 inhibitors . High throughput screening of our in-house compound library led to the identification of hit 1 , from which optimization resulted in the discovery of highly potent and selective O60674 inhibitors . Advanced lead 10d demonstrated a significant dose-dependent pharmacodynamic and antitumor effect in a mouse xenograft model . Based upon the desirable profile of 10d ( DB05243 ) it was advanced into clinical trials . Effects of antihistamines on the function of human α7-nicotinic acetylcholine receptors . Effects of the histamine H₁ receptor ( P35367 ) antagonists ( antihistamines ) , promethazine ( PMZ ) , orphenadrine ( ORP ) , chlorpheniramine ( CLP ) , DB06691 ( PYR ) , diphenhydramine ( DPH ) , citerizine ( CTZ ) , and triprolidine ( TRP ) on the functional properties of the cloned α7 subunit of the human nicotinic acetylcholine receptor expressed in Xenopus oocytes were investigated . Antihistamines inhibited the α7-nicotinic acetylcholine receptor in the order PYR > CLP > TRP > PMZ > ORP≥DPH≥CTZ . Among the antihistamines , PYR showed the highest reversible inhibition of acetylcholine ( 100 µM ) -induced responses with IC₅₀ of 6.2 µM . PYR-induced inhibition was independent of the membrane potential and could not be reversed by increasing the concentration of acetylcholine . Specific binding of [ ¹²⁵I ] α-bungarotoxin , a selective antagonist for α7-nicotinic acetylcholine receptor , was not changed in the presence of PYR suggesting a non-competitive inhibition of nicotinic receptors . In line with functional experiments , docking studies indicated that PYR can potentially bind allosterically with the α7 transmembrane domain . Our results indicate that the H₂-H₄ receptor antagonists tested in this study ( 10 µM ) showed negligible inhibition of α7-nicotinic acetylcholine receptors . On the other hand , H₁ receptor antagonists inhibited the function of human α7-nicotinic acetylcholine receptor , with varying potencies . These results emphasize the importance of α7-nicotinic acetylcholine receptor for future pharmacological/toxicological profiling . Overexpression of bradykinin type 2 receptors on glioma cells enhances bradykinin-mediated blood-brain tumor barrier permeability increase . Variations in the expression levels of bradykinin ( BK ) type 2 receptors ( P30411 ) in different brain tumors may explain variable increases in BK-mediated blood-brain tumor barrier ( BTB ) permeability . This study investigated whether elevation of the P30411 expression levels on glioma cells enhances BK-mediated BTB permeability increases . Stable transfectants of P13671 rat glioma cells overexpressing P30411 were established by transfection with recombinant vectors harboring rat P30411 cDNA sequence . Elevated P30411 expression levels in transfectants were confirmed by quantitative real-time PCR , Western blots , and [ 3H ] -BK binding studies . BTB permeability was quantified with autoradiography and expressed as a unidirectional transport constant , Ki , for [ 14C ] -alpha-aminoisobutyric acid ( AIB : Mr 103 ) , using a rat brain tumor model . Baseline Ki values in tumors overexpressing P30411 were not significantly higher than in control tumors . Ki values after BK treatment in tumors overexpressing P30411 , however , were significantly higher than in control tumors . Western blots confirmed that P30411 expression levels in vivo in tumors overexpressing P30411 remained higher than in control tumors . These results suggested that alteration of P30411 expression levels on tumor cells could modulate BK-mediated BTB permeability . Therefore , P30411 expression levels in human glioma could be used to analyze the treatment results of patients undergoing treatment involving BK-modulated BTB permeability . [ Effect of the monophase oral contraceptive combination with 20 ug ethinyl estradiol/150 ug desogestrel on haemostasis ] . The authors examined the changes in the haemostasis during the use of the oral contraceptive combination with 20 microg ethynil estradiol/150 microg desogestrel at 35 women , a basic group , who used the oral contraceptive in the duration of 12 months and a control group ( n=35 ) , who do not use the pills . We found statistically significant increase of Antithrombin III ( P01008 ) ( p < 0.011 ) , Cofactor II of DB01109 ( HCII ) ( p < 0.001 ) , the activity of plasminogen ( p < 0.026 ) and beta2-antiplasmin ( 0.026 ) , significant decrease of P02810 ( PrC ) ( p < 0.0001 ) and of total Protein S ( TPrS ) ( p < 0.03 ) in the basic group in comparision with the control one . We do not observe significant changes in the rest of the haemostatic variables between the two groups . During the use of the oral contraceptive combination with 20 microg ethynil estradiol/150 microg desogestrel the changes in the system of the natural inhibitors are balanced by these in the system of fibrinolysis . Endogenous expression of histamine H1 receptors functionally coupled to phosphoinositide hydrolysis in P13671 glioma cells : regulation by cyclic AMP . 1. The effects of histamine receptor agonists and antagonists on phospholipid hydrolysis in rat-derived P13671 glioma cells have been investigated . 2 . P35367 -stimulation caused a concentration-dependent increase in the accumulation of total [ 3H ] -inositol phosphates in cells prelabelled with [ 3H ] -myo-inositol . The rank order of agonist potencies was histamine ( EC50 = 24 microM ) > N alpha-methylhistamine ( EC50 = 31 microM ) > 2-thiazolylethylamine ( EC50 = 91 microM ) . 3 . The response to 0.1 mM histamine was antagonized in a concentration-dependent manner by the H1-antagonists , mepyramine ( apparent Kd = 1 nM ) and (+)-chlorpheniramine ( apparent Kd = 4 nM ) . In addition , (-)-chlorpheniramine was more than two orders of magnitude less potent than its (+)-stereoisomer . 4 . Elevation of intracellular cyclic AMP accumulation with forskolin ( 10 microM , EC50 = 0.3 microM ) , isoprenaline ( 1 microM , EC50 = 4 nM ) or rolipram ( 0.5 mM ) , significantly reduced the histamine-mediated ( 0.1 mM ) inositol phosphate response by 37 % , 43 % and 26 % respectively . In contrast , 1,9-dideoxyforskolin did not increase cyclic AMP accumulation and had no effect on the phosphoinositide response to histamine . 5 . These data indicate the presence of functionally coupled , endogenous histamine H1 receptors in P13671 glioma cells . Furthermore , the results also indicate that H1 receptor-mediated phospholipid hydrolysis is inhibited by the elevation of cyclic AMP levels in these cells . Selectin ligand expression regulates the initial vascular interactions of colon carcinoma cells : the roles of CD44v and alternative sialofucosylated selectin ligands . Selectin-mediated binding of tumor cells to platelets , leukocytes , and vascular endothelium may regulate their hematogenous spread in the microvasculature . We recently reported that P16070 variant isoforms ( CD44v ) on LS174T colon carcinoma cells possess selectin binding activity . Here we extended those findings by showing that T84 and Colo205 colon carcinoma cells bind selectins via sialidase-sensitive O-linked glycans presented on CD44v , independent of heparan and chondroitin sulfate . To assess the functional role of CD44v in selectin-mediated binding , we quantified the adhesion to selectins of T84 cell subpopulations sorted based on their P16070 expression levels and stable LS174T cell lines generated using P16070 short hairpin RNA . High versus low P16070 -expressing T84 cells tethered more efficiently to P- and P14151 , but not P16581 , and rolled more slowly on P- and P16581 . Knocking down P16070 expression on LS174T cells inhibited binding to P16109 and increased rolling velocities over P- and P14151 relative to control-transfected cells , without affecting tethering and rolling on P16581 , however . Blot rolling analysis revealed the presence of alternative sialylated glycoproteins with molecular masses of approximately 170 and approximately 130 kDa , which can mediate selectin binding in P16070 -knockdown cells . DB01109 diminishes the avidity of colon carcinoma cells for P- and P14151 , which may compromise integrin-mediated firm adhesion to host cells and mitigate metastasis . Our finding that CD44v is a functional P16109 ligand on colon carcinoma provides a novel perspective on the enhanced metastatic potential associated with tumor CD44v overexpression and the role of selectins in metastasis .

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MH_train_1346

interacts_with DB09073?

multiple_choice

Fetzima ( levomilnacipran ) , a drug for major depressive disorder as a dual inhibitor for human serotonin transporters and beta-site amyloid precursor protein cleaving enzyme-1 . Pharmacological management of Major Depressive Disorder includes the use of serotonin reuptake inhibitors which targets serotonin transporters ( P31645 ) to increase the synaptic concentrations of serotonin . Beta-site amyloid precursor protein cleaving enzyme-1 ( P56817 -1 ) is responsible for amyloid β plaque formation . Hence it is an interesting target for Alzheimer 's disease ( AD ) therapy . This study describes molecular interactions of a new Food and Drug Administration approved antidepressant drug named ' Fetzima ' with P56817 -1 and P31645 . Fetzima is chemically known as levomilnacipran . The study has explored a possible link between the treatment of Depression and AD . ' Autodock 4.2 ' was used for docking study . The free energy of binding ( ΔG ) values for ' levomilnacipran- P31645 ' interaction and ' levomilnacipran- P56817 ' interaction were found to be -7.47 and -8.25 kcal/mol , respectively . DB08918 was found to interact with S438 , known to be the most important amino acid residue of serotonin binding site of P31645 during ' levomilnacipran- P31645 ' interaction . In the case of ' levomilnacipran- P56817 ' interaction , levomilnacipran interacted with two very crucial aspartic acid residues of P56817 -1 , namely , D32 and D228 . These residues are accountable for the cleavage of amyloid precursor protein and the subsequent formation of amyloid β plaques in AD brain . Hence , Fetzima ( levomilnacipran ) might act as a potent dual inhibitor of P31645 and P56817 -1 and expected to form the basis of a future dual therapy against depression and AD . It is an established fact that development of AD is associated with Major Depressive Disorder . Therefore , the design of new P56817 -1 inhibitors based on antidepressant drug scaffolds would be particularly beneficial . Induction of cytokine gene expression in human thyroid epithelial cells irradiated with HZE particles ( iron ions ) . Gene expression profiles were examined using cDNA microarray technology in human thyroid epithelial ( Htori-3 ) cells exposed to a low , non-toxic dose ( 10 cGy ) of radiation from HZE particles in the form of iron ions in the absence or presence of selenomethionine ( SeM ) . A total of 215 genes were differentially regulated 2 h after exposure to a 10-cGy dose of iron-ion radiation . In the microarray analysis , SeM had profound effects on the radiation-induced expression of several specific genes , which includes P00749 , P17936 , P15328 , P15291 and P02452 . Of particular interest to us was a gene cluster , " secreted proteins " , that was up-regulated after radiation exposure . Seven up-regulated genes of this gene cluster fall within the chemokine/cytokine gene cluster , namely , P09341 , P19875 , P05231 , P20809 , P10145 , Q13007 and TGFbeta2 . In microarray studies , the radiation-induced up-regulated expression of some these genes encoding cytokine/chemokine proteins was significantly decreased by SeM treatment . For P10145 , TGFbeta2 , P09341 and P19875 , these observations were validated by qPCR techniques . It is concluded that SeM can regulate ionizing radiation-induced gene expression and may serve as an effective countermeasure for some of the acute inflammatory/immune responses induced by low-dose HZE-particle radiation . The potential role of PD0332991 ( DB09073 ) in the treatment of multiple myeloma . INTRODUCTION : Multiple myeloma ( MM ) remains an incurable malignancy indicating a need for continued investigation of novel therapies . Recent studies have highlighted the role of cyclin-dependent kinases ( CDK ) in the pathogenesis of MM . PD0332991 ( DB09073 ) is an orally bioavailable , highly selective inhibitor of the P11802 /6-cyclin complex and downstream retinoblastoma protein ( Rb ) activation pathway that induces cell cycle arrest in the P55008 phase . AREAS COVERED : In this review , the authors summarize the role of the P11802 /6 signaling pathway in MM . They also summarize the development of PD0332991 as a specific inhibitor of P11802 /6 , and the reported preclinical and clinical data supporting the potential role of PD0332991 in MM . EXPERT OPINION : While PD0332991 is essentially cytostatic , inducing prolonged P55008 arrest , it enhances the cytotoxic effect of other agents effective in MM , including bortezomib and lenalidomide , as confirmed in early phase clinical trials . However , with a plethora of other drugs of different classes being tested in MM , further development of PD0332991 will depend on defining the most efficacious combination with least toxicity . An unexplored opportunity remains the potential protective effect of PD0332991 against lytic bone lesions of MM . The next few years are likely to better define the place of PD0332991 in the treatment of MM . Protease inhibitors prevent plasminogen-mediated , but not pemphigus vulgaris-induced , acantholysis in human epidermis . Pemphigus is an autoimmune blistering disease of the skin and mucous membranes . It is caused by autoantibodies directed against desmosomes , which are the principal adhesion structures between epidermal keratinocytes . Binding of autoantibodies leads to the destruction of desmosomes resulting in the loss of cell-cell adhesion ( acantholysis ) and epidermal blisters . The plasminogen activator system has been implicated as a proteolytic effector in pemphigus . We have tested inhibitors of the plasminogen activator system with regard to their potential to prevent pemphigus-induced cutaneous pathology . In a human split skin culture system , IgG preparations of sera from pemphigus vulgaris patients caused histopathologic changes ( acantholysis ) similar to those observed in the original pemphigus disease . All inhibitors that were tested ( active site inhibitors directed against uPA , tPA , and/or plasmin ; antibodies neutralizing the enzymatic activity of uPA or tPA ; substances interfering with the binding of uPA to its specific cell surface receptor Q03405 ) failed to prevent pemphigus vulgaris IgG-mediated acantholysis . P00747 -mediated acantholysis , however , was effectively antagonized by the synthetic active site serine protease inhibitor DB05476 or by p-aminomethylbenzoic acid . Our data argue against applying anti-plasminogen activator/anti-plasmin strategies in the management of pemphigus . Modulatory effect of phytoglycoprotein ( 38 kDa ) on cyclin D1/ P11802 in BNL CL.2 cells induced by N-methyl-N'-nitro-N-nitrosoguanidine . In the developmental stages of cancer , cell transformation occurs after the promotion stage and is a marker of cancer progression . This cell transformation is related to abnormal proliferation during the cancer initiation stage . The purpose of this study was to evaluate the effect of Styrax japonica Siebold et al . Zuccarin ( SJSZ ) glycoprotein on cell transformation in murine embryonic liver cells ( BNL CL.2 ) following N-methyl-N'-nitro-N-nitrosoguanidine ( MNNG ) treatment . To determine abnormal proliferation during the initiation stage , intracellular reactive oxygen species ( ROS ) , phosphorylation of extracellular signal-regulated kinase ( P29323 ) , p38 mitogen-activated protein kinase ( MAPK ) , activities of cell cycle-related factors [ cyclin D1/cyclin dependent kinase ( CDK ) 4 ] , cell cycle inhibitors ( p53 , P38936 , and p27 ) , nuclear factor ( NF ) -κB , and proliferating cell nuclear antigen ( P12004 ) were evaluated using Western blot analysis and real-time PCR . Our study demonstrated that SJSZ glycoprotein ( 50 μg/ml ) reduces foci formation with combined treatment [ MNNG and 12-O-tetradecanoyl phorbol-13-acetate ] of BNL CL.2 cells . With regard to proliferation-related signals , our finding indicated that SJSZ glycoprotein ( 50 μg/ml ) diminished the production of intracellular ROS , activity of phosphorylated P29323 , p38 MAPK , NF-κB ( p50 and p65 ) , P12004 , and cyclin D1/ P11802 in MNNG-induced BNL CL.2 cells . Taken together , these results lead us to speculate that SJSZ glycoprotein can inhibit abnormal cell proliferation at the initiation stage of hepatocarcinogenesis . Dedifferentiated chondrosarcoma mimicking a giant cell tumor . Is this low grade dedifferentiated chondrosarcoma ? We report a very rare case of a dedifferentiated chondrosarcoma mimicking a benign giant cell tumor . A 22-year-old male was admitted to our hospital with a history of mild left wrist pain after a skiing trauma . Radiology revealed an extensive meta-epiphyseal osteolytic lesion in the distal ulna , which appeared to be a giant cell tumor . Histological examination showed a biphasic tumor comprising chondroid and non-chondroid areas with a giant cell-rich lesion resembling a conventional giant cell tumor of the bone . Immunohistochemistry showed no expression of p16(INK4a) , P17948 , P35968 ( P35968 ) , P35916 , cKIT , Q00987 or P11802 . However , high expression of the tyrosine kinases P16234 and P09619 was observed . Molecular analysis showed no amplification of the cMYC gene and no activating mutations in the cKIT ( exons 9 and 11 ) or P16234 ( exon 18 ) genes . He has been on follow-up for ten months , with no evidence of local recurrence or metastatic disease . In summary , this report highlights a very rare case of a dedifferentiated chondrosarcoma in which the dedifferentiated component of the tumor bears histologic resemblance to a conventional giant cell tumor of bone . We suggest that this tumor might be categorized in the group of low-grade dedifferentiated chondrosarcomas . Mutational analysis of the mitochondrial P47985 of Saccharomyces cerevisiae . II . Biochemical characterization of temperature-sensitive Q13546 - mutations . Although the function of the P47985 is generally understood , little is known of how the structure of this protein supports its mechanistic role in electron transfer in the cytochrome bc1 complex . To better understand the structural basis of iron-sulfur protein function , we have undertaken a mutational analysis of the gene encoding this protein and initially isolated five temperature-sensitive iron-sulfur protein mutants ( Beckmann , J. D. , Ljungdahl , P. O. , and Trumpower , B. L. ( 1989 ) J. Biol. Chem. 264 , 3713-3722 ) . Each of the five ts-rip1- mutants exhibited pleiotropic effects . Although the mutant iron-sulfur proteins manifest several in vitro phenotypes in common , each exhibited unique characteristics . All of the ts-rip1- mutations resulted in membranes with decreased ubiquinol-cytochrome c oxidoreductase activities and decreased thermostability compared to membranes containing wild type iron-sulfur protein . All of the mutations conferred slight but significant resistance to the respiratory inhibitor myxothiazol , and one mutant was hypersensitive to inhibition by DB04799 , a structural analog of ubiquinone . In addition , one of the mutations completely blocks post-translational processing of the iron-sulfur protein , leading to accumulation of pre-iron-sulfur protein in mitochondrial membranes at nonpermissive temperatures . Finally , a mutation 12-amino acid residues away from the carboxyl terminus ( 203S ) results in an extremely unstable protein . This region of the protein may be essential in blocking degradation of pre-iron-sulfur protein by cytoplasmic proteases as the protein is imported into the mitochondria , or may be a " degradation signal , " which tags the iron-sulfur protein for turnover . Xaliproden ( SR57746A ) induces P08908 receptor-mediated Q96HU1 kinase activation in PC12 cells . Neurotrophic growth factors are involved in cell survival . However , natural growth factors have a very limited therapeutic use because of their short half-life . In the present study , we investigated the mechanism of action of a non-peptidic neurotrophic drug , Xaliproden , a potential molecule for the treatment of motoneuron diseases , since the transduction pathways of this synthetic P08908 agonist are very poorly understood . Xaliproden does not activate the Trk receptor but causes a rapid increase in the activities of the P27361 and P28482 isoforms of Q96HU1 kinase , which then rapidly decrease to the basal level . We demonstrate that isoforms of the P29353 adapter protein are phosphorylated independently of each other and are probably not the source of the Xaliproden-induced Q96HU1 kinases activation . The inhibitor of Ras farnesylation , FPT-1 , and the protein kinase C inhibitors , GF 109203X and chelerythrine , inhibited the Xaliproden-induced Q96HU1 kinase activation , suggesting p21Ras and PKC involvement . Moreover , the observations that the P08908 antagonist , pindobind , and pertussis toxin abolished the Xaliproden-induced P29323 stimulation suggested that Xaliproden activates the Q96HU1 kinase pathways by stimulating the G protein-coupled receptor , P08908 . These results demonstrate clearly that the non-peptidic compound , Xaliproden , exerts its neurotrophic effects through a mechanism of action differing from that of neurotrophins . These findings suggest that this compound does not involve MAPK activation by TrkA receptor stimulation but acts by Q96HU1 kinase pathway by a pertussis toxin-sensitive mechanism involving P08908 receptors , P38936 Ras and MEK-1 and by PKC and Akt pathways . Meningothelial hyperplasia : a detailed clinicopathologic , immunohistochemical and genetic study of 11 cases . Meningothelial hyperplasia is a poorly characterized entity , often associated with advanced age , chronic renal failure , trauma , hemorrhage , and neoplasia . In order to elucidate the nature of this lesion , 11 cases defined by the presence of nests of 10 or more cell layers thick , were compared with normal arachnoidal cap cells and meningiomas . Immunohistochemistry and Q5TCZ1 were performed to determine P35240 ( merlin ) , protein Q9Y2J2 , P15941 , progesterone receptor ( PR ) , P00533 , survivin , P15692 , DB00102 , P09619 , P12830 , and cathepsin D status . All cases had at least one putative predisposing factor , including hemorrhage ( 7 ) , chronic renal disease ( 5 ) , old age ( 5 ) , trauma ( 1 ) , and an adjacent optic nerve pilocytic astrocytoma ( 1 ) . There was typically a discontinuous growth pattern , with no invasion of surrounding normal tissue . No gene deletions were found , though scattered polyploid cells were seen in 2 cases . The immunoprofile was similar to normal cap cells with one exception ; whereas normal cells were uniformly negative for PR , nuclear positivity was seen in 64 % of hyperplasias , a frequency similar to that of benign meningiomas . Our data suggest that meningothelial hyperplasia is a reactive process that is usually distinguishable from meningioma based on clinicopathologic and genetic features . It may be preneoplastic in some , though further studies are needed to test this hypothesis . Sense p16 and antisense Q03405 bicistronic construct inhibits angiogenesis and induces glioma cell death . High-grade gliomas comprise the most malignant type of primary brain tumor and are relatively frequent in adults . Recent studies have indicated that the loss of p16 , an inhibitor of P11802 , promotes the acquisition of malignant characteristics in gliomas . A correlation between overexpression of urokinase-type plasminogen activator receptor ( Q03405 ) and glioblastoma invasion has also been established . Moreover , Q03405 /integrin binding has been shown to initiate or potentiate integrin signaling through focal adhesion kinase and/or src kinases . Our previous studies demonstrated that downregulation of Q03405 expression and restoration of p16 regress glioma growth in nude mice and downregulate alphavbeta3 integrin receptor expression . Here , we show the effect of a bicistronic construct on alphavbeta5 integrin receptor expression , angiogenesis and the biochemical pathway that causes glioma cell death . The U251 glioblastoma and a glioblastoma xenograft cell line transduced with a recombinant replication-defective adenovirus vector containing the cDNA of wild-type p16 and antisense RNA of Q03405 significantly inhibited human mammary epithelial cell capillary formation and vascular endothelial growth factor ( P15692 ) expression . Inactivation of anti-apoptotic molecules such as Akt , PARP , activation of caspases and accumulation of heteroduplex chromosomal DNA in pre- P55008 phase of the cell cycle was demonstrated by Western blotting , caspase activity assay and FACS analysis . Nuclear DNA fragmentation upon induction of apoptosis was scored using the TUNEL assay . Significant downregulation of alphavbeta5 integrin receptor expression was also confirmed by FACS analysis , immunoprecipitation and RT-PCR . Taken together , the results demonstrate that the sense p16 and anti-sense Q03405 bicistronic construct significantly inhibits angiogenesis , induces apoptosis by deregulation of the PI3K-Akt pathway and downregulates alphavbeta5 integrin receptor expression . Effects of lesopitron on the central nervous system arising from its interaction with P08908 receptors . DB04970 acts as a ligand for central serotonin P08908 receptors . Ki obtained from [3H]8-OH-DPAT competition studies was 104.8 +/- 10.6 nmol/l . As lesopitron did not affect the binding of [3H]paroxetine , involvement of the serotonin reuptake system in the effects of lesopitron is rejected . DB04970 inhibits haloperidol-induced catalepsy that is the consequence of its action on P08908 autoreceptors . The ability of lesopitron to induce 5-HT syndrome reflects post-synaptic P08908 receptor activation and the reversion of 8-OHDPAT-induced 5-HT syndrome by lesopitron suggests a partial agonist effect on this receptor-type . DB04970 induced a hypothermic effect due to the enhanced activation of post-synaptic P08908 receptors . The agonist effect of lesopitron on P08908 receptors and its marked hypothermic effect is an added value for this drug and a stimulus to the study of its possible neuroprotective action . DB05595 , an anti-folate receptor α antibody , does not block binding of folate or anti-folates to receptor nor does it alter the potency of anti-folates in vitro . PURPOSE : DB00158 is a cofactor in the synthesis of purines and pyrimidines ; folate analogs are potent cytotoxic drugs . P15328 ( FRα ) , a protein-mediating cellular accumulation of folate ( and anti-folates ) , has limited expression in normal tissues and is overexpressed by numerous carcinomas . Limited distribution and high affinity for folic acid have resulted in the development of antibodies or the use of folic acid coupled to toxins or radionuclides as therapeutic and imaging agents . DB05595 is an anti-FRα antibody in clinical trials for ovarian and non-small cell lung cancers . Our goal was to evaluate the effect of farletuzumab on binding and uptake of folates and anti-folates and the potency of anti-folates in vitro . METHODS : Direct binding and uptake of radiolabeled folates and anti-folates and the assessments of drug concentration of drug that inhibited cell growth 50 % ( IC(50) ) in vitro in the presence or absence of antibody . RESULTS : DB05595 did not block membrane binding of radiolabeled folic acid , 5-methyltetrahydrofolate , pemetrexed , and other anti-folates ; folic acid blocked > 95 % . DB05595 had a minimal effect on the cytoplasmic accumulation of 5-methyltetrahydrofolate or pemetrexed ; folic acid had a considerable but variable effect on the different cell lines . As a single agent , farletuzumab did not affect cell viability or the IC(50) of pemetrexed and other anti-folates in vitro . CONCLUSIONS : DB05595 does not block FRα binding of folates and anti-folates , minimally retards folate delivery via FRα-mediated transport , and minimally retards the growth of cells in vitro . Concomitant use of farletuzumab and pemetrexed is not contraindicated . Synthesis of oleanolic acid derivatives : In vitro , in vivo and in silico studies for P18031 inhibition . Non-insulin dependent diabetes mellitus is a multifactorial disease that links different metabolic routes ; a point of convergence is the enzyme P18031 which turns off insulin and leptin receptors involved in glucose and lipid metabolism , respectively . Pentacyclic acid triterpenes such as oleanolic acid ( OA ) have proved to be excellent P18031 inhibitors , thus , the purpose of current work was to generate a series of derivatives that improve the pharmacological effect of OA . Our findings suggest that the presence of the carboxylic acid and/or its corresponding reduction product carbinol derivative ( H-bond donor ) in C-28 is required to maintain the inhibitory activity ; moreover , this is further enhanced by ester or ether formation on C-3 . The most active derivatives were cinnamoyl ester ( 6 ) and ethyl ether ( 10 ) . DB04088 showed potent in vitro inhibitory activity and significantly decrease of blood glucose levels on in vivo experiments . Meanwhile , 10 showed contrasting outcomes , since it was the compound with higher inhibitory activity and selectivity over P18031 and has improved interaction with site B , according with docking studies , the in vivo antidiabetic effect was similar to oleanolic acid . In conclusion , oleanolic acid derivatives have revealed an enhanced inhibitory effect over P18031 activity by increasing molecular interactions with either catalytic or allosteric sites and producing a hypoglycaemic effect on non insulin dependent diabetes mellitus rat model . PPARgamma activation abolishes LDL-induced proliferation of human aortic smooth muscle cells via SOD-mediated down-regulation of superoxide . Native LDL would be a mitogenic and chemotactic stimulus of VSMC proliferation and differentiation in the atherosclerotic lesion where endothelial disruption occurred . In previous studies , our group investigated the molecular mechanisms by which LDL induces P10145 production and by which PPARalpha activation abolishes LDL effects in human aortic SMCs ( hAoSMCs ) . Herein is the first report of PPARgamma activation by troglitazone ( TG ) exerting its inhibitory effects on LDL-induced cell proliferation via generation not of H(2)O(2) , but of O2(.-) , and the subsequent activation of Erk1/2 in hAoSMCs . Moreover , in this study TG abolished the LDL-accelerated G(1)-S progression to control levels via down-regulation of active cyclinD1/ P11802 and cyclinE/ P24941 complexes and up-regulation of P38936 (Cip1) expression . TG exerted its anti-proliferative effects through the up-regulation of basal superoxide dismutase ( SOD ) expression . This data suggests that the regulation of O2(.-) is located at the crossroads between LDL signaling and cell proliferation . DB01892 , an Anti-Inflammatory Constituent from St . John 's Wort , Inhibits Microsomal Prostaglandin E(2) Synthase-1 and Suppresses Prostaglandin E(2) Formation in vivo . The acylphloroglucinol hyperforin ( Hyp ) from St . John 's wort possesses anti-inflammatory and anti-carcinogenic properties which were ascribed among others to the inhibition of P09917 . Here , we investigated whether Hyp also interferes with prostanoid generation in biological systems , particularly with key enzymes participating in prostaglandin (PG)E(2) biosynthesis , i.e. , cyclooxygenases ( P36551 ) -1/2 and microsomal PGE(2) synthase ( mPGES ) -1 which play key roles in inflammation and tumorigenesis . Similar to the mPGES-1 inhibitors MK-886 and MD-52 , Hyp significantly suppressed PGE(2) formation in whole blood assays starting at 0.03-1 μM , whereas the concomitant generation of P36551 -derived 12(S)-hydroxy-5-cis-8,10-trans-heptadecatrienoic acid , thromboxane B(2) , and 6-keto P49763 (1α) was not significantly suppressed up to 30 μM . In cell-free assays , Hyp efficiently blocked the conversion of PGH(2) to PGE(2) mediated by mPGES-1 ( IC(50) = 1 μM ) , and isolated P36551 enzymes were not ( P35354 ) or hardly ( P23219 ) suppressed . Intraperitoneal ( i.p. ) administration of Hyp ( 4 mg kg(-1) ) to rats impaired exudate volume and leukocyte numbers in carrageenan-induced pleurisy associated with reduced PGE(2) levels , and Hyp ( given i.p. ) inhibited carrageenan-induced mouse paw edema formation ( ED(50) = 1 mg kg(-1) ) being superior over indomethacin ( ED(50) = 5 mg kg(-1) ) . We conclude that the suppression of PGE(2) biosynthesis in vitro and in vivo by acting on mPGES-1 critically contributes to the anti-inflammatory efficiency of Hyp . Blastomatoid pulmonary carcinosarcoma : report of a case with a review of the literature . BACKGROUND : Pulmonary carcinosarcoma is a biphasic tumour with an unfavourable prognosis . The differential diagnosis includes pulmonary blastoma and is often challenging . CASE PRESENTATION : We here describe a case of blastomatoid pulmonary carcinosarcoma in a 58-year-old patient , who underwent surgical resection . Histopathological examination revealed immature glandular epithelium resembling high-grade fetal adenocarcinoma expressing epithelial markers and membranous beta-catenin , and blastomatoid spindle cells with partial rhabdomyosarcoma-like differentiation . Both elements expressed p53 , Q00987 , and cyclin-dependent kinase 4 ( P11802 ) , but not thyroid-transcription factor 1 ( Q15669 -1 ) . Mutation analysis of P01116 , P00533 , and beta-catenin revealed no mutations . Comparative genomic hybridization detected +1q , +6p , +6q24qter , +8q , +11q12q14 , +11q23qter , +12q12q21 , +12q24qter , +17q , +20q , -5q14q23 , -9p13pter , -13q21q21 , and amplifications at 12q14q21 , 15q24qter , 20q11q12 . CONCLUSION : The observed molecular and cytogenetic findings may provide additional tools for the differential diagnosis of biphasic pulmonary neoplasms . Furthermore , P04637 , Q00987 , P11802 , and P18031 may be involved in tumourigenesis . JTE-522 , a selective P35354 inhibitor , inhibits cell proliferation and induces apoptosis in RL95-2 cells . AIM : To investigate whether JTE-522 [ 4-(4-cyclohexyl-2-methyloxazol-5-yl)-2-fluorobenzenesulfonamide ] , a selective P35354 inhibitor , can induce apoptosis and inhibit cell proliferation in human endometrial cancer cell line RL95-2 cells and to explore the molecular mechanisms . METHODS : [ 3-(4,5)-dimethylthiazol-2-yl ] -2,5-diphenyl tetrazolium bromide ( MTT ) , DNA ladder , enzyme-linked immunosorbent assay ( ELISA ) , flow cytometry , RT-PCR , and Western blot analysis were employed to investigate effect of JTE-522 on human endometrial cancer cell line RL95-2 cells and the related molecular mechanisms . RESULTS : JTE-522 inhibited the growth of RL95-2 cells and induced the apoptosis . Furthermore , it arrested G0/ P55008 phase and inhibited S phase in RL95-2 cells . JTE-522 inhibited the expressions of P35354 mRNA , phosphorylated Rb , and P11802 proteins , while increased the levels of p53 , P38936 , cyclin D1 proteins , and the activity of caspase-3 in RL95-2 cells . CONCLUSION : JTE-522 inhibits cell proliferation and induces apoptosis in RL95-2 cells , which may be associated with the activation of caspase-3-like proteases , down-regulation of the expression of P35354 mRNA , phosphorylated Rb , and P11802 proteins , and up-regulation of the expressions of p53 , P38936 , and cyclin D1 proteins . P06401 modulator DB05366 down-regulates proliferative cell nuclear antigen and Bcl-2 protein expression and up-regulates caspase-3 and poly ( adenosine 5'-diphosphate-ribose ) polymerase expression in cultured human uterine leiomyoma cells . The present study was conducted to evaluate the effects of the progesterone receptor modulator DB05366 on proliferative activity and apoptosis in cultured human uterine leiomyoma cells . Isolated leiomyoma cells were subcultured in phenol red-free DMEM supplemented with 10 % fetal bovine serum for 120 h and then stepped down to serum-free conditions for 12 , 24 , 48 , and 96 h in the absence or presence of graded concentrations of DB05366 ( 10(-9) , 10(-8) , 10(-7) , and 10(-6) M ) . The number of viable cultured leiomyoma cells was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazodium bromide assay . P12004 ( P12004 ) expression was evaluated by immunocytochemistry and Western blot analysis . Apoptosis was examined by terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate nick end labeling ( TUNEL ) assay . P42574 , cleaved poly(ADP-ribose) polymerase ( PARP ) , and Bcl-2 expression were assessed by Western blot analysis . Compared with untreated control cultures , treatment with DB05366 decreased the number of viable cultured leiomyoma cells and the P12004 -positive rate in those cells and increased the TUNEL-positive rate in cultured leiomyoma cells in a dose-dependent manner . Western blot analysis revealed that treatment with DB05366 significantly decreased the expression of P12004 and Bcl-2 protein and increased the expression of cleaved caspase-3 and cleaved PARP in a dose-dependent manner compared with untreated control cultures . These results suggest that DB05366 inhibits the proliferation of cultured leiomyoma cells by down-regulating P12004 expression and induces apoptosis by up-regulating cleaved caspase-3 and PARP expression and down-regulating Bcl-2 protein expression in those cells . Improving molecular docking through eHiTS ' tunable scoring function . We present three complementary approaches for score-tuning that improve docking performance in pose prediction , virtual screening and binding affinity assessment . The methodology utilizes experimental data to customize the scoring function for the system of interest considering the specific docking scenario . The tuning approach , which has been implemented as an automated utility in eHiTS , is introduced as a solution to one of the conundrums of the molecular docking paradigm , namely , the lack of a universally well performing scoring function . The accuracy of scoring functions has been shown to be generally system-dependent , and particularly lacking for binding energy and bio-activity predictions . In the proposed approach , pose and energy predictions are enhanced by adjusting the relative weights of the eHiTS energy terms to improve score-RMSD or score-affinity correlations . In a virtual screening context ligand-based similarity is used to rescale the docking score such that better enrichment factors are achieved . We discuss the algorithmic details of the methods , and demonstrate the effects of score tuning on a variety of targets , including P24941 , P56817 and neuraminidase , as well as on the popular benchmarks -- the Directory of Useful Decoys and the PDBBind database . Suppressive effects of vascular endothelial growth factor-B on tumor growth in a mouse model of pancreatic neuroendocrine tumorigenesis . BACKGROUND : The family of vascular endothelial growth factors ( P15692 ) contains key regulators of blood and lymph vessel development , including P15692 , -B , -C , -D , and placental growth factor . The role of P49765 during physiological or pathological angiogenesis has not yet been conclusively delineated . Herein , we investigate the function of P49765 by the generation of mouse models of cancer with transgenic expression of P49765 or homozygous deletion of Vegfb . METHODOLOGY/PRINCIPAL FINDINGS : Ectopic expression of P49765 in the insulin-producing β-cells of the pancreas did not alter the abundance or architecture of the islets of Langerhans . The vasculature from transgenic mice exhibited a dilated morphology , but was of similar density as that of wildtype mice . Unexpectedly , we found that transgenic expression of P49765 in the Q13546 -Tag2 mouse model of pancreatic neuroendocrine tumorigenesis retarded tumor growth . Conversely , Q13546 -Tag2 mice deficient for Vegfb presented with larger tumors . No differences in vascular density , perfusion or immune cell infiltration upon altered Vegfb gene dosage were noted . However , P49765 acted to increase blood vessel diameter both in normal pancreatic islets and in Q13546 -Tag2 tumors . CONCLUSIONS/SIGNIFICANCE : Taken together , our results illustrate the differences in biological function between members of the P15692 family , and highlight the necessity of in-depth functional studies of P49765 to fully understand the effects of P17948 inhibitors currently used in the clinic .

MH_train_1347

MH_train_1347

MH_train_1347

interacts_with DB01281?

multiple_choice

The influence of costimulation and regulatory P01730 + T cells on intestinal IgA immune responses . It is thought that IgA B-cell differentiation is highly dependent on activated P01730 + T cells . In particular , cell-cell interactions in the Peyer 's patches involving P25942 and/or P33681 / P42081 have been implicated in germinal-center formation and IgA B-cell development . Also soluble factors , such as P05112 , P05113 , P05231 , and TGF beta may be critical for IgA B-cell differentiation in vivo . Here we report on some paradoxical findings with regard to IgA B-cell differentiation and specific mucosal immune responses that we have recently made using gene knockout mice . More specifically , we have investigated to what extent absence of P01730 + T cells , relevant cytokines , or T-cell-B-cell interactions would influence IgA B-cell differentiation in vivo . Using P01730 - or P05112 -gene knockout mice or mice made transgenic for DB01281 , we found that , although specific responses were impaired , total IgA production and IgA B-cell differentiation appeared to proceed normally . However , a poor correlation was found between , on the one hand , GC formation and IgA differentiation and , on the other hand , the ability to respond to T-cell-dependent soluble protein antigens in these mice . Thus , despite the various deficiencies in P01730 + T-cell functions seemingly intact IgA B-cell development was observed . P04141 -1 ( P09603 ) delivers a proatherogenic signal to human macrophages . P09603 / P09603 supports the proliferation and differentiation of monocytes and macrophages . In mice , P09603 also promotes proinflammatory responses in vivo by regulating mature macrophage functions , but little is known about the acute effects of this growth factor on mature human macrophages . Here , we show that in contrast to its effects on mouse bone marrow-derived macrophages , P09603 did not induce expression of urokinase plasminogen activator mRNA , repress expression of apolipoprotein E mRNA , or prime LPS-induced P01375 and P05231 secretion in human monocyte-derived macrophages ( HMDM ) from several independent donors . Instead , we show by expression profiling that P09603 modulates the HMDM transcriptome to favor a proatherogenic environment . P09603 induced expression of the proatherogenic chemokines P02778 /IFN-inducible protein 10 , P13500 , and P80098 but repressed expression of the antiatherogenic chemokine receptor P61073 . P09603 also up-regulated genes encoding enzymes of the cholesterol biosynthetic pathway ( P04035 , P53602 , Q13907 , P14324 , Q14534 , Q16850 , EBP , Q15738 , Q9UBM7 , and Q15392 ) , and expression of P45844 , encoding a cholesterol efflux transporter , was repressed . Consistent with these effects , P09603 increased levels of free cholesterol in HMDM , and the selective P07333 kinase inhibitor GW2580 ablated this response . These data demonstrate that P09603 represents a further link between inflammation and cardiovascular disease and suggest two distinct mechanisms by which P09603 , which is known to be present in atherosclerotic lesions , may contribute to plaque progression . Ameliorative Effect of a Selective Endothelin P25101 Receptor Antagonist in Rat Model of L- DB00134 -induced Vascular Dementia . The present study was designed to investigate the efficacy of selective P25101 receptor antagonist , ambrisentan on hyperhomocysteinemia-induced experimental vascular dementia . L-methionine was administered for 8 weeks to induce hyperhomocysteinemia and associated vascular dementia in male rats . DB06403 was administered to L-methionine-treated effect rats for 4 weeks ( starting from 5(th) to 8(th) week of L-methionine treatment ) . On 52(nd) day onward , the animals were exposed to the Morris water maze ( MWM ) for testing their learning and memory abilities . Vascular endothelial function , serum nitrite/nitrate levels , brain thiobarbituric acid reactive species ( TBARS ) , brain reduced glutathione ( DB00143 ) levels , and brain acetylcholinesterase ( P22303 ) activity were also measured . L-methionine-treated animals showed significant learning and memory impairment , endothelial dysfunction , decrease in/serum nitrite/nitrate and brain DB00143 levels along with an increase in brain TBARS levels and P22303 activity . DB06403 significantly improved hyperhomocysteinemia-induced impairment of learning , memory , endothelial dysfunction , and changes in various biochemical parameters . These effects were comparable to that of donepezil serving as positive control . It is concluded that ambrisentan , a selective P25101 receptor antagonist may be considered as a potential pharmacological agent for the management of hyperhomocysteinemia-induced vascular dementia . CCK1-receptor stimulation protects against gut mediator-induced lung damage during endotoxemia . BACKGROUND/AIMS : Cholecystokinin 1-receptor ( P32238 ) activation by long chain fatty acid ( LCFA ) absorption stimulates vago-vagal reflex pathways in the brain stem . The present study determines whether this reflex also activates the cholinergic anti-inflammatory pathway , a pathway known to modulate cytokine release during endotoxemia . METHODS : Mesenteric lymph was obtained from wild type ( WT ) and P32238 knockout ( P32238 (-/-) ) mice intraperitoneally challenged with Lipopolysaccharid ( LPS ) ( endotoxemic lymph , EL ) and intestinally infused with vehicle or LCFA-enriched solution . The lymph was analyzed for TNFα , P05231 and P22301 concentration and administered to healthy recipient mice via jugular infusion . Alveolar wall thickness , myeloperoxidase ( P05164 ) and TUNEL positive cells were determined in lung tissue of recipient mice . RESULTS : LCFA infusion in WT mice reduced TNFα concentration in EL by 49 % compared to vehicle infusion , but had no effect in P32238 (-/-) mice . EL significantly increased the alveolar wall thickness , the number of P05164 -positive and TUNEL-positive cells compared to control lymph administration . LCFA infusion in WT , but not in CCK1R(-/-) mice , significantly reduced these pathological effects of EL . CONCLUSION : During endotoxemia enteral LCFA absorption reduces TNFα release into mesenteric lymph and attenuates histomorphologic parameters of lung dysfunction . Failure to elicit this effect in CCK1R(-/-) mice demonstrates that anti-inflammatory properties of LCFAs are mediated through CCK1-Rs . P05305 induces alveolar epithelial-mesenchymal transition through endothelin type A receptor-mediated production of TGF-beta1 . P05305 ( ET-1 ) is implicated in the pathogenesis of idiopathic pulmonary fibrosis ( IPF ) , but the cellular mechanisms underlying the role it plays in this disease are not well characterized . Epithelial-mesenchymal transition ( EMT ) , which was recently demonstrated in alveolar epithelial cells ( AEC ) , may play an important role in the pathogenesis of IPF and other forms of pulmonary fibrosis . Whether ET-1 contributes to the induction of EMT in AEC is unknown . The aims of this study were to evaluate AEC production of ET-1 and to determine if ET-1 induces EMT in AEC . We demonstrate that ET-1 is produced at physiologically relevant levels by primary AEC and is secreted preferentially toward the basolateral surface . We also demonstrate that AEC express high levels of endothelin type A receptors ( P25101 ) and , to a lesser extent , type B receptors ( P24530 ) , suggesting autocrine or paracrine function for alveolar ET-1 . In addition , ET-1 induces EMT through P25101 activation . Furthermore , TGF-beta1 synthesis is increased by ET-1 , ET-1 induces P84022 phosphorylation , and ET-1-induced EMT is attenuated by a TGF-beta1-neutralizing antibody . Thus , ET-1 is an important mediator of EMT in AEC , acting through P25101 -mediated TGF-beta1 production . These findings increase our basic understanding of the role of ET-1 in pulmonary fibrosis and suggest potential roles for AEC-derived ET-1 in the pathogenesis of other alveolar epithelial-mediated lung diseases . Induction of painless thyroiditis in patients receiving programmed death 1 receptor immunotherapy for metastatic malignancies . CONTEXT : Immunotherapies against immune checkpoints that inhibit T cell activation [ cytotoxic T lymphocyte antigen 4 ( P16410 ) and programmed cell death 1 ( P18621 ) ] are emerging and promising treatments for several metastatic malignancies . However , the precise adverse effects of these therapies on thyroid gland function have not been well described . CASE DESCRIPTION : We report on 10 cases of painless thyroiditis syndrome ( Q03393 ) from a novel etiology , following immunotherapy with anti- P18621 monoclonal antibodies ( mAb ) during treatment for metastatic malignancies . Six patients presented with transient thyrotoxicosis in which thyrotropin binding inhibitory immunoglobulins ( TBII ) were absent for all , whereas four patients had evidence of positive antithyroid antibodies . All thyrotoxic patients required temporary beta-blocker therapy and had spontaneous resolution of thyrotoxicosis with subsequent hypothyroidism . Four patients presented with hypothyroidism without a detected preceding thyrotoxic phase , occurring 6-8 weeks after initial drug exposure . All of these patients had positive antithyroid antibodies and required thyroid hormone replacement therapy for a minimum of 6 months . CONCLUSIONS : Patients receiving anti- P18621 mAb therapy should be monitored for signs and symptoms of Q03393 which may require supportive treatment with beta-blockers or thyroid hormone replacement . The anti- P18621 mAb is a novel exogenous cause of Q03393 and provides new insight into the possible perturbations of the immune network that may modulate the development of endogenous Q03393 , including cases of sporadic and postpartum thyroiditis . DB03886 -dependent hyperphenylalaninemia due to deficiency of 6-pyruvoyl tetrahydropterin synthase . We describe the clinical , neurologic , and biochemical findings in 10 patients with 6-pyruvoyl tetrahydropterin synthase ( 6- Q03393 ) deficiency from seven families , all of whom originate from one large tribe in Saudi Arabia . This deficiency presents with severe , early onset of failure to thrive , neurologic deterioration , and morbidity and mortality secondary to repeated episodes of bronchopneumonia or cardiorespiratory abnormalities . The urinary pterin excretion pattern indicates deficient activity of 6- Q03393 , which has been confirmed by direct enzyme assay in red blood cells of three patients . We treated our patients with combined use of tetrahydrobiopterin 20 mg/kg/d , L-dihydroxyphenylalanine 15 mg/kg/d , carbidopa 3.75 mg/kg/d , and L-5-hydroxytryptophan 5 mg/kg/d . Neurologic findings improved significantly in all after 5 to 24 months . Although head circumference and weight returned to the lower limit of normal in four , height normalized only in one of seven patients . Despite an unrestricted diet during combined therapy , blood phenylalanine and urinary excretion of neopterin and biopterin returned to normal . DB06273 in pediatric rheumatology : the clinical experience . During the last two decades , clinical use of novel biological therapy has led to increased mechanistic understanding of complex rheumatological diseases . Conversely , basic and translational studies have led to development of new and varied therapeutic agents . These new medications which " target " specific steps in one or more immune pathways have the potential to control disease symptoms , improve quality of life and long-term prognosis , and perhaps in some , restore immunological tolerance . Use of these agents in clinical trials , combined with post-marketing surveillance , has revealed both the benefits and the undesirable side-effects of biological disease-modifying anti-rheumatic drugs ( DMARDs ) . In this review we focus on the use of tocilizumab , a monoclonal antibody directed against the P05231 receptor ( P08887 ) , which potently inhibits P05231 / P08887 signaling . P01308 -like growth factor-I inhibits transcriptional responses of transforming growth factor-beta by phosphatidylinositol 3-kinase/Akt-dependent suppression of the activation of P84022 but not Q15796 . P01308 -like growth factor-I ( P05019 ) and transforming growth factor-beta ( TGF-beta ) have been shown to be oncogenic and tumor suppressive , respectively , on prostate epithelial cells . Here we show that P05019 inhibits the ability of TGF-beta to regulate expression of several genes in the non-tumorigenic rat prostatic epithelial line , NRP-152 . In these cells , P05019 also inhibits TGF-beta-induced transcriptional responses , as shown by several promoter reporter constructs , suggesting that P05019 intercepts an early step in TGF-beta signaling . We show that P05019 does not down-regulate TGF-beta receptor levels , as determined by both receptor cross-linking and Western blot analyses . However , Western blot analysis reveals that P05019 selectively inhibits the TGF-beta-triggered activation P84022 but not Q15796 , while not altering expression of total Smads 2 , 3 , or 4 . The phosphatidylinositol 3-kinase ( PI3K ) inhibitor , LY29004 reverses the ability of P05019 to inhibit TGF-beta-induced transcriptional responses and the activation of P84022 , suggesting that the suppression of TGF-beta signaling by P05019 is mediated through activation of PI3K . Moreover , we show that enforced expression of dominant-negative PI3K ( DN-p85alpha ) or phosphatidylinositol 3-phosphate-phosphatase , P60484 , also reverse the suppressive effect of P05019 on TGF-beta-induced 3TP-luciferase reporter activity , whereas constitutively active PI3K ( p110alphaCAAX ) completely blocks TGF-beta-induced 3TP-luciferase reporter activity . Further transfection experiments including expression of constitutively active and dominant-negative Akt and rapamycin treatment suggest that suppression of TGF-beta signaling/ P84022 activation by P05019 occurs downstream of Akt and through mammalian target of rapamycin activation . In summary , our data suggest that P05019 inhibits TGF-beta transcriptional responses through selective suppression of P84022 activation via a PI3K/Akt-dependent pathway . DB01281 inhibits effector T cells through regulatory T cells and TGF-β . The P10747 costimulatory receptor is a critical regulator of T cell function , making it an attractive therapeutic target for the treatment of immune-mediated diseases . DB01281 , now approved for use in humans , prevents naive T cell activation by binding to P33681 proteins and blocking engagement of P10747 . However , DB01281 suppresses inflammation even if administered when disease is established , suggesting alternative mechanisms . We identified a novel , P10747 -independent mechanism by which DB01281 inhibits activated T cells . We show that in vitro , DB01281 synergizes with NO from bone marrow-derived macrophages to inhibit T cell proliferation . Depletion of regulatory T cells ( Tregs ) or interference with TGF-β signaling abrogated the inhibitory effect of DB01281 . Parallel in vivo experiments using an allergic airway inflammation model demonstrated that this novel mechanism required both macrophages and regulatory T cells . Furthermore , DB01281 was ineffective in P84022 -deficient mice , supporting a requirement for TGF-β signaling . Thus , in addition to preventing naive T cells from being fully activated , DB01281 can turn off already activated effector T cells by an NO/regulatory T cell/TGF-β-dependent pathway . This mechanism is similar to cell-extrinsic effects of endogenous P16410 and may be particularly important in the ability of DB01281 to treat chronic inflammatory disease . Phase I/II trial and pharmacokinetic study of cixutumumab in pediatric patients with refractory solid tumors and Ewing sarcoma : a report from the Children 's Oncology Group . PURPOSE : A phase I/II study of cixutumumab ( DB05759 ) in children with refractory solid tumors was conducted . This study was designed to assess the toxicities , pharmacokinetics , and pharmacodynamics of cixutumumab in children to determine a recommended phase II dose and to assess antitumor activity in Ewing sarcoma ( ES ) . PATIENTS AND METHODS : Pediatric patients with relapsed or refractory solid tumors were treated with cixutumumab as a 1-hour intravenous infusion once per week . Two dose levels-6 and 9 mg/kg-were evaluated using a standard three-plus-three cohort design . Patients with refractory ES were treated in an expanded phase II cohort at each dose level . RESULTS : Forty-seven eligible patients with a median age of 15 years ( range , 4 to 28 years ) were enrolled . Twelve patients were treated in the dose-finding phase . Hematologic and nonhematologic toxicities were generally mild and infrequent . Dose-limiting toxicities included grade 4 thrombocytopenia at 6 mg/kg and grade 3 dehydration at 9 mg/kg . Mean trough concentration ( ± standard deviation ) at 9 mg/kg was 106 ± 57 μg/mL , which exceeded the effective trough concentration of 60 μg/mL observed in xenograft models . Three patients with ES had confirmed partial responses : one of 10 at 6 mg/kg and two of 20 at 9 mg/kg . Serum insulin-like growth factor I ( P05019 ) levels consistently increased after one dose of cixutumumab . Tumor P08069 expression by immunohistochemistry did not correlate with response in patients with ES . CONCLUSION : Cixutumumab is well tolerated in children with refractory solid tumors . The recommended phase II dose is 9 mg/kg . Limited single-agent activity of cixutumumab was seen in ES . Possible role of cholecystokinin-A receptors in regulation of thyrotropin ( DB00024 ) secretion in male rats . We studied the importance of cholecystokinin ( CCK ) system in the regulation of thyrotropin ( DB00024 ) and prolactin ( PRL ) secretion in male rats . To this end , we tested the effects of both unselective CCK agonists CCK-8 and caerulein , and CCK-B selective agonists Q13308 and pentagastrin as well as the selective CCK antagonists ( devazepide and L-365,260 ) at wide dose-ranges on the cold-stimulated and TRH-induced DB00024 and PRL secretion . DB00403 , given s.c . 15 min before sacrifice , decreased DB00024 levels at 5 micrograms/kg . In time course-studies , the maximum inhibition was seen at 15 min but the effect lasted at least 30 , but less than 60 min . Also CCK-8 decreased DB00024 levels at the doses of 20 and 50 micrograms/kg at 15 min . Devazepide and L-365,260 did not affect DB00024 or PRL levels at any dose . The effect of caerulein ( 5 micrograms/kg ) was antagonized by devazepide , a CCK-A antagonist , at 100 micrograms/kg , but not by a CCK-B antagonist L-365,260 tested at a wide dose range . PRL levels were not affected by any treatment . DB00403 ( 5 micrograms/kg ) , given at the same time as TRH ( 500 ng/kg ) , inhibited the TRH-induced DB00024 levels at 15 min , but not at 30 or 60 min . CCK-8 ( 50 micrograms/kg ) , Q13308 ( 100 micrograms/kg ) and pentagastrin ( 500 micrograms/kg ) did not affect the TRH-induced DB00024 secretion . The results probably indicate that P32238 stimulation inhibits DB00024 secretion at the level of the anterior pituitary gland . PRL levels in male rats are not affected by CCK system . A novel inhibitory mechanism of nitrogen-containing bisphosphonate on the activity of Cl- extrusion in osteoclasts . DB09152 -containing bisphosphonates have been well known to be inhibited farnesyl diphosphate synthase ( P14324 ) , an enzyme in mevalonic acid metabolism , resulting in disturbance in polymerization of cytoskeleton structure in bone resorption and promotion of apoptosis in mature osteoclasts . Although bisphosphonates have been reported to activate ion transporters in native epithelium and Xenopus oocytes , little is known whether bisphosphonates affect acid hydrochronic acid extrusion in osteoclasts during bone resorption . The aim of this study was to determine the role of bisphosphonates on inhibition of hydrochronic acid extrusion in osteoclasts . Effects of zoledronic acid , a nitrogen-containing bisphosphonate , on the Cl(-) current activated by extracellular acidification were examined in two types of osteoclasts derived from RAW264.7 cells and mouse bone marrow macrophages ( BMMs ) . Extracellular acidification induced outwardly rectifying Cl(-) currents in mouse osteoclasts . DB00399 dose-dependently inhibited the acid-activated Cl(-) current . The non-nitrogen bisphosphonate etidronic acid had no effect on the acid-activated Cl(-) current . DB00759 -induced P14324 silencing caused a significant decrease in the Cl(-) current . The inhibitor of geranylgeranyl transferase suppressed the Cl(-) current . By contrast , the inhibitory action of zoledronic acid was rescued by addition of geranylgeranyl acid , a derivative of mevalonic acid . The activity of acid-activated Cl(-) currents was dependent on expression of P51798 during osteoclastogenesis . These results suggest that nitrogen-containing bisphosphonates suppress the activity of osteoclastic acid-activated Cl(-) currents through P14324 inhibition , suggesting the inhibition of Cl(-) extrusion activity . P62158 -mediated effects of loperamide on chloride transport by brush border membrane vesicles from human ileum . We investigated whether the synthetic opiate loperamide-HCl is able to regulate specific transport systems for sodium and chloride in brush border membrane vesicles ( BBMVs ) from human ileum and whether such activities are mediated by calcium/calmodulin . In BBMVs we studied Na+/H+ antiport , Cl+/OH- antiport , Na+/Cl- cotransport , and the Cl- conductive pathway . Brush border membrane vesicles were incubated with 10 microM loperamide over 4 h at 5 degrees C before the uptake experiments . In ileal BBMVs , loperamide stimulated intravesicular accumulation of Na+ in the presence of Cl- and vice versa . After 1 min of incubation , the stimulatory effect was 35 % +/- 5 % ( p less than 0.005 ) of the control without loperamide . DB00836 also stimulated Cl-/OH- antiport by 30 % +/- 5 % ( p less than 0.005 ) in BBMVs of ileum . In addition , we studied the role of Ca2+/calmodulin in the action of loperamide on chloride transport by human BBMVs . In loperamide-pretreated BBMVs , calmodulin activity was significantly decreased ( 12 +/- 2 vs. 38 +/- 4 pmol/mg protein ) . When loperamide-pretreated vesicles were incubated with 2 microM calcium ( free concentration ) plus 5 microM calmodulin for 1 h at 5 degrees C , complete inhibition of the stimulatory effect of loperamide on Cl-/OH- antiport and Na+/Cl- cotransport was observed . Increasing the Ca2+/calmodulin activity of loperamide-pretreated BBMVs with 2 microM calcium plus 5 microM calmodulin led to a significant inhibition of Cl-/OH- antiport and Na+/Cl- cotransport by 40 % +/- 5 % ( p less than 0.005 ) . Signal transduction by HLA class II molecules in human T cells : induction of LFA-1-dependent and independent adhesion . Crosslinking HLA-DR molecules by monoclonal antibodies ( moAbs ) induces protein tyrosine phosphorylation and results in a secondary elevation of free cytoplasmic calcium concentrations in activated human T cells . Binding of bacterial superantigens or moAbs to DR molecules on activated T cells was recently reported to induce homotypic aggregation through activation of protein kinase C ( PKC ) and mediated by CD11a/CD54 ( LFA-1/ P62158 -1 ) adhesion molecules . Here , we report that moAbs directed against framework DR , but neither DR1 , 2- and DRw52- nor DQ- and DP-specific moABs induced homotypic aggregation of antigen- and alloantigen-activated T cells , antigen-specific P01730 + T-cell lines , a CD8+ T-cytotoxic cell line , and T-leukemia cells ( HUT78 ) . Protein tyrosine kinase ( PTK ) inhibitor herbimycin A partly blocked class-II-induced aggregation responses . In contrast , phorbol ester ( PMA ) -induced aggregation was essentially unaffected . A potent inhibitor of PKC , staurosporin , inhibited both moAb- and PMA-induced aggregation responses . The aggregation responses were completely inhibited by low temperatures , cytochalasins B and E , and partly inhibited by DB00974 and P05107 moAbs , but unaffected by aphidicolin , mitomycin C , an adenylate cyclase inhibitor ( 2'5'-dideoxyadenosine ) , and moAbs against other adhesion molecules ( P06729 / P19256 [ LFA-3 ] , P10747 / P10747 ligand P33681 , P01730 , and P16070 ) . In conclusion , HLA class-II-induced aggregation responses in activated T cells appear to involve PTK and PKC activation and to be mediated through CD11a-dependent and independent adhesion pathways . Factors regulating insulin-like growth factor-binding protein-3 binding , processing , and potentiation of insulin-like growth factor action . In this study , we investigated the effects of various biochemical and pharmacological agents on insulin-like growth factor ( IGF ) -binding protein-3 ( P17936 ) cell binding and action in cultured bovine fibroblasts . When cells were preincubated for 48 h with 50 nM recombinant human ( rh ) P17936 , P05019 -stimulated [3H]aminoisobutyric acid ( [125H]AIB ) uptake was enhanced 2- to 3-fold . The addition of cytoskeletal disrupting agents during the preincubation with DB05897 did not affect P17936 potentiation of P05019 action , nor did a variety of serine , aspartate , and metalloproteinase inhibitors . On the other hand , ammonium chloride and chloroquine , weak bases that neutralize the pH of acidic cell compartments , blocked P17936 potentiation of P05019 -stimulated [3H]AIB uptake . Chloroquine and ammonium chloride had no effect alone and did not inhibit P08069 binding or action in the absence of DB05897 . Bafilomycin A , a specific inhibitor of DB00171 -dependent hydrogen ion pumps , also inhibited P17936 potentiation of P05019 -stimulated [3H]AIB uptake . Competitive [125I] P05019 binding and affinity cross-linking experiments suggested structure/function changes in cell-bound P17936 that were altered in the presence of chloroquine and bafilomycin . DB01109 markedly decreased initial P17936 cell adherence , but could not promote dissociation of P17936 from cells after the 48-h preincubation . Moreover , heparin did not inhibit P17936 potentiation of P05019 action . In summary , these data indicate that P17936 undergoes specific pH-dependent structural and/or environmental modifications that mediate the enhancing effect of P17936 on P05019 action in bovine fibroblasts . They also suggest that P17936 binding to heparin-like molecules on the cell surface is not directly involved in this process . The inhibition of inducible nitric oxide synthase reverts arthritic-induced decrease in pituitary growth hormone mRNA but not in liver insulin-like growth factor I mRNA expression . Experimental arthritis induced by Freund-adjuvant administration is a model of chronic inflammation and rheumatoid arthritis associated with a decrease in pituitary growth hormone ( GH ) and hepatic insulin-like growth factor I ( P05019 ) gene expression . Excessive nitric oxide ( NO ) synthesis by inducible NO synthase ( P35228 ) has been implicated in the pathogenesis of inflammatory illness . Moreover , NO participates in the regulation of GH secretion at both the hypothalamus and the pituitary . We have examined the role of P35228 activation in producing the changes in the GH- P05019 axis in arthritic rats . Adult male Wistar rats received aminoguanidine or vehicle from day 20 , after adjuvant or vehicle injection , until day 28 . Two hours and 30 min after the last aminoguanidine injection , all rats were killed by decapitation . Arthritis increased hypothalamic expression of somatostatin mRNA while it decreased pituitary GH mRNA expression , and both effects were prevented by aminoguanidine administration . In arthritic rats , the parallel decrease in serum P05019 , and in hepatic P05019 content and mRNA expression , correlates with the decrease in circulating GH concentrations . DB02533 administration to arthritic rats did not modify either serum GH or serum P05019 concentrations , or hepatic P05019 mRNA expression . However , aminoguanidine administration to control rats resulted in a decrease in serum GH concentrations and in a decrease in both hepatic P05019 mRNA expression and serum P05019 concentrations . These data suggest that NO mediates the arthritis-induced decrease in GH mRNA expression by acting at a hypothalamic level , but it is not involved in the decrease in hepatic P05019 mRNA expression .

MH_train_1348

MH_train_1348

MH_train_1348

interacts_with DB00316?

multiple_choice

Anti-inflammatory effect of transduced PEP-1-cyclophilin A in Raw264.7 cells and 12-O-tetradecanoylphorbol-13-acetate-induced mice . AIMS : P62937 ( CypA ) is an immunophilin that acts as a receptor for the immunosuppressant drug cyclosporine A ( DB00091 ) . CypA has emerged as a potential drug target for several inflammatory diseases , although the details of its mechanism are unclear . We examined the protective effects of CypA on inflammation in Raw 264.7 cells and animal models . MAIN METHODS : A human CypA gene was fused with a protein transduction domain , PEP-1 peptide , to construct a cell permeable PEP-1-CypA protein . The protein expression level of cyclooxygenase-2 ( P35354 ) and cytokines was detected by Western blot , ELISA and mRNA level of P35354 and cytokines were measured by RT-PCR . The nuclear factor-kappa B ( NF-kB ) and mitogen-activated protein kinase ( MAPK ) activation were analyzed by Western blot and electrophoretic mobility shift assay . Skin inflammation was detected with immunohistochemistry . KEY FINDINGS : Transduced PEP-1-CypA protein markedly inhibited lipopolysaccharide- and 12-O-tetradecanoyl phorbol-13-acetate-induced expression levels of P35354 as well as pro-inflammatory cytokine levels in vitro and in vivo . Furthermore , transduced PEP-1-CypA protein resulted in a significant reduction in the activation of NF-kB and MAPK . SIGNIFICANCE : The results indicate that PEP-1-CypA inhibits inflammatory response cytokines and enzymes by blocking NF-kB and MAPK activation upon stimulation of inflammation in vitro and in vivo . PEP-1-CypA protein may potentially be used as a therapeutic agent against skin diseases-related inflammation . 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 . Arsenic reduces the antipyretic activity of paracetamol in rats : modulation of brain P35354 activity and CB₁ receptor expression . We examined whether subacute arsenic exposure can reduce paracetamol-mediated antipyretic activity by affecting P36551 pathway and cannabinoid P21554 receptor regulation . Rats were preexposed to elemental arsenic ( 4 ppm ) as sodium arsenite through drinking water for 28 days . Next day pyrexia was induced with lipopolysaccharide and paracetamol 's ( 200 mg/kg , oral ) antipyretic activity was assessed . The activities of P23219 and P35354 , the levels of PGE₂ , P01375 -α and IL-1β and expression of CB₁ receptors were assessed in brain . Arsenic inhibited paracetamol-mediated antipyretic activity . P23219 activity was not affected by any treatments . DB00316 decreased P35354 activity , levels of PGE₂ , P01375 -α and IL-1β and caused up-regulation of P21554 receptors . Arsenic caused opposite effects on these parameters . In the arsenic-preexposed rats , paracetamol-mediated effects were attenuated , while CB₁ receptor up-regulation was reversed to down-regulation . Results suggest that elevated P35354 activity and reduced CB₁ expression could be involved in the arsenic-mediated attenuation of the antipyretic activity of paracetamol . 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 . [ Mechanism of biological actions of quercetin based on biomolecular network ] . The mechanism of biological actions of quercetin was studied by using metabolomic method and biomolecular network . HPLC-MS was used to analyze the serum metabolome in rats of blank group and quercetin administration group rats , and MS data were processed by MATLAB software . With multivariate statistical analysis of serum metabolite profiles , a clear separation among blank group and quercetin administration group was achieved , potential biomarkers were selected according to the parameters of variable importance in the projection ( P01282 ) and identified according to MS information and database retrieval . Four compounds , related enzymes , action targets and metabolic pathways had been confirmed , namely retinoic acid and RARbeta , arachidonate and P35354 , 3 , 5-diodotyrosine and P07202 , uridine diphosphate glucose and PDEs . The mechanism of quercetin enhancing ability of retinoic acid on the induction of RARbeta , activating P07202 , using as P35354 and PDEs inhibitor was approved by biomolecular network and related literatures . In this study , a mechanism of multiple biological actions of quercetin was evaluated at the level of the biomolecular network , metabolomics and biomolecular network can be used to investigate the biological effects mechanism of quercetin , which provided a new method to further revealing mechanism of drug action . In vitro effects of gonadotropin-releasing hormone ( DB00644 ) on Leydig cells of adult alpaca ( Lama pacos ) testis : P30968 immunolocalization , testosterone and prostaglandin synthesis , and cyclooxygenase activities . The main objective of this study was to examine the modulatory in vitro effects of gonadotropin-releasing hormone ( DB00644 ) on isolated Leydig cells of adult alpaca ( Lama pacos ) testis . We first evaluated the presence of P30968 ( GnRHR ) and cyclooxygenase ( P36551 ) 1 and P35354 in alpaca testis . We then studied the in vitro effects of buserelin ( DB00644 analogue ) , antide ( DB00644 antagonist ) , and buserelin plus antide or inhibitor of phospholipase C ( compound 48/80 ) and COXs ( acetylsalicylic acid ) on the production of testosterone , PGE(2) , and P49763 (2α) and on the enzymatic activities of P23219 and P35354 . Immunoreactivity for GnRHR was detected in the cytoplasm of Leydig cells and in the acrosomal region of spermatids . P23219 and P35354 immunosignals were noted in the cytoplasm of spermatogonia , spermatocytes , spermatids , Leydig cells , and Sertoli cells . Western blot analysis confirmed the GnRHR and P23219 presence in alpaca testis . The in vitro experiments showed that buserelin alone increased ( P < 0.01 ) and antide and buserelin plus acetylsalicylic acid decreased ( P < 0.01 ) testosterone and P49763 (2α) production and P23219 activity , whereas antide and compound 48/80 counteracted buserelin effects . Prostaglandin E(2) production and P35354 activity were not affected by buserelin or antide . These data suggest that DB00644 directly up-regulates testosterone production in Leydig cells of adult alpaca testis with a postreceptorial mechanism that involves P98160 , P23219 , and P49763 (2α) . P10275 abnormalities in identical twins with oligospermia . Clinical and biochemical studies . Identical twin brothers presented with oligospermia , small testes , normal male phenotypes , elevated serum luteinizing hormone levels , and normal or elevated serum testosterone levels . Both men had low to low-normal cytosol androgen receptor binding capacity in cultured fibroblasts from pubic skin biopsy specimens . Qualitative abnormalities of cellular androgen receptors were suggested by low-normal or low nuclear androgen uptake in fibroblasts from both brothers as well as abnormal thermolability and subnormal molybdate stabilization of androgen receptors from one brother . In vivo androgen sensitivity was assessed in one twin following administration of testosterone or the non-aromatizable androgen fluoxymesterone . DB01185 suppressed serum luteinizing hormone and serum testosterone/estradiol-binding globulin , and although testosterone suppressed both serum luteinizing hormone and serum follicle-stimulating hormone , the suppression of serum luteinizing hormone by testosterone was subnormal . Both subjects showed marked exaggeration of the serum 17-hydroxyprogesterone increase after administration of human chorionic gonadotropin , despite normal serum testosterone increases , suggesting a block in testicular 17,20-desmolase , which converts 17-hydroxyprogesterone to testosterone . These studies suggest that oligospermia and block of the enzyme 17,20-desmolase may be the earliest manifestations of androgen resistance , and the finding of the syndrome of oligospermia , normal male phenotype , and androgen receptor abnormalities in identical twins indicates a genetic etiology of this disorder . The relationship of in vivo central P21554 receptor occupancy to changes in cortical monoamine release and feeding elicited by P21554 receptor antagonists in rats . RATIONALE : Cannabinoid type 1 ( CB(1) ) receptor antagonists are reportedly effective in reducing food intake both preclinically and clinically . This may be due in part to their effects on monoamine release in the brain . The level of central CB(1) receptor occupancy underlying these neurobiological effects is unclear . OBJECTIVES : We explored the relationship between in vivo CB(1) receptor occupancy in the frontal cortex and changes in both monoamine release in the medial prefrontal cortex ( mPFC ) and feeding behavior in rats in response to two orally administered CB(1) receptor antagonists presently in clinical trials , SR141716A ( rimonabant ) and DB05077 . METHODS : CB(1) receptor occupancy was measured using [ (3)H ] SR141716A , and these occupancies were related to potencies to mediate increases in dopamine ( DA ) and norepinephrine ( NE ) release measured with microdialysis and decreases in consumption of a highly palatable diet ( HP ) . RESULTS : High receptor occupancy levels ( > 65 % ) were required to detect increases in monoamine release that were achieved with 3 and 10 mg/kg of SR141716A and 10 mg/kg of DB05077 for DA and 10 mg/kg of SR141716A for NE . Decreases in HP consumption were seen at occupancies higher than 65 % for SR141716A that were achieved with 3 and 10 mg/kg . In contrast , decreases in HP consumption were seen at relatively low CB(1) receptor occupancies ( 11 % ) for DB05077 . CONCLUSIONS : The occupancy method described here is an effective tool for interrelating central CB(1) receptor occupancy with neurobiological actions of CB(1) receptor antagonists and for furthering our understanding of the role of CB(1) receptors in central nervous system physiology and pathology . Design and synthesis of conformationally constrained cyclophilin inhibitors showing a cyclosporin-A phenotype in C. elegans . P62937 ( CypA ) is a member of the immunophilin family of proteins and receptor for the immunosuppressant drug cyclosporin A ( DB00091 ) . Here we describe the design and synthesis of a new class of small-molecule inhibitors for CypA that are based upon a dimedone template . Electrospray mass spectrometry is utilised as an initial screen to quantify the protein affinity of the ligands . Active inhibitors and fluorescently labelled derivatives are then used as chemical probes for investigating the biological role of cyclophilins in the nematode Caenorhabditis elegans . Tachykinin receptor modulation of cyclooxygenase-2 expression in human polymorphonuclear leucocytes . BACKGROUND AND PURPOSE : We investigated the ability of natural and synthetic selective NK receptors agonists and antagonists to modulate cyclooxygenase-2 ( P35354 ) expression in human polymorphonuclear leucocytes ( PMNs ) . EXPERIMENTAL APPROACH : The presence of all three tachykinin in PMNs was assessed by Western blot and PCR techniques . Natural and synthetic ligands selective for the tachykinin receptors were used to modulate P35354 protein ( measured with Western blotting ) and activity [ as prostaglandin E(2) ( PGE(2) ) output ] . Effects of DB05875 ( SP ) on phosphorylation of mitogen-activated protein kinases ( MAPKs ) and nuclear factor-kappa B ( NF-kappaB ) activation were studied to analyse the signalling pathway involved in P35354 up-regulation mediated by SP . KEY RESULTS : Stimulation of NK receptors with the natural ligands SP , neurokinin A ( P20366 ) and neurokinin B , in the pmol.L(-1)-micromol.L(-1) concentration range , modulated P35354 expression and PGE(2) release in a concentration- and time-dependent manner . Experiments with synthetic selective agonists [ Sar(9) , DB00134 (O(2))(11) ] SP , [ beta-Ala(8) ] P20366 (4-10) , senktide or selective antagonists L703,606 , SR48,968 or SR142801 , confirmed that P35354 up-regulation was mediated by NK receptors . We found that mainly p38 , Q8NFH3 and Q9BY77 MAPKs were phosphorylated by SP and SB202190 , PD98059 and SP600125 , which are selective inhibitors of these kinases , blocked SP-induced P35354 expression . SP also induced nuclear translocation of NF-kappaB concentration-dependently , with a maximum effect at 1 nmol.L(-1) . CONCLUSIONS AND IMPLICATIONS : Human PMNs possess functional NK(1) , NK(2) and NK(3) receptors , which mediate the induction of P35354 expression and NF-kappaB activation by SP . Altered regulation of renal nitric oxide , atrial natriuretic peptide and cyclooxygenase systems in aldosterone escape in rats . The present study was aimed to determine whether there is an altered role of local nitric oxide ( NO ) , atrial natriuretic peptide ( P01160 ) and cyclooxygenase ( P36551 ) systems in the kidney in association with the aldosterone escape . Male Sprague-Dawley rats were used . DB04630 ( 200 microg/day ) was infused through entire time course . The control group was kept on a low sodium diet ( 0.02 mEq/day ) , and the experimental group was supplied with a higher sodium diet ( 2. /day ) . Four days after beginning the regimen , the kidneys were taken . The protein expression of NO synthase ( NOS ) and P36551 isoforms was determined by semiquantitative immunoblotting . The mRNA expression of components of P01160 system was determined by real-time polymerase chain reaction . The activities of soluble and particulate guanylyl cyclases were determined by the amount of cGMP generated in responses to sodium nitroprusside and P01160 , respectively . There developed aldosterone escape in the experimental group . Accordingly , the renal content and the urinary excretion of NO increased . The expression of P29475 was increased in the inner medulla . Neither the expression of P29474 nor that of P35228 was changed . The expression and the catalytic activity of soluble guanylyl cyclase remained unaltered . The mRNA expression of P01160 was increased . Neither the expression of P16066 or P17342 nor the activity of particulate guanylyl cyclase was altered in the papilla . The protein expression of P35354 was increased in the inner medulla , while that of P23219 remained unchanged . In conclusion , the upregulation of P29475 , P01160 , and P35354 may be causally related with the aldosterone escape . Disruption of endothelial cell mitochondrial bioenergetics in lambs with increased pulmonary blood flow . AIMS : The mitochondrial dysfunction in our lamb model of congenital heart disease with increased pulmonary blood flow ( PBF ) ( Shunt ) is associated with disrupted carnitine metabolism . Our recent studies have also shown that asymmetric dimethylarginine ( DB01686 ) levels are increased in Shunt lambs and DB01686 increases the nitration of mitochondrial proteins in lamb pulmonary arterial endothelial cells ( PAEC ) in a nitric oxide synthase ( NOS ) -dependent manner . Thus , we determined whether there was a mechanistic link between endothelial nitric oxide synthase ( P29474 ) , DB01686 , and the disruption of carnitine homeostasis in PAEC . RESULTS : Exposure of PAEC to DB01686 induced the redistribution of P29474 to the mitochondria , resulting in an increase in carnitine acetyl transferase ( P43155 ) nitration and decreased P43155 activity . The resulting increase in acyl-carnitine levels resulted in mitochondrial dysfunction and the disruption of mitochondrial bioenergetics . Since the addition of L-arginine prevented these pathologic changes , we examined the effect of L-arginine supplementation on carnitine homeostasis , mitochondrial function , and nitric oxide ( NO ) signaling in Shunt lambs . We found that the treatment of Shunt lambs with L-arginine prevented the DB01686 -mediated mitochondrial redistribution of P29474 , the nitration-mediated inhibition of P43155 , and maintained carnitine homeostasis . In turn , adenosine-5'-triphosphate levels and P29474 /heat shock protein 90 interactions were preserved , and this decreased NOS uncoupling and enhanced NO generation . INNOVATION : Our data link alterations in cellular L-arginine metabolism with the disruption of mitochondrial bioenergetics and implicate altered carnitine homeostasis as a key player in this process . CONCLUSION : L-arginine supplementation may be a useful therapy to prevent the mitochondrial dysfunction involved in the pulmonary vascular alterations secondary to increased PBF . P10275 promotes the migration and invasion of upper urinary tract urothelial carcinoma cells through the upregulation of P14780 and P35354 . Dysregulated androgen receptor ( AR ) signaling is implicated in several types of tumor , including carcinomas of the prostate , breast , liver and bladder . However , the contribution of AR to the progression of upper urinary tract urothelial carcinomas ( UUTUC ) has not been fully investigated . In the present study , we demonstrated that the AR is involved in the metastasis and invasiveness of UUTUC cells . We investigated the role of the AR in UUTUC by using UUTUC-derived BFTC 909 cells . The overexpression of AR promotes the migration and invasion of BFTC 909 cells . Expression of migration/invasion-related genes was increased in BFTC 909 cells overexpressing AR determined by qPCR and western blot analyses . The results showed that AR-enhanced migration and invasion of UUTUC cells are linked to the upregulation of the matrix-degrading enzyme P14780 and cyclooxygenase ( P36551 ) -2 . Subsequently , the blocking of P14780 and P35354 signaling by inhibitors suppressed AR-enhanced cell migration and invasion . The results of the present study provide evidence for the first time of the role of AR in the motility and invasion of UUT cancer cells and support the hypothesis that the AR may play a critical role in the establishment of the invasive phenotype in urothelial neoplasia of UUT . Thus , the AR may also serve as a novel biomarker and potential therapeutic target for UUT cancer . Effects of selective P35354 and 5- P28300 inhibition on prostaglandin and leukotriene synthesis in ductal pancreatic cancer in Syrian hamster . Selective inhibition of eicosanoid synthesis seems to decrease carcinogenesis , however , the effect on liver metastasis in pancreatic cancer is still unknown . Ductal pancreatic adenocarcinoma was chemically induced by weekly injection of N-nitrosobis-2-oxopropylamine ( BOP ) in Syrian hamster . Animals received selective inhibition of cyclooxygenase-2 ( DB00482 ) and P09917 ( DB00744 ) . In week 33 , hamsters were sacrificed and incidence of pancreatic carcinomas as well as liver metastases were examined . Furthermore , size and number of liver metastases per animal were determined and concentration of PGF1alpha , DB00917 and leukotrienes was measured in hepatic and pancreatic tissue . Combined therapy ( DB00482 + DB00744 ) significantly decreased incidence , number and size of liver metastases . Furthermore extra- and intrametastatic concentration of DB00917 was reduced by this treatment in hepatic tissue . Single Cox-2-inhibition ( DB00482 ) decreased intrametastatic hepatic PGF1alpha and DB00917 concentration while PGF1alpha concentration was reduced in non-metastatic liver ( nml ) . Moreover 5- P28300 -inhibition ( DB00744 ) decreased intrametastatic DB00917 concentration as well as PGF1alpha and DB00917 in nml . In pancreatic carcinomas highest LT-concentration was found after combined treatment and this therapy group was the only one revealing a significantly higher amount of LTs in carcinomas compared to tumour-free tissue . Hepatic LT-concentration was significantly lower in the control groups than in nml of the tumour groups . Combination of Cox-2-inhibition and 5-Lox-inhibition might be a suitable adjuvant therapy to prevent liver metastasis in human ductal pancreatic adenocarcinoma . DB05332 as a treatment for immune thrombocytopenia : a review . " Immune thrombocytopenia " ( ITP ) is an autoimmune disorder that leads to peripheral destruction , as well as a decreased production of platelets . ITP most commonly presents as mild mucocutaneous bleeding . Though it is rare , the leading cause of mortality in persons with ITP is intracranial hemorrhage and those that do not respond to therapy are at increased risk . Our understanding of the pathophysiology of ITP has evolved immensely , especially over the last 60 years . The discovery of the platelet-production stimulator , thrombopoietin ( P07202 ) , lent clarity to an earlier hypothesis that inhibition of platelet production at the level of the megakaryocyte , at least in part , accounts for thrombocytopenia in adults with ITP . This facilitated the development of P07202 -based therapies to treat ITP . P40238 agonists are one of the most recent treatments to enter the landscape . Original production of a recombinant human P07202 was halted after clinical trials revealed the untoward effect of autoantibodies to the recombinant human P07202 with cross-reactivity to endogenous P07202 . Next-step development focused on stimulation of the P07202 receptor with fewer immunogenic agents . Currently , two such thrombopoietin receptor agonists , romiplostim and eltrombopag , are licensed in the USA to treat thrombocytopenia in adults with persistent or chronic ITP . Ongoing research will assess their efficacy in other immune-mediated and nonimmune-mediated primary and secondary thrombocytopenias . [ P08473 activity in the guinea pig model of asthma ] . P08473 exists in airway epithelial cells , smooth muscle , and submucosa near glands , and cleaves tachykinins to inactive metabolites , thereby reducing there effects . To study the role of enkephalinase in asthmatic response , we measured its activity in guinea pig model of asthma . When compared with the control values , the enkephalinase activity was reduced during in immediate asthmatic response ( Q92932 ) and late asthmatic response ( P10586 ) . Compared with the control values ( 100 % ) , each value was 79.7 % , 73.4 % in the trachea and 74.3 % , 55.7 % in the lung respectively . Tracheal muscle preparation taken from the control , Q92932 , and P10586 groups were made and mounted in oxygenated modified Krebs-Ringer solution . The response was monitored by isometric transducer . Concentration response curves to P20366 with or without phosphoramidon were obtained . The contractile responses of the P10586 groups were enhanced in potency and efficiency . DB02557 potentiated the P20366 induced contraction of control and the Q92932 groups but was less potent in enhancing the contractile response in the P10586 group , showing less enkephalinase activity in the P10586 . These results suggest that the enkephalinase plays an important role in P10586 . In P10586 , the enkephalinase activity may be inhibited and the responsiveness of the smooth muscle to some bronchoconstrictor , such as tachykinins , may be increased . Impact of aromatic residues within transmembrane helix 6 of the human gonadotropin-releasing hormone receptor upon agonist and antagonist binding . To investigate the impact of aromatic residues within transmembrane helix 6 ( TMH6 ) of the human gonadotropin-releasing hormone receptor ( P30968 ) on agonist and antagonist binding , residues Y(283) , Y(284) , W(289) , Y(290) , W(291) , and F(292) were exchanged to alanine and analyzed comprehensively in functional reporter gene and ligand binding assays . Whereas receptor mutants Y(283)A , Y(284)A , and W(291)A were capable of neither ligand binding nor signal transduction , mutants W(289)A , Y(290)A , and F(292)A were functional : the F(292)A mutant behaved like wild-type receptor , while mutants W(289)A and Y(290)A differentiated between agonistic and antagonistic ligands . On the basis of the high-resolution X-ray structure of bovine rhodopsin as well as available data on P30968 mutants , models for ligand-receptor interactions are proposed . The model for D- DB00150 (6)- DB00644 ( DB06825 ) binding , representing a superagonistic ligand , is in full accordance to available data . Furthermore , new interactions are proposed : pGlu(1) interacts with N(212) in transmembrane helix 5 , DB00135 (5) with Y(290) , and D- DB00150 (6) with W(289) . The binding behavior of mutants W(289)A and Y(290)A corresponds to the proposed binding model for the antagonist DB00050 . In summary , our data as presented indicate that Y(290) plays a key function in agonist but not antagonist binding . P15121 regulates high glucose-induced ectodomain shedding of tumor necrosis factor ( P01375 ) -alpha via protein kinase C-delta and P01375 converting enzyme in vascular smooth muscle cells . Chronic low-grade inflammation has emerged as a key contributor to the cardiovascular complications of diabetes , however , the mechanisms by which diabetes increases inflammation remain poorly understood . Here , we report that exposure to high glucose ( HG ) stimulates ectodomain shedding of P01375 from rat aortic smooth muscle cells in culture . Our results show that exposure to HG decreases membrane-associated P01375 . This decrease in unprocessed P01375 was prevented by the aldose reductase ( AR ) inhibitor sorbinil and AR small interference RNA . Treatment with HG , but not equimolar mannitol or 3-O-methyl glucose , resulted in phosphorylation and activation of P01375 converting enzyme ( P78536 ) ( P78536 ) , which were attenuated by sorbinil or AR-specific small interference RNA . HG-induced P78536 phosphorylation and P01375 processing were also prevented by P01375 protease inhibitor-1 , an inhibitor of P78536 . Inhibition of protein kinase C ( PKC ) -delta by rottlerin prevented HG-induced P78536 activation and the accumulation of unprocessed P01375 . Treatment with sorbinil decreased elevated levels of circulating P01375 in streptozotocin-treated diabetic rats . DB02712 treatment also decreased the expression of P01375 , matrix metalloproteinase-2 , matrix metalloproteinase-9 , and increased tissue inhibitor of metalloproteinase-3 in vascular smooth muscle cells treated with HG and in balloon-injured carotid arteries of diabetic rats . These results indicate that HG-induced P01375 shedding could be attributed to P78536 activation , which is regulated , in part , by PKC-delta and AR . Therefore , inhibition of P78536 by P01375 protease inhibitor-1 , or pharmacological inhibition of PKC-delta or AR may represent useful strategies for treating vascular inflammation associated with diabetes .

MH_train_1349

MH_train_1349

MH_train_1349

interacts_with DB00712?

multiple_choice

DB06096 , a selective P29475 inhibitor and a P28222 /1D receptor agonist , inhibits P80511 release in preclinical migraine models . BACKGROUND : DB06096 is a combined neuronal nitric oxide synthase ( P29475 ) inhibitor and 5-hydroxytryptamine 1B/1D ( P28222 /1D ) receptor agonist . Using preclinical models , we evaluated whether these two unique therapeutic principles have a synergistic effect in attenuating stimulated calcitonin gene-related peptide ( P80511 ) release , a marker of trigeminal activation . METHODS : We examined the effect of DB06096 on : ( 1 ) DB00761 - , capsaicin- and resiniferatoxin ( RTX ) -induced immunoreactive P80511 ( iCGRP ) release from isolated preparation of rat dura mater , trigeminal ganglion ( TG ) and trigeminal nucleus caudalis ( P24821 ) ; and ( 2 ) capsaicin- and electrical stimulation ( ES ) -induced middle meningeal artery ( MMA ) dilation in a rat closed-cranial window . RESULTS : DB06096 inhibited : ( 1 ) DB00761 -stimulated iCGRP release from dura mater ( % decrease mean ± SEM , lowest effective concentration ) ( 35 ± 6 % , 30 µM ) , TG ( 24 ± 11 % , 10 µM ) and P24821 ( 40 ± 8 % , 10 µM ) ; ( 2 ) capsaicin- and RTX-induced iCGRP release from dura mater ; and ( 3 ) capsaicin- and ES-induced increase in dural artery diameter ( 32 ± 5 % , 3 mg kg(-1) intravenous ( i.v. ) and 36 ± 1 % , 10 mg kg(-1) i.v. ) . CONCLUSIONS : DB06096 inhibits P80511 release from migraine-relevant cephalic tissues . Its effect is most likely mediated via a combination of P29475 -inhibition and P28222 /1D receptor agonism in dura mater while the mechanisms of action for inhibition of P80511 release from TG and P24821 have to be investigated further . Increasing cell plating density mimics an early post-LH stage in cultured bovine granulosa cells . Cultured ovarian granulosa cells are essential models to study molecular mechanisms of gene regulation during folliculogenesis . Here , we characterize primary tissue culture models for bovine granulosa cells by morphological and physiological parameters and by novel molecular luteinization markers , as transcript abundance and DNA methylation levels . The data show that : ( 1 ) collagen substrate increased the number of attached , viable cells ; ( 2 ) the expression of the key transcripts of estrogen synthesis , P11511 , could be induced and maintained in granulosa cells from small to medium but not from large follicles , whereas ( 3 ) only granulosa cells from large but not from smaller follicles were responsive to LH ; ( 4 ) serum supplementation unfavorably transformed the cellular phenotype , induced proliferation and P12004 expression , reduced or abolished the transcript abundance of steroidogenic key genes and of gonadotropin receptor genes , P05108 , P11511 , P23945 and P22888 but , however , did not increase the abundance of the luteinization-specific marker transcripts P35354 , PTX3 , P41220 and O95498 ; but ( 5 ) by increasing the plating density , estradiol production and the abundance of P11511 transcripts , in particular those derived from the main ovarian promoter P2 , were decreased concurrently leaving P2-specific DNA methylation levels unchanged , whereas progesterone secretion was stimulated and the expression of both luteinization-specific marker transcripts , P41220 and O95498 , was significantly induced . From these data , we conclude that increasing the plating density induces a different , partly complementary , physiological and gene expression profile in cultured bovine granulosa cells and drives the cells towards an early post-LH stage of luteinization , even in the absence of luteinizing agents . 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 / Q07820 expression ratio . A screen of a public expression database of myeloma patients indicates that the P10415 / Q07820 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 , 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 . DB00126 is dispensable for oxygen sensing in vivo . Prolyl-4-hydroxylation is necessary for proper structural assembly of collagens and oxygen-dependent protein stability of hypoxia-inducible transcription factors ( HIFs ) . In vitro function of HIF prolyl-4-hydroxylase domain ( P20941 ) enzymes requires oxygen and 2-oxoglutarate as cosubstrates with iron(II) and vitamin C serving as cofactors . Although vitamin C deficiency is known to cause the collagen-disassembly disease scurvy , it is unclear whether cellular oxygen sensing is similarly affected . Here , we report that vitamin C-deprived Gulo(-/-) knockout mice show normal HIF-dependent gene expression . The systemic response of Gulo(-/-) animals to inspiratory hypoxia , as measured by plasma erythropoietin levels , was similar to that of animals supplemented with vitamin C . Hypoxic HIF induction was also essentially normal under serum- and vitamin C-free cell-culture conditions , suggesting that vitamin C is not required for oxygen sensing in vivo . Glutathione was found to fully substitute for vitamin C requirement of all 3 P20941 isoforms in vitro . Consistently , glutathione also reduced HIF-1α protein levels , transactivation activity , and endogenous target gene expression in cells exposed to CoCl(2) . A Cys201Ser mutation in Q9GZT9 increased basal hydroxylation rates and conferred resistance to oxidative damage in vitro , suggesting that this surface-accessible Q9GZT9 cysteine residue is a target of antioxidative protection by vitamin C and glutathione . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . Effect of the phosphodiesterase III inhibitor amrinone on cytokine and nitric oxide production in immunostimulated J774.1 macrophages . The level of intracellular cyclic nucleotides is a regulatory factor in a variety of immune processes . Increases in intracellular cyclic AMP ( DB02527 ) and/or cyclic GMP ( cGMP ) concentration by the inhibition of phosphodiesterase have been shown to modulate the inflammatory response . DB01427 is a clinically used positive inotropic agent which elevates intracellular DB02527 and cGMP levels by selective inhibition of the phosphodiesterase III isoenzyme . In the current study , we investigated the effect of various concentrations ( 1-300 microM ) of amrinone on lipopolysaccharide-induced production of pro- and anti-inflammatory cytokines and of nitric oxide ( NO ) in vitro . In cultured murine J774.1 macrophages , 1 ng/ml-10 microg/ml of lipopolysaccharide from Escherichia coli O55: P46977 induced production of tumor necrosis factor-alpha ( P01375 ) , interleukin-10 , and nitrite ( breakdown product of NO ) . Pretreatment of cells with amrinone caused a dose-dependent suppression of P01375 production in the concentration range of 1-100 microM . Furthermore , this drug suppressed NO production in the range of 30-300 microM . Similarly to the results in the J774.1 cells , amrinone also inhibited P01375 and NO production in the range of 10-100 microM in primary rat peritoneal macrophages . At 300 microM , but not at lower concentrations , amrinone inhibited interleukin-10 production in lipopolysaccharide-treated J774.1 macrophages . Pretreatment of the macrophages with 100 and 300 microM amrinone increased the lipopolysaccharide-elicited translocation of nuclear factor-kappa B . Taken together , our results indicate that the phosphodiesterase III inhibitor amrinone modulates the activation/production of many pro- and anti-inflammatory factors in endotoxin-stimulated cells . It remains to be further investigated how such immunomodulatory effects contribute to the clinical profile of the agent . Myeloid cell-specific serine palmitoyltransferase subunit 2 haploinsufficiency reduces murine atherosclerosis . DB00133 palmitoyltransferase ( P21549 ) is the first and rate-limiting enzyme of the de novo biosynthetic pathway of sphingomyelin ( SM ) . Both P21549 and SM have been implicated in the pathogenesis of atherosclerosis , the development of which is driven by macrophages ; however , the role of P21549 in macrophage-mediated atherogenesis is unknown . To address this issue , we have analyzed macrophage inflammatory responses and reverse cholesterol transport , 2 key mediators of atherogenesis , in P21549 subunit 2-haploinsufficient ( Sptlc2(+/-) ) macrophages . We found that Sptlc2(+/-) macrophages have significantly lower SM levels in plasma membrane and lipid rafts . This reduction not only impaired inflammatory responses triggered by O00206 and its downstream NF-κB and MAPK pathways , but also enhanced reverse cholesterol transport mediated by ABC transporters . P01130 -deficient ( Ldlr(-/-) ) mice transplanted with Sptlc2(+/-) bone marrow cells exhibited significantly fewer atherosclerotic lesions after high-fat and high-cholesterol diet feeding . Additionally , Ldlr(-/-) mice with myeloid cell-specific Sptlc2 haploinsufficiency exhibited significantly less atherosclerosis than controls . These findings suggest that P21549 could be a novel therapeutic target in atherosclerosis . 1-Dehydro-[10]-gingerdione from ginger inhibits IKKβ activity for NF-κB activation and suppresses NF-κB-regulated expression of inflammatory genes . BACKGROUND AND PURPOSE : Pungent constituents of ginger ( Zingiber officinale ) have beneficial effects on inflammatory pain and arthritic swelling . However , the molecular basis for these pharmacological properties is only partially understood . Here , we investigated the molecular target of 1-dehydro-[10]-gingerdione ( D10G ) , one of the pungent constituents of ginger , that mediates its suppression of NF-κB-regulated expression of inflammatory genes linked to toll-like receptor ( TLR ) -mediated innate immunity . EXPERIMENTAL APPROACH : RAW 264.7 macrophages or primary macrophages-derived from bone marrows of C57BL/6 or C3H/HeJ mice were stimulated with the O00206 agonist LPS in the presence of D10G . Catalytic activity of inhibitory κB ( IκB ) kinase β ( IKKβ ) was determined by a kinase assay and immunoblot analysis , and the expression of inflammatory genes by RT-PCR analysis and a promoter-dependent reporter assay . KEY RESULTS : D10G directly inhibited the catalytic activity of cell-free IKKβ . Moreover , D10G irreversibly inhibited cytoplasmic IKKβ-catalysed IκBα phosphorylation in macrophages activated by TLR agonists or P01375 -α , and also IKKβ vector-elicited NF-κB transcriptional activity in these cells . These effects of D10G were abolished by substitution of the DB00151 (179) with Ala in the activation loop of IKKβ , indicating a direct interacting site of D10G . This mechanism was shown to mediate D10G-induced disruption of NF-κB activation in LPS-stimulated macrophages and the suppression of NF-κB-regulated gene expression of inducible NOS , P35354 and P05231 . CONCLUSION AND IMPLICATIONS : This study demonstrates that IKKβ is a molecular target of D10G involved in the suppression of NF-κB-regulated gene expression in LPS-activated macrophages ; this suggests D10G has therapeutic potential in NF-κB-associated inflammation and autoimmune disorders . Hsp27 regulates epithelial mesenchymal transition , metastasis , and circulating tumor cells in prostate cancer . Defining the mechanisms underlying metastatic progression of prostate cancer may lead to insights into how to decrease morbidity and mortality in this disease . An important determinant of metastasis is epithelial-to-mesenchymal transition ( EMT ) , and the mechanisms that control the process of EMT in cancer cells are still emerging . Here , we report that the molecular chaperone Hsp27 ( P04792 ) drives EMT in prostate cancer , whereas its attenuation reverses EMT and decreases cell migration , invasion , and matrix metalloproteinase activity . Mechanistically , silencing Hsp27 decreased P05231 -dependent P40763 phosphorylation , nuclear translocation , and P40763 binding to the Twist promoter , suggesting that Hsp27 is required for P05231 -mediated EMT via modulation of P40763 /Twist signaling . We observed a correlation between Hsp27 and Twist in patients with prostate cancer , with Hsp27 and Twist expression each elevated in high-grade prostate cancer tumors . Hsp27 inhibition by DB06094 , an antisense therapy currently in phase II trials , reduced tumor metastasis in a murine model of prostate cancer . More importantly , DB06094 treatment decreased the number of circulating tumor cells in patients with metastatic castration-resistant prostate cancer in a phase I clinical trial . Overall , this study defines Hsp27 as a critical regulator of P05231 -dependent and P05231 -independent EMT , validating this chaperone as a therapeutic target to treat metastatic prostate cancer . Haploinsufficiency of the follicle-stimulating hormone receptor accelerates oocyte loss inducing early reproductive senescence and biological aging in mice . Female mice that are null for the DB00094 -receptor ( P23945 ) gene are estrogen deficient , acyclic , and sterile . However , the heterozygous ( +/- ) mice initially have reduced fertility and stop breeding by 7-9 mo . The purpose of this study was to understand the basis of reduced fertility in mice with haploinsufficiency of the P23945 . Heterozygous females were compared to +/+ females at 3 , 7 , and 12 mo of age . By 7 mo most of the +/- females were acyclic and < 50 % delivered pups . The wild-type females were normal in these respects . None of the 1-yr-old +/- females gave viable offspring ( 73 % in +/+ ) . Many degenerative changes , including atresia and apoptosis , and profound loss of oocytes , were apparent in +/- mice by 7 mo . The 1-yr-old +/- ovary had very few follicles and consisted mostly of fibroid tissue and cysts . Our data support the hypothesis that reproductive deficits in +/- P23945 mice occur because of accelerated oocyte loss due to increased cell death in the ovary . These events contribute to early reproductive senescence and biological aging in mice . Thus P23945 status is an important determinant of ovarian aging and all phenomena that arise from subsequent estrogen deficiency and other aberrations . Induction of retinoic acid receptor-beta suppresses cyclooxygenase-2 expression in esophageal cancer cells . Since retinoic acid receptor ( RAR ) -beta mRNA is frequently lost during esophageal carcinogenesis and esophageal cancer cells that do not express P10826 are resistant to retinoic acid ( RA ) , we stably transfected P10826 expression vector into an esophageal cancer cell line TE-8 and an antisense P10826 into TE-3 cells . Transfection of P10826 decreased cell growth and colony formation and induced apoptosis in TE-8 cells . Antisense P10826 -transfected TE-3 cells had a shorter doubling time and became resistant to RA . Induction of P10826 decreased P35354 expression in P10826 transfected TE-8 cells , whereas antisense P10826 transfected TE-3 cells increased P35354 expression . The inhibitory effect of P10826 on P35354 expression was further enhanced in the presence of RA , which was blocked by an RAR antagonist . The synthetic retinoid N-(4-hydroxyphenyl)retinamide , which does not bind effectively to P10826 , had no effect on P35354 suppression . Furthermore , RA blocked bile acid-induced P35354 expression and prostaglandin E(2) production only in the P10826 positive cells . Our data demonstrated that anticancer effect of P10826 may be related to its ability to suppress P35354 expression and support that the loss of P10826 expression may contribute to esophageal carcinogenesis . Gastroprotective action of orexin-A against stress-induced gastric damage is mediated by endogenous prostaglandins , sensory afferent neuropeptides and nitric oxide . O43612 -A , identified in the neurons and endocrine cells in the gut , has been implicated in control of food intake and sleep behavior but little is known about its influence on gastric secretion and mucosal integrity . The effects of orexin-A on gastric secretion and gastric lesions induced in rats by 3.5 h of water immersion and restraint stress ( P23381 ) or 75 % ethanol were determined . O43612 -A ( 5-80 microg/kg i.p. ) increased gastric acid secretion and attenuated gastric lesions induced by P23381 and this was accompanied by the significant rise in plasma orexin-A , P80511 and gastrin levels , the gastric mucosal blood flow ( GBF ) , luminal NO concentration and an increase in mRNA for P80511 and overexpression of P35354 protein and the generation of PGE(2) in the gastric mucosa . O43612 -A-induced protection was abolished by selective OX-1 receptor antagonist , vagotomy and attenuated by suppression of P23219 and P35354 , deactivation of afferent nerves with neurotoxic dose of capsaicin , pretreatment with CCK(2)/gastrin antagonist , P80511 (8-37) or capsazepine and by inhibition of NOS with DB04223 . This study shows for the first time that orexin-A exerts a potent protective action on the stomach of rats exposed to non-topical ulcerogens such as P23381 or topical noxious agents such as ethanol and these effects depend upon hyperemia mediated by P36551 -PG and NOS-NO systems , activation of vagal nerves and sensory neuropeptides such as P80511 released from sensory nerves probably triggered by an increase in gastric acid secretion induced by this peptide . Novel marine phenazines as potential cancer chemopreventive and anti-inflammatory agents . Two new ( 1 and 2 ) and one known phenazine derivative ( lavanducyanin , 3 ) were isolated and identified from the fermentation broth of a marine-derived Streptomyces sp . ( strain CNS284 ) . In mammalian cell culture studies , compounds 1 , 2 and 3 inhibited P01375 -α-induced NFκB activity ( IC₅₀ values of 4.1 , 24.2 , and 16.3 μM , respectively ) and LPS-induced nitric oxide production ( IC₅₀ values of > 48.6 , 15.1 , and 8.0 μM , respectively ) . PGE₂ production was blocked with greater efficacy ( IC₅₀ values of 7.5 , 0.89 , and 0.63 μM , respectively ) , possibly due to inhibition of cyclooxygenases in addition to the expression of P35354 . Treatment of cultured HL-60 cells led to dose-dependent accumulation in the subG1 compartment of the cell cycle , as a result of apoptosis . These data provide greater insight on the biological potential of phenazine derivatives , and some guidance on how various substituents may alter potential anti-inflammatory and anti-cancer effects . Multifaceted link between cancer and inflammation . Increasing evidence from epidemiological , preclinical and clinical studies suggests that dysregulated inflammatory response plays a pivotal role in a multitude of chronic ailments including cancer . The molecular mechanism(s) by which chronic inflammation drives cancer initiation and promotion include increased production of pro-inflammatory mediators , such as cytokines , chemokines , reactive oxygen intermediates , increased expression of oncogenes , P35354 ( cyclo-oxygenase-2 ) , 5- P28300 ( P09917 ) and MMPs ( matrix metalloproteinases ) , and pro-inflammatory transcription factors such as NF-κB ( nuclear factor κB ) , P40763 ( signal transducer and activator of transcription 3 ) , AP-1 ( activator protein 1 ) and HIF-1α ( hypoxia-inducible factor 1α ) that mediate tumour cell proliferation , transformation , metastasis , survival , invasion , angiogenesis , chemoresistance and radioresistance . These inflammation-associated molecules are activated by a number of environmental and lifestyle-related factors including infectious agents , tobacco , stress , diet , obesity and alcohol , which together are thought to drive as much as 90 % of all cancers . The present review will focus primarily on the role of various inflammatory intermediates responsible for tumour initiation and progression , and discuss in detail the critical link between inflammation and cancer . Biological differences between in vitro produced bovine embryos and parthenotes . Parthenotes may represent an alternate ethical source of stem cells , once biological differences between parthenotes and embryos can be understood . In this study , we analyzed development , trophectoderm ( TE ) differentiation , apoptosis/necrosis , and ploidy in parthenotes and in vitro produced bovine embryos . Subsequently , using real-time PCR , we analyzed the expression of genes expected to underlie the observed differences at the blastocyst stage . In vitro matured oocytes were either fertilized or activated with ionomycin +6-DMAP and cultured in simple medium . Parthenotes showed enhanced blastocyst development and diploidy and reduced TE cell counts . Apoptotic and necrotic indexes did not vary , but parthenotes evidenced a higher relative proportion of apoptotic cells between inner cell mass and TE . The pluripotence-related Q01860 and the methylation Q9Y6K1 genes were downregulated in parthenotes . Among pregnancy recognition genes , TP-1 was upregulated in parthenotes , while O00264 and PLAC8 did not change . Expression of p66(shc) and Q07812 / P10415 ratio were higher , and p53 lower , in parthenotes . Among metabolism genes , P11166 was downregulated , while P15121 , P35354 , O95479 , and P10599 were upregulated in parthenotes , and P22732 did not differ . Among genes involved in compaction/blastulation , P17302 was downregulated in parthenotes , but no differences were detected within P05023 and CDH1 . Within parthenotes , the expression levels of P11166 , TP-1 , and O95479 , and possibly P15121 , resemble patterns described in female embryos . The pro-apoptotic profile is more pronounced in parthenotes than in embryos , which may differ in their way to channel apoptotic stimuli , through p66(shc) and p53 respectively , and in their mechanisms to control pluripotency and de novo methylation . P00734 kringle-2 induces death of mesencephalic dopaminergic neurons in vivo and in vitro via microglial activation . We have shown that prothrombin kringle-2 ( pKr-2 ) , a domain of human prothrombin distinct from thrombin could activate cultured rat brain microglia in vitro . However , little is known whether pKr-2-induced microglial activation could cause neurotoxicity on dopaminergic ( DA ) neurons in vivo . To address this question , pKr-2 was injected into the rat substantia nigra ( SN ) . Tyrosine hydroxylase ( TH ) immunohistochemistry experiments demonstrate significant loss of DA neurons seven days after injection of pKr-2 . In parallel , pKr-2-activated microglia were detected in the SN with OX-42 and OX-6 immunohistochemistry . Reverse transcription PCR and double-label immunohistochemistry revealed that activated microglia in vivo exhibit early and transient expression of inducible nitric oxide synthase ( P35228 ) , cyclooxygenase-2 ( P35354 ) and several proinflammatory cytokines . The pKr-2-induced loss of SN DA neurons was partially inhibited by the NOS inhibitor N(G)-nitro-L-arginine methyl ester hydrochloride , and the P35354 inhibitor DuP-697 . P27361 /2 , c-Jun N-terminal kinase and p38 mitogen-activated protein kinase were activated in the SN as early as 1 hr after pKr-2 injection , and localized within microglia . Inhibition of these kinases led to attenuation of mRNA expression of P35228 , P35354 and several proinflammatory cytokines , and rescue of DA neurons in the SN . Intriguingly , following treatment with pKr-2 in vitro , neurotoxicity was detected exclusively in co-cultures of mesencephalic neurons and microglia , but not microglia-free neuron-enriched mesencephalic cultures , indicating that microglia are required for pKr-2 neurotoxicity . Our results strongly suggest that microglia activated by endogenous compound(s) , such as pKr-2 , are implicated in the DA neuronal cell death in the SN . Silencing of HIF prolyl-hydroxylase 2 gene in the renal medulla attenuates salt-sensitive hypertension in Dahl S rats . BACKGROUND : In response to high salt intake , transcription factor hypoxia-inducible factor ( HIF ) 1α activates many antihypertensive genes , such as heme oxygenase 1 ( P09601 ) 1 and cyclooxygenase 2 ( P35354 ) in the renal medulla , which is an important molecular adaptation to promote extra sodium excretion . We recently showed that high salt inhibited the expression of HIF prolyl-hydroxylase 2 ( Q9GZT9 ) , an enzyme that promotes the degradation of HIF-1α , thereby upregulating HIF-1α , and that high salt-induced inhibition in Q9GZT9 and subsequent activation of HIF-1α in the renal medulla was blunted in Dahl salt-sensitive hypertensive rats . This study tested the hypothesis that silencing the Q9GZT9 gene to increase HIF-1α levels in the renal medulla attenuates salt-sensitive hypertension in Dahl S rats . METHODS : Q9GZT9 short hairpin RNA ( shRNA ) plasmids were transfected into the renal medulla in uninephrectomized Dahl S rats . Renal function and blood pressure were then measured . RESULTS : Q9GZT9 shRNA reduced Q9GZT9 levels by > 60 % and significantly increased HIF-1α protein levels and the expression of HIF-1α target genes P09601 and P35354 by > 3-fold in the renal medulla . Functionally , pressure natriuresis was remarkably enhanced , urinary sodium excretion was doubled after acute intravenous sodium loading , and chronic high salt-induced sodium retention was remarkably decreased , and as a result , salt-sensitive hypertension was significantly attenuated in Q9GZT9 shRNA rats compared with control rats . CONCLUSIONS : Impaired Q9GZT9 response to high salt intake in the renal medulla may represent a novel mechanism for hypertension in Dahl S rats , and inhibition of Q9GZT9 in the renal medulla could be a therapeutic approach for salt-sensitive hypertension . Involvement of retinoic acid receptor alpha in the stimulation of tissue-type plasminogen-activator gene expression in human endothelial cells . Retinoids stimulate tissue-type plasminogen-activator ( t-PA ) gene expression in human endothelial cells , and are likely to do so by binding to one or more nuclear retinoid receptors . The present study was initiated to identify the retinoid receptor(s) involved in this process . Expression and regulation of retinoic acid receptors ( RARs ) and retinoid X receptors ( RXRs ) were analyzed by Northern-blot analysis of total or poly(A)-rich RNA prepared from cultured human umbilical vein endothelial cells ( HUVEC ) . Prior to any exposure to retinoids , HUVEC express two transcripts for P10276 ( 3.6 kb and 2.8 kb ) , and low levels of transcripts for P10826 ( 3.4 kb and 3.2 kb ) and P13631 ( 3.3 kb and 3.1 kb ) . Two RXR subtypes were identified , RXR-alpha ( 4.8 kb ) and , at a much lower concentration , RXR-beta ( 2.4 kb ) ; no evidence for the presence of RXR-gamma was found . Furthermore , HUVEC express cellular retinol-binding protein I ( P09455 ) and cellular retinoic-acid-binding protein I ( P29762 ) mRNA . Exposure of HUVEC to 1 microM retinoic acid or the DB04942 , Ch55 , led to the induction of the two P10826 mRNAs , RXR-alpha mRNA and P09455 mRNA , whereas the expression of the other receptor and P29762 transcripts did not change appreciably . Using retinoid analogues that bind preferentially to one of the RAR or RXR subtypes , we found evidence that P10276 is involved in the retinoid-induced t-PA expression in HUVEC . This conclusion was strengthened by experiments in which blocking of P10276 with a specific P10276 antagonist , Ro 41-5253 , was demonstrated to suppress the induction of t-PA by retinoids . Interpretation of point-of-care INR results in patients treated with dabigatran . BACKGROUND : Point-of-care devices for measurement of the international normalized ratio ( INR ) are commonly used to monitor therapy and maintain therapeutic levels of anticoagulation in patients treated with vitamin K antagonists . DB06695 , a new oral , reversible direct thrombin inhibitor approved for stroke prevention in patients with atrial fibrillation does not require routine coagulation monitoring . However , case reports have identified falsely elevated point-of-care INR levels in patients treated with dabigatran using one of these devices ( Hemochron ) . This in vitro study was designed to verify this issue . METHODS : We compared INR levels in whole blood and plasma using a Hemochron Jr . Signature+ point-of-care device ( International Technidyne Corporation , Edison , NJ ) with routine laboratory monitoring , using blood from healthy volunteers that was spiked with increasing concentrations of dabigatran . RESULTS : P00734 time and INR levels were increased about 2- to 4-fold with the point-of-care device compared with laboratory measures across the plasma dabigatran concentration range 50-1400 ng/mL . At plasma concentrations of dabigatran likely to be observed in patients , at a dose of 150 mg twice daily ( 60-275 ng/mL ) , whole blood point-of-care INR values increased from 1.7 to 4.0 , versus 1.1 to 1.5 measured with the laboratory coagulometer . Similar differences in prothrombin time were observed in plasma samples . CONCLUSIONS : INR levels in patients taking dabigatran are substantially higher using a Hemochron Jr. point-of-care device compared with laboratory values . We discourage the use of these devices specifically , as well as the use of the INR in general , for measuring the anticoagulant effect of dabigatran .

MH_train_1350

MH_train_1350

MH_train_1350

interacts_with DB00734?

multiple_choice

Augmentation by citalopram of risperidone-induced monoamine release in rat prefrontal cortex . RATIONALE : A typical antipsychotics ( APDs ) , e.g. olanzapine and risperidone , have been reported to be effective adjunctive treatment for depression if selective serotonin ( 5-HT ) reuptake inhibitors ( SSRIs ) alone are ineffective . OBJECTIVES AND METHODS : We utilized microdialysis in awake , freely moving rats to study the effect of risperidone in combination with citalopram , an SSRI , on extracellular 5-HT , dopamine ( DA ) , and norepinephrine ( NE ) efflux in rat medial prefrontal cortex ( mPFC ) . RESULTS : DB00734 ( 1.0 mg/kg , s.c. ) , given alone , significantly increased 5-HT , DA , and NE concentrations in the mPFC . DB00215 ( 10 mg/kg , s.c. ) , by itself , produced a significant increase in 5-HT levels only . The combination of risperidone and citalopram produced significantly greater increases in efflux of both DA and NE than risperidone alone . However , the effect of this combination on extracellular 5-HT concentrations was not significantly different than that of citalopram alone . The augmentation of DA and NE efflux induced by risperidone plus citalopram could be partially blocked by the selective P08908 antagonist , WAY 100635 ( 0.2 mg/kg , s.c. ) . CONCLUSIONS : The results suggest that the ability of atypical APDs to augment the therapeutic efficacy of SSRIs in major depression and treatment-resistant depression may be due , at least in part , to potentiation of SSRI-induced increases in cortical DA and NE . The contributions of P08908 receptor stimulation and 5- Q13049 and alpha2 adrenergic receptor antagonism to this augmentation are discussed . Suppression of tumor growth and metastasis by a P17948 antagonizing peptide identified from a phage display library . Although the P15692 -Flk-1-pathway has been known as the major driving force of angiogenesis , new evidence has shown that P17948 /Flt-1 plays important roles during the neovascularization under pathological conditions including tumor , atherosclerosis and arthritis . In search of Flt-1 receptor antagonizing peptides , we screened a phage display 12-mer-peptide library with recombinant Flt-1 protein . Seven candidate peptides were identified that specifically bound to P15692 receptor Flt-1 , of which peptide F56 ( WHSDMEWWYLLG ) almost abolished P15692 binding to receptor Flt-1 in vitro . In vivo , F56 fused with P00374 ( P00374 -F56 ) inhibited angiogenesis in a P62158 assay . Moreover , P00374 -F56 significantly inhibited the growth of nodules of human gastric cancer cell line MGC-803 in BALB/c nude mice . Histological analyses showed that necrosis of the implanted tumor was markedly enhanced following treatment with P00374 -F56 . In the severe combined immunodeficiency disease ( SCID ) mouse model for studying metastasis of the human breast cancer cell line BICR-H1 , synthetic peptide F56 significantly inhibited tumor growth and lung metastases . Taken together , our results have demonstrated that peptide F56 , as a Flt-1 receptor antagonist , fulfilled the antiangiogenic and antimetastatic effects by specifically interfering with the interaction between P15692 and receptor Flt-1 . Thus , short peptide F56 may have clinical potential in tumor therapy . Effect of N9-methylation and bridge atom variation on the activity of 5-substituted 2,4-diaminopyrrolo[2,3-d]pyrimidines against dihydrofolate reductases from Pneumocystis carinii and Toxoplasma gondii . The effect of N9-methylation and bridge atom variation on inhibitory potency and selectivity of 2,4-diaminopyrrolo[2,3-d]pyrimidines against dihydrofolate reductases ( P00374 ) was studied . Specifically three nonclassical 2,4-diamino-5-((N-methylanilino)methyl)pyrrolo[2,3-d]pyrimidines with 2',5'-dimethoxyphenyl ( 2 ) , 3',4'-dichlorophenyl ( 3 ) , 1'-naphthyl ( 4 ) , one classical analogue with a 4'-L-glutamate substituent ( 10 ) , and four nonclassical 2,4-diamino-5-((phenylthio)methyl)pyrrolo[2,3-d]pyrimidines with 3',4'-dimethoxyphenyl ( 5 ) , 3',4'-dichlorophenyl ( 6 ) , 1'-naphthyl ( 7 ) , and 2'-naphthyl ( 8 ) substituents were synthesized . The classical and nonclassical analogues were obtained by displacement of the intermediate 2,4-diamino-5-bromomethylpyrrolo[2,3-d]pyrimidine , 14 , with appropriately substituted N-methylaniline , thiophenols , or 4-(N-methylamino)benzoyl-L-glutamate . Compounds 2-8 and 10 were evaluated against Pneumocystis carinii ( pc ) , Toxoplasma gondii ( tg ) , and rat liver ( rl ) DHFRs . The N-methyl and thiomethyl analogues were more inhibitory than their corresponding anilinomethyl analogues ( previously reported ) against all three DHFRs . The inhibitory potency of these analogues was greater against rlDHFR than against tgDHFR which resulted in a loss of selectivity for tgDHFR compared to the N9-H analogues . The classical N9-methyl analogue 10 was more potent and about 2-fold more selective against tgDHFR than its corresponding desmethyl analogue . All of the analogues , 2-8 and 10 , were more selective than trimetrexate ( DB01157 ) against pcDHFR ( except 4 ) and significantly more selective than DB01157 against tgDHFR . Lowering of body core temperature by exposure to a cold environment and by a P08908 agonist : effects on physiological and psychological variables and blood serotonin levels . The present study was designed to compare the effects of a pharmacologically induced decrease in body core temperature to the effects observed with lowering of body temperature by exposure to a cold environment . Our special interest was the involvement of 5-HT in thermoregulatory responses . Sixty healthy male volunteers were randomly assigned to one of the following conditions : exposure to normal ambient temperature ( 28 degrees C ) and placebo , exposure to cold ambient temperature ( 5 degrees C ) and placebo , or normal ambient temperature and 10 mg of the partial P08908 agonist ipsapirone . As indicators of physiological responses to lowering of body temperature , tympanic temperature , skin temperature , P15941 , metabolic rate , and heart rate were monitored and saliva cortisol levels and peripheral 5-HT concentrations were determined . In addition , ratings on ambient temperature , thermal discomfort , and feelings of irritability were obtained . While lowering of body core temperature was associated with marked counterregulations ( decrease of skin temperature , increase in P15941 and metabolic rate ) and feelings of discomfort , this was not observed with ipsapirone . An increase in cortisol levels was primarily observed in the ipsapirone group and was not reflected by respective changes in whole blood or platelet 5-HT indicating that brain and platelet 5-HT are not related . Protective effects of mineralocorticoid receptor blockade against neuropathy in experimental diabetic rats . AIMS : P08235 ( MR ) blockade is an effective treatment for hypertension and diabetic nephropathy . There are no data on the effects of MR blockade on diabetic peripheral neuropathy ( DPN ) . The aim of this study was to determine whether MRs are present in the peripheral nerves and to investigate the effectiveness of MR blockade on DPN in streptozotocin ( Q11206 ) -induced diabetic rats . METHODS : Expression of MR protein and messenger RNA ( mRNA ) was examined in the peripheral nerves using Western blot analysis and RT-PCR . We next studied the effects of the selective MR antagonist eplerenone and the angiotensin II receptor blocker candesartan on motor and sensory nerve conduction velocity ( NCV ) , morphometric changes and cyclooxygenase-2 ( P35354 ) gene and NF-κB protein expression in the peripheral nerves of Q11206 -induced diabetic rats . RESULTS : Expression of MR protein and mRNA in peripheral nerves was equal to that in the kidney . Motor NCV was significantly improved by 8 weeks of treatment with either eplerenone ( Q04695 ± 1.2 m/s ) or candesartan ( 46.4 ± 6.8 m/s ) compared with control diabetic rats ( 33.7 ± 2.0 m/s ) ( p < 0.05 ) . Sensory NCV was also improved by treatment with candesartan or eplerenone in diabetic rats . DB00700 and candesartan caused significant improvement in mean myelin fibre area and mean myelin area compared with control diabetic rats ( p < 0.05 ) . P35354 mRNA and NF-κB protein were significantly elevated in the peripheral nerves of diabetic rats compared with control rats , and treatment with eplerenone or candesartan reduced these changes in gene expression ( p < 0.05 ) . CONCLUSION : MR blockade may have neuroprotective effects on DPN . Rapid T cell repopulation after rabbit anti-thymocyte globulin ( DB00098 ) treatment is driven mainly by cytomegalovirus . Rabbit anti-thymocyte globulin ( DB00098 ) induces a long-lasting lymphocytopenia . P01730 (+) T cells remain depleted for up to 2 years , whereas the CD8(+) T cell compartment is refilled rapidly by highly differentiated P26842 (-) CD45RA(+) CD57(+) effector-type cells . Because the presence of these highly differentiated CD8(+) T cells has been associated with cytomegalovirus ( CMV ) infection , we questioned to what extent restoration of CMV T cell immunity contributes to the re-emergence of T cells following DB00098 treatment . We compared T cell repopulation in six CMV-seropositive patients with CMV reactivation ( reactivating CMV(+) ) to that in three CMV(+) patients without reactivation ( non-reactivating CMV(+) ) , and to that in three CMV-seronegative recipients receiving a kidney from a CMV-seronegative donor ( CMV(-/-) ) . All patients received DB00098 because of acute allograft rejection . Total P01730 and CD8 counts , frequency and phenotype of virus-specific CD8(+) T cells were determined . In reactivating CMV(+) patients , total CD8(+) T cells reappeared rapidly , whereas in non-reactivating CMV(+) patients they lagged behind . In CMV(-/-) patients , CD8(+) T cell counts had not yet reached pretransplant levels after 2 years . CMV reactivation was indeed followed by a progressive accumulation of CMV-specific CD8(+) T cells . During lymphocytopenia following DB00098 treatment , serum interleukin ( IL ) -7 levels were elevated . Although this was most prominent in the CMV-seronegative patients , it did not result in an advantage in T cell repopulation in these patients . Repopulated CD8(+) T cells showed increased skewing in their Vβ repertoire in both CMV(-/-) and reactivating CMV-seropositive patients . We conclude that rapid T cell repopulation following DB00098 treatment is driven mainly by CMV . Filamin A in somatostatin and dopamine receptor regulation in pituitary and the role of DB02527 /PKA dependent phosphorylation . Molecular mechanisms underlying resistance of pituitary tumors to somatostatin ( SS ) and dopamine ( DA ) analogues treatment are not completely understood . Resistance has been associated with defective expression of functional somatostatin and dopamine receptors P30874 , P35346 , and P14416 , respectively . Recently , a role of cytoskeleton protein filamin A ( P21333 ) in P14416 and SSTR receptors expression and signaling in PRL- and GH-secreting tumors , respectively , has been demonstrated , first revealing a link between P21333 expression and responsiveness of pituitary tumors to pharmacological therapy . No molecular events underlying the reduction of P21333 levels in resistant tumors have been so far identified . P21333 can be phosphorylated by PKA on Ser2152 , with increased P21333 resistance to cleavage by calpain and conformational changes affecting P21333 regions involved in P30874 and P14416 binding and signal transduction . In this respect , the effect of DB02527 /PKA pathway in the regulation of P21333 stability and/or function by modulating its phosphorylation status could assume particular importance in pituitary , where DB02527 cascade plays a crucial role in pituitary cell functions and tumorigenesis . This review will discuss the role of P21333 in the regulation of the main GPCRs target of pharmacological treatment of pituitary tumors , that is , P30874 and P14416 , focusing on the effects of DB02527 /PKA-mediated P21333 phosphorylation on P21333 biological functions . Effects of serotonin on expression of the P01130 family member Q92673 and 7-ketocholesterol-induced apoptosis in human vascular smooth muscle cells . Serotonin ( 5-HT ) is a known mitogen for vascular smooth muscle cells ( VSMCs ) . The dedifferentiation and proliferation/apoptosis of VSMCs in the arterial intima represent one of the atherosclerotic changes . Q92673 , a member of low-density lipoprotein receptor family , may contribute to the proliferation of VSMCs in neointimal hyperplasia . We conducted an in vitro study to investigate whether 5-HT is involved in Q92673 expression in human VSMCs and apoptosis of VSMCs induced by 7-ketocholesterol ( 7KCHO ) , an oxysterol that destabilizes plaque . 5-HT enhanced the proliferation of VSMCs , and this effect was abolished by sarpogrelate , a selective 5- Q13049 receptor antagonist . Sarpogrelate also inhibited the 5-HT-enhanced Q92673 mRNA expression in VSMCs . Furthermore , 5-HT suppressed the 7KCHO-induced apoptosis of VSMCs via caspase-3/7-dependent pathway . These findings provide new insights on the changes in the differentiation stage of VSMCs mediated by 5-HT . DB06016 , an orally active D2/D3 receptor antagonist , for the potential treatment of schizophrenia , bipolar mania and depression . DB06016 ( RGH-188 ) , which is being codeveloped by Gedeon Richter Ltd , Forest Laboratories Inc and Mitsubishi Tanabe Pharma Corp , is a novel putative antipsychotic drug that exerts partial agonism at dopamine D2/D3 receptors , with preferential binding to D3 receptors , and partial agonism at serotonin P08908 receptors . Its activity at D2/D3 receptors may be lower than that of the prototype partial agonist aripiprazole . The antipsychotic activity of cariprazine was demonstrated in animal models , and data also suggest that the propensity for extrapyramidal side effects is low and that the drug may have procognitive properties . DB06016 is rapidly absorbed , with high oral bioavailability and a long plasma elimination t1/2 . DB06016 is in phase III clinical trials in patients with schizophrenia and in patients with bipolar disorder . Data from phase II trials in patients with schizophrenia and bipolar mania indicate that the drug has antipsychotic and antimanic properties that are superior to placebo . With its unique receptor affinity profile , cariprazine may represent a potential enrichment of the therapeutic armamentarium for schizophrenia and affective disorders . Its activity against the cognitive deficits associated with schizophrenia has to be carefully investigated . A randomized , placebo-controlled study of the effects of the p38 MAPK inhibitor SB- DB05250 on blood biomarkers of inflammation in P48444 patients . The p38 mitogen-activated protein kinase ( MAPK ) signaling upregulates inflammation and is known to be increased in chronic obstructive pulmonary disease ( P48444 ) . The authors assessed the pharmacology of the novel p38 MAPK inhibitor SB- DB05250 using blood biomarkers in P48444 . Seventeen P48444 patients ( forced expiratory volume in 1 second 50 % -80 % predicted ) using short-acting bronchodilators participated in a double-blind , double-dummy , randomized , crossover study . Patients received single oral doses of SB- DB05250 7.5 mg and 25 mg , prednisolone 10 mg and 30 mg , and placebo . Blood was obtained predose and at 1 , 2 , 6 , and 24 hours postdose . Whole-blood sorbitol-induced phosphorylated ( p ) heat shock protein ( HSP ) 27 levels as a marker of p38 pathway activation and lipopolysaccharide-induced tumor necrosis factor ( P01375 ) -alpha production were assessed . Both doses of SB- DB05250 , but not prednisolone , significantly ( P < .0001 ) reduced weighted mean ( WM ) pHSP27 ( 0-6 hours ) by 58 % compared with placebo . WM P01375 production ( 0-24 hours ) was significantly reduced compared with placebo by SB- DB05250 25 mg ( 40 % , P = .005 ) and 7.5 mg ( 33.4 % , P = .02 ) , while prednisolone 30 mg and 10 mg caused 81.5 % and 58.2 % suppression , respectively ( both P < .0001 ) . SB- DB05250 inhibited the p38 MAPK pathway to a greater degree than prednisolone did . SB- DB05250 inhibited P01375 production . SB- DB05250 is a potent p38 MAPK inhibitor that potentially suppresses inflammation in P48444 . Q07973 as a potential target for cancer therapy . Increasing evidence has accumulated to suggest that vitamin D may reduce the risk of cancer through its biologically active metabolite , DB00136 , which inhibits proliferation and angiogenesis , induces differentiation and apoptosis , and regulates many other cellular functions . Thus , it is plausible to assume that rapid clearance of DB00136 by highly expressed Q07973 could interrupt the normal physiology of cells and might be one cause of cancer initiation and progression . In fact , enhancement of Q07973 expression has been reported in literature for many cancers . Based on these findings , Q07973 -specific inhibitors and vitamin D analogs which are resistant to Q07973 -dependent catabolism might be useful for cancer treatment . Q07973 -specific inhibitor VID400 , which is an azole compound , markedly enhanced and prolonged the antiproliferative activity of DB00136 in the human keratinocytes . Likewise , Q07973 -resistant analogs such as 2α-(3-hydroxypropoxy)- DB00136 ( O2C3 ) and its P06681 -epimer ED-71 ( DB05295 ) , and 19nor- 2α-(3-hydroxypropyl)- DB00136 ( MART-10 ) showed potent biological effects . Our in vivo studies using rats revealed that MART-10 had a low calcemic effect , which is a suitable property as an anticancer drug . Much lower affinity of MART-10 for vitamin D binding protein ( DBP ) as compared with DB00136 may be related to its more potent cellular activities . Based on these results , we conclude that ( 1 ) high affinity for P11473 , ( 2 ) resistance to Q07973 -dependent catabolism , ( 3 ) low affinity for DBP , and ( 4 ) low calcemic effect may be required for designing potent vitamin D analogs for cancer treatment . Neurological impairment in experimental antiphospholipid syndrome is associated with increased ligand binding to hippocampal and cortical serotonergic P08908 receptors . The antiphospholipid syndrome ( APS ) is an autoimmune disease where the presence of high titers of circulating autoantibodies causes thrombosis with consecutive infarcts . In experimental APS ( eAPS ) , a mouse model of APS , behavioral abnormalities develop in the absence of vessel occlusion or infarcts . Using brain hemispheres of control and eAPS mice with documented neurological and cognitive deficits , we checked for lymphocytic infiltration , activation of glia and macrophages , as well as alterations of ligand binding densities of various neurotransmitter receptors to unravel the molecular basis of this abnormal behavior . Lymphocytic infiltrates were immunohistochemically characterized using antibodies against CD3 , P01730 , CD8 and forkhead box P09131 ( Foxp3 ) , respectively . P14136 , Iba1 and P34810 -immunohistochemistry was performed , to check for activation of astrocytes , microglia and macrophages . Ligand binding densities of DB01221 , AMPA , GABAA and P08908 receptors were analyzed by in vitro receptor autoradiography . No significant inflammatory reaction occurred in eAPS mice . There was neither activation of astrocytes or microglia nor accumulation of macrophages . Binding values of excitatory and inhibitory neurotransmitter receptors were largely unchanged . However , ligand binding densities of the modulatory serotonergic P08908 receptors in the hippocampus and in the primary somatosensory cortex of eAPS mice were significantly upregulated which is suggested to induce the behavioral abnormalities observed . DB08875 ( DB05153 ) for the treatment of locally advanced or metastatic progressive medullary thyroid cancer . DB08875 ( DB05153 ) is an oral multiple receptor tyrosine kinase inhibitor manufactured by Exelixis Inc. , CA , USA . It mainly inhibits three tyrosine kinase receptors : MET , P35968 and P07949 . In both preclinical and clinical studies it has been shown to inhibit tumor angiogenesis , invasiveness and metastases . The most frequent side effects are fatigue , diarrhea , decreased appetite , nausea , weight loss and palmar-plantar erythrodysesthesia . A Phase III clinical trial ( EXAM study ) of DB05153 versus placebo in advanced and progressive medullary thyroid cancer showed a 28 versus 0 % overall response rate and a progression-free survival of 11.2 versus 4.0 months ( hazard ratio : 0.28 ; 95 % CI : 0.19-0.40 ; p < 0.0001 ) in patients treated with cabozantinib and placebo , respectively . The drug has been approved by the US FDA for the treatment of advanced/progressive metastatic medullary thyroid cancer in the USA . The P15941 is now evaluating its approval in Europe . DB09302 inhibits atherosclerosis , improves the plaque morphology , and enhances the effects of a statin . Q8NBP7 ( Q8NBP7 ) inhibition is a potential novel strategy for treatment of CVD . DB09302 is a fully human Q8NBP7 monoclonal antibody in phase 3 clinical development . We evaluated the antiatherogenic potential of alirocumab in P02649 *3Leiden. P11597 mice . Mice received a Western-type diet and were treated with alirocumab ( 3 or 10 mg/kg , weekly subcutaneous dosing ) alone and in combination with atorvastatin ( 3.6 mg/kg/d ) for 18 weeks . DB09302 alone dose-dependently decreased total cholesterol ( -37 % ; -46 % , P < 0.001 ) and TGs ( -36 % ; -39 % , P < 0.001 ) and further decreased cholesterol in combination with atorvastatin ( -48 % ; -58 % , P < 0.001 ) . DB09302 increased hepatic P01130 protein levels but did not affect hepatic cholesterol and TG content . Fecal output of bile acids and neutral sterols was not changed . DB09302 dose-dependently decreased atherosclerotic lesion size ( -71 % ; -88 % , P < 0.001 ) and severity and enhanced these effects when added to atorvastatin ( -89 % ; -98 % , P < 0.001 ) . DB09302 reduced monocyte recruitment and improved the lesion composition by increasing the smooth muscle cell and collagen content and decreasing the macrophage and necrotic core content . DB09302 dose-dependently decreases plasma lipids and , as a result , atherosclerosis development , and it enhances the beneficial effects of atorvastatin in P02649 *3Leiden. P11597 mice . In addition , alirocumab improves plaque morphology . DB04894 labeled with Tc-99m for imaging tumors . DB04894 ( RC-160 ) , an octapeptide analog of somatostatin , has a high affinity for somatostatin receptor subtypes P30874 and P35346 . DB04894 binds differently to the tumors of the breast , ovary , exocrine pancreas , prostate and colon , than octreotide another octapeptide analog of somatostatin . DB04894 was labeled with Tc-99m , a radionuclide highly suitable for scintigraphic imaging . The labeling procedure was simple , produced > 70 % yields and could be applicable to label other peptides containing a cystine bridge . HPLC analysis showed that the tracer was stable when Tc-99m-RC-160 was challenged with 100 fold molar excess DTPA ( diethylenetriaminepentaacetic acid ) , HSA ( human serum albumin ) or cysteine and incubated at 37 degrees C for 4 h . HPLC analysis of urine samples obtained from mice that received Tc-99m-RC-160 showed that the preparation was stable in vivo . Rat brain cortex membrane receptor displacement assays showed that the Kd values for Tc-99m-RC-160 ( 71x10(-9) M ) and Tc-99m-octreotide ( 86x10(-9) M ) ( Sandostatin(R) ) were in nM range , and were similar to that for I-125-RC-160 ( 46x10(-9) M ) . High binding affinity of Tc-99m-RC-160 for human breast tumor cells SKBR-3 was also observed . These results suggest that Tc-99m-RC-160 is worthy of evaluation as an agent for scintigraphic imaging of tumors rich in somatostatin receptor subtypes P30874 and P35346 . Signaling pathways mediating induction of the early response genes prostaglandin G/H synthase-2 and egr-1 by serotonin via 5- Q13049 receptors . Signaling pathways responsible for serotonin ( 5-HT ) -mediated induction of early response genes prostaglandin G/H synthase-2 ( P35354 , cyclooxygenase-2 ) and egr-1 were investigated in rat mesangial cells . Gene induction by 5-HT was dependent on 5- Q13049 receptors that were pertussis toxin insensitive indicating coupling to a G-protein of the Gq family . Binding of 5-HT to this receptor activates phosphatidylinositol-specific phospholipase C ( P98160 ) and release of Ca2+ from internal stores , but this activation was not related to P35354 mRNA expression . Similarly , P19957 kinase was not involved in 5-HT signaling . Instead , inhibition of phosphatidylcholine-specific P98160 interfered with P35354 and egr-1 mRNA induction , suggesting this enzyme as a link between 5- Q13049 receptors and protein kinase C , an essential part of 5-HT-mediated signaling . The Q96HU1 kinase pathway was identified as common signaling pathway of 5-HT or phorbol ester-induced gene expression . Increase of intracellular DB02527 by forskolin or dibutyryl DB02527 did not induce P35354 or egr-1 mRNA expression by itself , but strongly inhibited 5-HT-mediated mRNA induction . P35354 mRNA and protein induction by 5-HT was also abolished by chelation of Ca2+ ions by EGTA , suggesting involvement of Ca2+-dependent enzymes . In contrast , egr-1 mRNA expression was superinduced in the presence of EGTA . Induction of Egr-1 protein was not changed by EGTA hinting to Ca2+-sensitive posttranscriptional steps . Activation of the Gq-coupled 5- Q13049 receptor thus leads to the expression of the early response genes P35354 and egr-1 , using common as well as differing signaling elements that allow differential regulation of the expression of these genes that are functionally related to renal hemodynamics and proliferation of mesangial cells , respectively . P14416 occupancy by risperidone : implications for the timing and magnitude of clinical response . The objective of the study is to investigate whether dopamine D2 receptor occupancy by risperidone and plasma levels over time can account for therapeutic efficacy and the latency period to response . Thirty-eight examinations with (123)I-IBZM single photon emission computed tomography were performed on 22 patients with schizophrenia , at diagnosis , 48 h after starting risperidone treatment and at a stable dose . DB00734 plasma levels were determined and psychopathologic evaluations ( Brief Psychiatric Rating Scale , Positive and Negative Syndrome Scale ) were carried out . No differences in the striatal/occipital ( S/O ) ratio or plasma levels were found between examinations at the 48-h time point and when a stable dose level had been established , so these parameters could not account for the latency period required for clinical response . D2 receptor occupancy at 48 h correlated positively with clinical improvement after 2 weeks of treatment . Therefore , if these results are confirmed , D2 receptor occupancy at the beginning of treatment with risperidone may be a predictor of subsequent clinical response . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . Differential regulation of the serotonin 1 A transcriptional modulators five prime repressor element under dual repression-1 and nuclear-deformed epidermal autoregulatory factor by chronic stress . Chronic stress is known to affect brain areas involved in learning and emotional responses . These changes , thought to be related to the development of cognitive deficits are evident in major depressive disorder and other stress-related pathophysiologies . The serotonin-related transcription factors ( Q6P1N0 / Q6P1N0 ; five prime repressor element under dual repression/coiled-coil P06681 domain 1a , and O75398 /Deaf-1 ; nuclear-deformed epidermal autoregulatory factor ) are two important regulators of the P08908 receptor . Using Western blotting and quantitative real-time polymerase chain reaction ( qPCR ) we examined the expression of mRNA and proteins for Q6P1N0 , O75398 , and the P08908 receptor in the prefrontal cortex ( P27918 ) of male rats exposed to chronic restraint stress ( CRS ; 6 h/day for 21 days ) . After 21 days of CRS , significant reductions in both Q6P1N0 mRNA and protein were observed in the P27918 ( 36.8 % and 32 % , respectively ; P < 0.001 ) , while the levels of both O75398 protein and mRNA did not change significantly . Consistent with reduced Q6P1N0 protein , P08908 receptor mRNA levels were equally upregulated in the P27918 , while protein levels actually declined , suggesting post-transcriptional receptor downregulation . The data suggest that CRS produces distinct alterations in the serotonin system specifically altering Q6P1N0 and the P08908 receptor in the P27918 of the male rat while having no effect on O75398 . These results point to the importance of understanding the mechanism for the differential regulation of Q6P1N0 and O75398 in the P27918 as a basis for understanding the related effects of chronic stress on the serotonin system ( serotonin-related transcription factors ) and stress-related disorders like depression .

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interacts_with DB08815?

multiple_choice

Preclinical evaluation of a novel water-soluble chlorin E6 derivative ( O43927 1010 ) as photosensitizer for the closure of the neovessels . In the present study , photodynamic activity of a novel photosensitizer ( PS ) , Chlorin e(6)-2.5 N-methyl-d-glucamine ( O43927 1010 ) , was evaluated using the chorioallantoic membrane ( P62158 ) as an in vivo model . After intravenous ( i.v. ) injection of O43927 1010 into the P62158 vasculature , the applicability of this drug for photodynamic therapy ( PDT ) was assessed in terms of fluorescence pharmacokinetics , i.e. leakage from the P62158 vessels , and photothrombic activity . The influence of different PDT parameters including drug and light doses on the photodynamic activity of O43927 1010 has been investigated . It was found that , irrespective of drug dose , an identical continuous decrease in fluorescence contrast between the drug inside and outside the blood vessels was observed . The optimal treatment conditions leading to desired vascular damage were obtained by varying drug and light doses . Indeed , observable damage was achieved when irradiation was performed at light doses up to 5 J/cm(2) 1 min after i.v. injection of drug doses up to 0.5 mg/kg body weight(b.w.) . However , when irradiation with light doses of more than 10 J/cm(2) was performed 1 min after injection of drug doses up to 2 mg/kg body weight , this led to occlusion of large blood vessels . It has been demonstrated that it is possible to obtain the desired vascular occlusion and stasis with O43927 1010 for different combinations of drug and/or light doses . Whole blood lead concentration and erythrocyte delta-aminolevulinic acid dehydratase ( P13716 ) activity in selected canine populations in Greece . In a total number of 275 dogs of various ages , sex and breed , blood lead concentrations ( O43927 ) and erythrocyte P13716 activity were measured . Sixty-six of the dogs were living in lead mining areas ( Group A ) , 157 in urban areas ( Group B ) and 52 in rural areas ( Group C ) of Greece . Mean O43927 differed significantly ( P < 0.05 ) between locations and were 326,97 and 68 micrograms/L , respectively . Mean P13716 activity was significantly different ( P < 0.05 ) only between Groups A and B as between groups A and C . A significant ( P < 0.05 ) negative correlation existed between O43927 and P13716 activity . A normal range of erythrocyte P13716 activity of 807-992 mumol/ DB02272 /LRBC/h was established for dogs . None of the 33 Group A dogs and 2 of the Group B dogs that had a O43927 of 350 micrograms/L presented clinical signs indicating acute or chronic lead intoxication . No erythrocyte basophilic stippling or large number of nucleated red blood cells were seen in the 30 dogs of Group A with O43927 > 350 micrograms/L . The PEPvIII-KLH ( DB05374 ) vaccine in glioblastoma multiforme patients . Conventional therapies for glioblastoma multiforme ( GBM ) fail to target tumor cells exclusively , resulting in non-specific toxicity . Immune targeting of tumor-specific mutations may allow for more precise eradication of neoplastic cells . P00533 variant III ( EGFRvIII ) is a tumor-specific mutation that is widely expressed in GBM and other neoplasms and its expression enhances tumorigenicity . This in-frame deletion mutation splits a codon , resulting in a novel glycine at the fusion junction producing a tumor-specific epitope target for cellular or humoral immunotherapy . We have previously shown that vaccination with a peptide that spans the EGFRvIII fusion junction ( PEPvIII-KLH/ DB05374 ) is an efficacious immunotherapy in syngeneic murine models . In this review , we summarize our results in GBM patients targeting this mutation in multiple , multi-institutional Phase II immunotherapy trials . These trials demonstrated that a selected population of GBM patients who received vaccines targeting EGFRvIII had an unexpectedly long survival time . Further therapeutic strategies and potential pitfalls of using this approach are discussed . Pharmacological Q9BWK5 mapping of age-associated changes in basal ganglia circuitry of awake rhesus monkeys . While the pathophysiological changes induced by the loss of dopamine innervation in the basal ganglia by Parkinson 's disease ( PD ) are well studied , little is known about functional changes in the neural circuitry of this area during normal aging . Here we report the first survey of age-associated changes in the basal ganglia of behaviorally characterized , awake rhesus monkeys , using pharmacological Q9BWK5 to map responses to dopaminergic stimulation . DB00714 , a mixed D(1)/ P14416 agonist , evoked little change in the substantia nigra ( SN ) of aged animals while significantly reducing activation in young adult monkeys . Compared to young animals , both apomorphine and DB01576 ( which increases synaptic dopamine levels ) significantly increased activation of the aged rhesus globus pallidus externa ( GPe ) . In addition , the aged animals showed decreased activity in the putamen in response to DB01576 administration . Although the responses in the SN and putamen of the aged monkeys differed from those in animal models of PD , the apomorphine-evoked activation of their GPe corresponded with apomorphine-induced increases in neuronal activity seen in Parkinson 's patients and animal models . Given the major role of the GPe in regulating motor behavior , the altered responses in the aged GPe may contribute significantly to the motor slowing and movement dysfunctions characterizing advanced age . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . P55157 inhibitor decreases plasma cholesterol levels in P01130 -deficient WHHL rabbits by lowering the VLDL secretion . To examine whether a microsomal triglyceride transfer protein ( P55157 ) -inhibitor is effective in patients with homozygous familial hypercholesterolemia , we administered ( 2S ) -2-cyclopentyl-2-[4-[(2,4-dimethyl-9H-pyrido[2,3-b]indol-9-yl)methyl]phenyl]-N- [ ( 1S ) -2-hydroxy-1-phenylethyl ] ethanamide ( DB04852 ) , a new P55157 inhibitor , to low-density lipoprotein ( LDL ) -receptor-deficient Watanabe heritable hyperlipidemic ( WHHL ) rabbits at doses of 3 , 6 , and 12 mg/kg for 4 weeks . In the 12 mg/kg group , the plasma cholesterol and triglyceride levels were decreased by 70 % and 45 % , respectively , and the very low-density lipoprotein ( VLDL ) secretion rate was decreased by 80 % . The composition of newly secreted VLDL was similar in each group . This suggests that DB04852 diminished the number of VLDL particles secreted from the liver . Although the ratio of vitamin E/LDL was not altered by DB04852 , triglyceride accumulation and a decrease in vitamin E were observed in the liver . In conclusion , an inhibition of VLDL secretion led to a decrease of plasma LDL in WHHL rabbits , and P55157 inhibitors should have hypolipidemic effects against homozygous familial hypercholesterolemia . Poly(ADP-ribose) metabolism in X-irradiated Chinese hamster cells : its relation to repair of potentially lethal damage . DB02701 -adenine dinucleotide ( NAD+ ) is the substrate used by cells in poly(ADP-ribose) synthesis . X-irradiation of log-phase Chinese hamster cells caused a rapid decrease in NAD+ levels which was linearly dependent on radiation dose . The activity of ADP-ribosyl transferase ( P09874 ) also increased linearly with radiation dose . The decrease of NAD+ was slower , and the increase in P09874 activity was less pronounced , in a radiation sensitive line , V79- AL162 /S-10 . An inhibitor of P09874 , m-aminobenzamide , largely prevented the depletion of cellular NAD+ and reduced the rate at which P09874 activity disappeared during post-irradiation incubation . Post-irradiation treatment with hypertonic buffer or with medium containing D2O -- which inhibit repair of radiation-induced potentially lethal damage -- enhanced the depletion of NAD+ and prevented the reduction in P09874 activity following irradiation . The characteristics of the effects of treatment with hypertonic buffer on NAD+ metabolism were qualitatively similar to the effects that such treatment has on radiation-induced cell killing . These results suggest that poly(ADP-ribose) synthesis after irradiation plays a role in the repair of potentially lethal damage . DB02426 effects on brown-fat mitochondria imply that the adenine nucleotide translocator isoforms P12235 and P05141 may be responsible for basal and fatty-acid-induced uncoupling respectively . In brown-fat mitochondria , fatty acids induce thermogenic uncoupling through activation of P25874 ( uncoupling protein 1 ) . However , even in brown-fat mitochondria from P25874 -/- mice , fatty-acid-induced uncoupling exists . In the present investigation , we used the inhibitor CAtr ( carboxyatractyloside ) to examine the involvement of the ANT ( adenine nucleotide translocator ) in the mediation of this P25874 -independent fatty-acid-induced uncoupling in brown-fat mitochondria . We found that the contribution of ANT to fatty-acid-induced uncoupling in P25874 -/- brown-fat mitochondria was minimal ( whereas it was responsible for nearly half the fatty-acid-induced uncoupling in liver mitochondria ) . As compared with liver mitochondria , brown-fat mitochondria exhibit a relatively high ( P25874 -independent ) basal respiration ( ' proton leak ' ) . Unexpectedly , a large fraction of this high basal respiration was sensitive to CAtr , whereas in liver mitochondria , basal respiration was CAtr-insensitive . Total ANT protein levels were similar in brown-fat mitochondria from wild-type mice and in liver mitochondria , but the level was increased in brown-fat mitochondria from P25874 -/- mice . However , in liver , only Ant2 mRNA was found , whereas in brown adipose tissue , Ant1 and Ant2 mRNA levels were equal . The data are therefore compatible with a tentative model in which the P05141 isoform mediates fatty-acid-induced uncoupling , whereas the P12235 isoform may mediate a significant part of the high basal proton leak in brown-fat mitochondria . The effectiveness of lurasidone as an adjunct to lithium or divalproex in the treatment of bipolar disorder . The majority of patients with bipolar disorder spend a lot of time in depressive episodes that impose a great burden on patients , caregivers , and society and accounts for the largest part of the morbidity-mortality of the illness . DB08815 is an atypical antipsychotic with a potent binding affinity as antagonist for D2 , 5- Q13049 , P34969 , and partial agonist at P08908 receptors . Affinity for other receptors as H1 and muscarinic were negligible . DB08815 was approved in 2010 for the treatment of schizophrenia and recently , 2013 , for bipolar depression in monotherapy and an adjunct to lithium or valproate . Clinical trials have established that lurasidone adjuvant to lithium or valproate has more efficacy than the placebo and is associated with minimal weight gain and no clinically meaningful alterations in glucose , lipids , or the QT interval . Additional studies are desirable to know the clinical profile of lurasidone in long-term treatment , in patients with bipolar II disorders , and versus other antipsychotic agents . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Effects of serotonin on expression of the P01130 family member Q92673 and 7-ketocholesterol-induced apoptosis in human vascular smooth muscle cells . Serotonin ( 5-HT ) is a known mitogen for vascular smooth muscle cells ( VSMCs ) . The dedifferentiation and proliferation/apoptosis of VSMCs in the arterial intima represent one of the atherosclerotic changes . Q92673 , a member of low-density lipoprotein receptor family , may contribute to the proliferation of VSMCs in neointimal hyperplasia . We conducted an in vitro study to investigate whether 5-HT is involved in Q92673 expression in human VSMCs and apoptosis of VSMCs induced by 7-ketocholesterol ( 7KCHO ) , an oxysterol that destabilizes plaque . 5-HT enhanced the proliferation of VSMCs , and this effect was abolished by sarpogrelate , a selective 5- Q13049 receptor antagonist . Sarpogrelate also inhibited the 5-HT-enhanced Q92673 mRNA expression in VSMCs . Furthermore , 5-HT suppressed the 7KCHO-induced apoptosis of VSMCs via caspase-3/7-dependent pathway . These findings provide new insights on the changes in the differentiation stage of VSMCs mediated by 5-HT . DB05229 sodium , prostacyclin analogue , attenuates glomerular hyperfiltration and glomerular macrophage infiltration by modulating ecNOS expression in diabetic rats . Stable prostacyclin analogue , beraprost sodium ( BPS ) has recently been reported to attenuate glomerular hyperfiltration in diabetic rats , however , the mechanism has been still unknown . We previously reported that overexpression of endothelial cell nitric oxide synthase ( ecNOS ) in afferent arterioles and glomeruli induce inappropriate dilatation of afferent arterioles and glomerular hyperfiltration through overproduction of nitric oxide in early stage of diabetic nephropathy . In this study , we tested the hypothesis that BPS ameliorates glomerular hyperfiltration through modulating ecNOS expression in diabetic nephropathy . Furthermore , we examined the effects of BPS on the expression of intercellular adhesion molecule-1 ( P05362 ) and macrophage infiltration in diabetic glomeruli , because glomerular hyperfiltration induces the expression of P05362 resulting in macrophage infiltration . Male Sprague-Dawley ( SD ) rats were administered continuously with BPS for 4 weeks after induction of diabetes by streptozotocin . In diabetic rats , the diameters of afferent arterioles , glomerular volume , creatinine clearance and urinary excretion of albumin and NO2/NO3 were increased as compared with non-diabetic control rats . Treatment with BPS improved these changes . The expression of ecNOS was increased in afferent arterioles and glomeruli in diabetic rats and suppressed by BPS . P43119 was expressed along afferent arterioles . Our results suggest that BPS attenuates glomerular hyperfiltration by modulating ecNOS expression in early stage of diabetic nephropathy . Moreover , BPS may inhibit P05362 -dependent infiltration of macrophages in diabetic glomeruli . P43119 -induced P40763 phosphorylation in human erythroleukemia cells is mediated via Galpha(s) and Galpha(16) hybrid signaling . Human prostacyclin receptor ( hIP ) stimulates P40763 via pertussis toxin-insensitive G proteins in human erythroleukemia ( HEL ) cells . Since hIP can utilize G(s) and G(q) proteins for signal transduction and that both G proteins can induce P40763 phosphorylation and activation via complex signaling networks , we sought to determine if one of them is predominant in mediating the hIP signal . Stimulation of P40763 DB00135 (705) and DB00133 (727) phosphorylations by the IP-specific agonist , cicaprost , was sensitive to inhibition of protein kinase A , phospholipase Cbeta , protein kinase C , calmodulin-dependent protein kinase II and O60674 /3 . Unlike Galpha(16)-mediated regulation of P40763 in the same cells , cicaprost-induced P40763 DB00135 (705) phosphorylation was resistant to inhibition of Src and MEK while P40763 DB00133 (727) phosphorylation distinctly required phosphatidylinositol-3 kinase . This unique inhibitor-sensitivity pattern of P40763 phosphorylation was reproduced in HEL cells by stimulating the G(16)-coupled C5a receptor in the presence of dibutyryl- DB02527 , suggesting that the change in inhibitor-sensitivity was due to activation of the G(s) pathway . This postulation was confirmed by expressing constitutively active Galpha(16)QL and Galpha(s)QL in human embryonic kidney 293 cells and the inhibitor-sensitivity of Galpha(16)QL-induced P40763 phosphorylations could be converted by the mere presence of Galpha(s)QL to resemble that obtained with cicaprost in HEL cells . In addition , the restoration of the Galpha(16)-mediated inhibitor-sensitivity upon cicaprost induction in Galpha(s)-knocked down HEL cells again verified the pivotal role of G(s) signal . Taken together , our observations illustrate that co-stimulation of G(s) and G(q) can result in the fine-tuning of P40763 activation status , and this may provide the basis for cell type-specific responses following activation of hIP . The evolving genetic foundations of eating disorders . Data described earlier are clear in establishing a role for genes in the development of eating abnormalities . Estimates from the most rigorous studies suggest that more than 50 % of the variance in eating disorders and disordered eating behaviors can be accounted for by genetic effects . These high estimates indicate a need for studies identifying the specific genes contributing to this large proportion of variance . Twin and family studies suggest that several heritable characteristics that are commonly comorbid with AN and BN may share genetic transmission with these disorders , including anxiety disorders or traits , body weight , and possibly major depression . Moreover , some developmental research suggests that the genes involved in ovarian hormones or the genes that these steroids affect also may be genetically linked to eating abnormalities . Molecular genetic research of these disorders is in its infant stages . However , promising areas for future research have already been identified ( e.g. , 5- Q13049 receptor gene , P25874 -2/ P25874 -3 gene , and estrogen receptor beta gene ) , and several large-scale linkage and association studies are underway . These studies likely will provide invaluable information regarding the appropriate phenotypes to be included in genetic studies and the genes with the most influence on the development of these disorders . Menstrual cycle-dependent febrile episode mediated by sequence-specific repression of poly(ADP-ribose) polymerase-1 on the transcription of the human serotonin receptor 1A gene . The serotonin receptor 1A ( encoded by the P08908 gene ) plays a critical role in serotonergic transmission and was linked with many human diseases . A 33-year-old woman with rare menstrual cycle-dependent fever showed abnormal estrogen profile and responded well to the P08908 agonist buspirone , suggesting that her fevers were allied to estrogen-related P08908 deficiency . We identified an adenine deletion 480-bases upstream of the translation start site ( i.e. , -480delA ) of P08908 in this patient . To determine the underlying mechanism of -480delA-mediated P08908 deficiency , we first showed that P08908 -480 region can be bound by multiple nuclear protein(s) . We then identified poly(ADP-ribose) polymerase ( P09874 ) as one of the proteins that binds to P08908 -480 region . Using P09874 overexpression and knockdown , our data demonstrated that P09874 represses P08908 transcription . Furthermore , P08908 -480delA promoter possesses increased interaction with P09874 and caused an additional reduction in transcription . Finally , 17β-estradiol administration further reduced transcription associated with the mutant promoter . Altogether , these data suggest that estrogen-induced hyperactivity of P08908 mutant promoter causes the reduction of P08908 mRNA and leads to the disruption of P08908 -mediate hypothermic regulation . This is the first report of P08908 mutation underlying menstrual cycle-dependent febrile episodes , and implies that similar " febrile episode " cases may also result from the dysfunction of serotonin transmission . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . Clinical outcome and cyclo-oxygenase-2 expression in five dogs with solar dermatitis/actinic keratosis treated with firocoxib . BACKGROUND : The conversion of arachidonic acid into prostaglandin is catalysed by the cyclo-oxygenases ( P23219 / P35354 ) . Several studies indicate that P35354 is overexpressed in actinic keratosis in humans and dogs . DB09217 is a P35354 -selective inhibitor that blocks the biochemical activity of P35354 . HYPOTHESIS/OBJECTIVES : To evaluate the efficacy of firocoxib ( 5 mg/kg orally once daily ) for the treatment of dogs with solar dermatitis/actinic keratosis . METHODS : DB09217 5 mg/kg was given orally once daily for 180 days to five dogs with clinical signs and histopathological lesions consistent with solar dermatitis/actinic keratosis . On days 0 , 50 and 180 , the severity of erythema , skin shine , induration and the number of comedones were evaluated by a clinical scoring system . On the same days , samples were collected for histopathology from ' target lesions ' and P35354 expression was evaluated by immunohistochemistry . RESULTS : The clinical follow-up showed that four of five dogs improved with the treatment ; improvement in terms of histological findings was correlated with the regularization of the epidermal proliferation rather than the recovery of dermal changes . CONCLUSIONS AND CLINICAL IMPORTANCE : A role for P35354 might thus be hypothesized in the pathogenesis of canine solar dermatitis . DB06273 in pediatric rheumatology : the clinical experience . During the last two decades , clinical use of novel biological therapy has led to increased mechanistic understanding of complex rheumatological diseases . Conversely , basic and translational studies have led to development of new and varied therapeutic agents . These new medications which " target " specific steps in one or more immune pathways have the potential to control disease symptoms , improve quality of life and long-term prognosis , and perhaps in some , restore immunological tolerance . Use of these agents in clinical trials , combined with post-marketing surveillance , has revealed both the benefits and the undesirable side-effects of biological disease-modifying anti-rheumatic drugs ( DMARDs ) . In this review we focus on the use of tocilizumab , a monoclonal antibody directed against the P05231 receptor ( P08887 ) , which potently inhibits P05231 / P08887 signaling . 5- Q13049 receptor induces P29323 phosphorylation and proliferation through ADAM-17 tumor necrosis factor-alpha-converting enzyme ( P78536 ) activation and heparin-bound epidermal growth factor-like growth factor ( HB- P01133 ) shedding in mesangial cells . In this study , we present multiple lines of evidence to support a critical role for heparin-bound P01133 ( epidermal growth factor ) -like growth factor ( HB- P01133 ) and tumor necrosis factor-alpha-converting enzyme ( P78536 ) ( P78536 ) in the transactivation of P01133 receptor ( P00533 ) , P29323 phosphorylation , and cellular proliferation induced by the 5-HT(2A) receptor in renal mesangial cells . 5-hydroxy-tryptamine ( 5-HT ) resulted in rapid activation of P78536 , HB- P01133 shedding , P00533 activation , P29323 phosphorylation , and longer term increases in DNA content in mesangial cells . P29323 phosphorylation was attenuated by 1 ) neutralizing P00533 antibodies and the P00533 kinase inhibitor , AG1478 , 2 ) neutralizing HB- P01133 , but not amphiregulin , antibodies , heparin , or CM197 , and 3 ) pharmacological inhibitors of matrix-degrading metalloproteinases or P78536 small interfering RNA . Exogenously administered HB- P01133 stimulated P29323 phosphorylation . Additionally , P78536 was co-immunoprecipitated with HB- P01133 . Small interfering RNA against P78536 also blocked 5-HT-induced increases in P29323 phosphorylation , HB- P01133 shedding , and DNA content . In aggregate , this work supports a pathway map that can be depicted as follows : 5-HT --> 5-HT(2A) receptor --> P78536 --> HB- P01133 shedding --> P00533 --> P29323 --> increased DNA content . To our knowledge , this is the first time that P78536 has been implicated in 5-HT-induced P00533 transactivation or in proliferation induced by a G protein-coupled receptor in native cells in culture . Signaling pathways mediating induction of the early response genes prostaglandin G/H synthase-2 and egr-1 by serotonin via 5- Q13049 receptors . Signaling pathways responsible for serotonin ( 5-HT ) -mediated induction of early response genes prostaglandin G/H synthase-2 ( P35354 , cyclooxygenase-2 ) and egr-1 were investigated in rat mesangial cells . Gene induction by 5-HT was dependent on 5- Q13049 receptors that were pertussis toxin insensitive indicating coupling to a G-protein of the Gq family . Binding of 5-HT to this receptor activates phosphatidylinositol-specific phospholipase C ( P98160 ) and release of Ca2+ from internal stores , but this activation was not related to P35354 mRNA expression . Similarly , P19957 kinase was not involved in 5-HT signaling . Instead , inhibition of phosphatidylcholine-specific P98160 interfered with P35354 and egr-1 mRNA induction , suggesting this enzyme as a link between 5- Q13049 receptors and protein kinase C , an essential part of 5-HT-mediated signaling . The Q96HU1 kinase pathway was identified as common signaling pathway of 5-HT or phorbol ester-induced gene expression . Increase of intracellular DB02527 by forskolin or dibutyryl DB02527 did not induce P35354 or egr-1 mRNA expression by itself , but strongly inhibited 5-HT-mediated mRNA induction . P35354 mRNA and protein induction by 5-HT was also abolished by chelation of Ca2+ ions by EGTA , suggesting involvement of Ca2+-dependent enzymes . In contrast , egr-1 mRNA expression was superinduced in the presence of EGTA . Induction of Egr-1 protein was not changed by EGTA hinting to Ca2+-sensitive posttranscriptional steps . Activation of the Gq-coupled 5- Q13049 receptor thus leads to the expression of the early response genes P35354 and egr-1 , using common as well as differing signaling elements that allow differential regulation of the expression of these genes that are functionally related to renal hemodynamics and proliferation of mesangial cells , respectively .

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Participation of a cholinergic mechanism in 5-hydroxytryptamine (5-HT)3 and Q13639 receptor-mediated stimulation of gastric emptying in rats . The participation of a cholinergic mechanism in 5-hydroxytryptamine (5-HT)3 and Q13639 receptor-mediated stimulation of gastric emptying in rats was investigated . The selective 5- Q9H205 receptor antagonists ramosetron ( YM060 , 0.1-10 micrograms/kg i.v. ) and ondansetron ( 1-100 micrograms/kg i.v. ) dose-dependently enhanced the gastric emptying of glass beads in rats . The Q13639 receptor agonist 5-methoxytryptamine ( 5- P38646 , 1 mg/kg s.c. ) and substituted benzamides ( Q13639 receptor agonist/5- Q9H205 receptor antagonists ) cisapride ( 1-10 mg/kg s.c. ) and zacopride ( 1-1000 micrograms/kg s.c. ) produced significant gastroprokinetic responses in rats . The substituted benzamide-induced gastroprokinetic responses were inhibited by a high dose of tropisetron ( 10 mg/kg s.c. ) , a 5- Q9H205 and Q13639 receptor antagonist , and partially inhibited by GR113808 ( [1-[2-[(methylsulfonyl)amino]ethyl]-4-piperidyl]methyl 1-methyl-1H-indole-3-carboxylate , 1 mg/kg s.c. ) , a selective Q13639 receptor antagonist . On the other hand , the 5- P38646 -induced gastroprokinetic response was almost completely inhibited by GR113808 . In contrast , tetrodotoxin ( TTX , 0.1-10 micrograms/kg s.c. ) and atropine ( 1-1000 micrograms/kg s.c. ) dose-dependently inhibited gastric emptying . The enhancement of gastric emptying induced by selective 5- Q9H205 receptor antagonists , substituted benzamides and 5- P38646 was inhibited by TTX ( 10 micrograms/kg s.c. ) and by atropine ( 1 mg/kg s.c. ) . These results suggest that substituted benzamides stimulated gastric emptying partly due to their Q13639 receptor agonistic properties , and that both 5- Q9H205 receptor antagonism and Q13639 receptor agonism stimulated the gastric emptying in rats . It is also suggested that a cholinergic mechanism participates in the 5- Q9H205 and Q13639 receptor-mediated stimulation of gastric emptying in rats . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Newborn Analgesia Mediated by DB00107 during Delivery . The mechanisms controlling pain in newborns during delivery are poorly understood . We explored the hypothesis that oxytocin , an essential hormone for labor and a powerful neuromodulator , exerts analgesic actions on newborns during delivery . Using a thermal tail-flick assay , we report that pain sensitivity is two-fold lower in rat pups immediately after birth than 2 days later . P30559 antagonists strongly enhanced pain sensitivity in newborn , but not in 2-day-old rats , whereas oxytocin reduced pain at both ages suggesting an endogenous analgesia by oxytocin during delivery . Similar analgesic effects of oxytocin , measured as attenuation of pain-vocalization induced by electrical whisker pad stimulation , were also observed in decerebrated newborns . DB00107 reduced GABA-evoked calcium responses and depolarizing GABA driving force in isolated neonatal trigeminal neurons suggesting that oxytocin effects are mediated by alterations of intracellular chloride . Unlike GABA signaling , oxytocin did not affect responses mediated by P56373 and Q8NER1 receptors . In keeping with a GABAergic mechanism , reduction of intracellular chloride by the diuretic P55011 chloride co-transporter antagonist bumetanide mimicked the analgesic actions of oxytocin and its effects on GABA responses in nociceptive neurons . Therefore , endogenous oxytocin exerts an analgesic action in newborn pups that involves a reduction of the depolarizing action of GABA on nociceptive neurons . Therefore , the same hormone that triggers delivery also acts as a natural pain killer revealing a novel facet of the protective actions of oxytocin in the fetus at birth . Effects of thioacetamide-induced hepatic failure on the N-methyl-D-aspartate receptor complex in the rat cerebral cortex , striatum , and hippocampus . Binding of different ligands and expression of receptor subunit mRNAs . Hepatic encephalopathy ( HE ) is characterized by symptoms pointing at disturbances in glutamatergic neurotransmission in the brain , particularly in the striatum . The binding parameters of ligands specific for different recognition sites in the N-methyl-D-aspartate ( DB01221 ) receptor complex and the distribution of the receptor subunit mRNAs ( Q9UHB4 , Q12879 -D ) were assessed in rats with acute HE induced with a hepatotoxin , thioacetamide ( TAA ) . The binding of : 1 . L-[3H]glutamate ( DB01221 -displaceable ) ; 2 . [3H]dizocilpine and N-(1-[2-thienyl]-cyclohexyl) [3H]piperidine ( [3H] DB01520 ) ; and 3 . The coactivator site agonist [3H]glycine was assayed in purified membranes of the cerebral cortex , hippocampus , and striatum . In HE rats , Bmax of DB01221 -displaceable glutamate binding was increased in the cerebral cortex and hippocampus , but slightly decreased in the striatum . In this region , the binding affinity was also slightly increased . In HE , Bmax of [3H]dizocilpine binding was unchanged in the striatum and cerebral cortex , but substantially decreased in the hippocampus . Pretreatment with phorbol ester enhanced the binding of dizocilpine more in HE than in control rats . Bmax of [3H] DB01520 binding was decreased in the cerebral cortex and striatum , but increased in the hippocampus . The different responses of these two phencyclidine site antagonists to HE may be indicative of a conformational change within the ion channel and/or the presence of microdomains reacting differently to extrinsic factors . HE did not affect glycine binding , but potentiated the maximal stimulation of [3H]dizocilpine binding by glycine in the cerebral cortex . The results emphasize the brain region and domain specificity of the responses of the DB01221 receptor complex to HE . P30047 -dependent and -independent inhibitors of P30793 . P30047 ( P30047 ) mediates the feedback inhibition of P30793 activity by ( 6R ) -L-erythro- DB00360 ( BH4 ) through protein complex formation . Since guanine and BH4 have a common pyrimidine ring structure , we examined the inhibitory effect of guanine and its analogs on the enzyme activity . DB02377 , 8-hydroxyguanine , 8-methylguanine , and 8-bromoguanine inhibited the enzyme activity in a P30047 -dependent and pH-dependent manner and induced complex formation between P30793 and P30047 . The type of inhibition by this group is a mixed type . All these properties were shared with BH4 . In striking contrast , inhibition by DB01667 and 8-mercaptoguanine was P30047 -independent and pH-independent . The type of inhibition by DB01667 and 8-mercaptoguanine was a competitive type . The two compounds did not induce complex formation between the enzyme and P30047 . These results demonstrate that guanine compounds of the first group bind to the BH4-binding site of the P30793 / P30047 complex , whereas DB01667 and 8-mercaptoguanine bind to the active site of the enzyme . Finally , the possible implications in Lesch-Nyhan syndrome and Parkinson diseases of the inhibition of P30793 by guanine and 8-hydroxyguanine are discussed . Sequential treatment with flavopiridol synergistically enhances pyrrolo-1,5-benzoxazepine-induced apoptosis in human chronic myeloid leukaemia cells including those resistant to imatinib treatment . The Bcr-Abl kinase inhibitor , imatinib mesylate , is the front line treatment for chronic myeloid leukaemia ( CML ) , but the emergence of imatinib resistance has led to the search for alternative drug treatments and the examination of combination therapies to overcome imatinib resistance . The pro-apoptotic PBOX compounds are a recently developed novel series of microtubule targeting agents ( MTAs ) that depolymerise tubulin . Recent data demonstrating enhanced MTA-induced tumour cell apoptosis upon combination with the cyclin dependent kinase ( CDK ) -1 inhibitor flavopiridol prompted us to examine whether this compound could similarly enhance the effect of the PBOX compounds . We thus characterised the apoptotic and cell cycle events associated with combination therapy of the PBOX compounds and flavopiridol and results showed a sequence dependent , synergistic enhancement of apoptosis in CML cells including those expressing the imatinib-resistant T315I mutant . DB03496 reduced the number of polyploid cells formed in response to PBOX treatment but only to a small extent , suggesting that inhibition of endoreplication was unlikely to play a major role in the mechanism by which flavopiridol synergistically enhanced PBOX-induced apoptosis . The addition of flavopiridol following PBOX-6 treatment did however result in an accelerated exit from the G2/M transition accompanied by an enhanced downregulation and deactivation of the P06493 /cyclin B1 complex and an enhanced degradation of the inhibitor of apoptosis protein ( IAP ) survivin . In conclusion , results from this study highlight the potential of these novel series of PBOX compounds , alone or in sequential combination with flavopiridol , as an effective therapy against CML . The role of endothelium-derived hyperpolarizing factor and cyclooxygenase pathways in the inhibitory serotonergic response to the pressor effect elicited by sympathetic stimulation in chronic sarpogrelate treated rats . We have demonstrated that the antagonism of 5-HT2 receptors produces an enhancement of serotonergic sympathoinhibitory effect by P28221 and P34969 activation . The aim of this work was to determine mechanisms involved in the 5-hydroxytriptaminergic inhibitory action on the pressor responses elicited by sympathostimulation in pithed rats treated with a 5-HT2 receptor blocker . The blockade of 5-HT2 receptors was induced by orally sarpogrelate treatment ( 30 mg/kg/day ) . Two weeks later , animals were anaesthetized and pithed . A bolus injection of 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one ( ODQ ) ( 10 µg/kg ) , a guanylyl cyclase inhibitor , or indomethacin ( 2mg/kg ) , a non-selective P36551 inhibitor , prior to the infusion of ( 2S ) (+)-5-(1,3,5-trimethylpyrazol-4-yl)-2-(dimethylamino)tetralin , AS-19 ( 5 µg/kg/min ) were not able to abolish its inhibitory action . However , i.v. administration of glibenclamide ( 20mg/kg ) , a blocker of DB00171 -sensitive K(+) channels , completely reversed AS-19 sympathoinhibitory action . The inhibitory effect of 2-[5-[3-(4-methylsulfonylamino)benzyl-1,2,4-oxadiazol-5-yl]-1H-indol-3-yl]ethanamine , L-694,247 ( 5 µg/kg/min ) was abolished by indomethacin , whereas pretreatment with ODQ had no effect . DB04743 ( 3mg/kg ) , a P35354 inhibitor , completely reversed the inhibitory action of L-694,247 , whereas 1- [ [ 4,5-bis (4-methoxyphenyl)-2-thiazolyl ] carbonyl ] -4-methylpiperazine hydrochloride ( FR122047 ) ( 3mg/kg ) , a P23219 inhibitor , partially blocked this action . The sympathoinhibition by 5-HT ( 20 µg/kg/min ) could not be elicited after i.v. treatment with indomethacin plus glibenclamide . In conclusion , these results suggest that in chronic sarpogrelate-treated rats , the inhibitory serotonergic effect of the pressor responses induced by electrical stimulation of the sympathetic outflow via P34969 and P28221 receptor activation is mediated by KATP channel-mediated smooth muscle hyperpolarization and the P36551 pathway , respectively . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . [ Leukemia- and lymphoma-associated flow cytometric , cytogenetic , and molecular genetic aberrations in healthy individuals ] . Most leukemia and lymphoma cases are characterized by specific flow cytometric , cytogenetic and molecular genetic aberrations , which can also be detected in healthy individuals in some cases . The authors review the literature concerning monoclonal B-cell lymphocytosis , and the occurrence of chromosomal translocations t(14;18) and t(11;14) , P06748 - Q9UM73 fusion gene , O60674 V617F mutation , P11274 - P00519 fusion gene , P41212 - Q01196 ( P41212 - Q01196 ) , Q03164 - P51825 and P29590 - P10276 fusion gene in healthy individuals . At present , we do not know the importance of these aberrations . From the authors review it is evident that this phenomenon has both theoretical and practical ( diagnostic , prognostic , and therapeutic ) significance . Characterization of a novel Q13639 receptor antagonist of the azabicycloalkyl benzimidazolone class : DAU 6285 . Three chemical classes of serotonin Q13639 receptor agonists have been identified so far : 5-substituted indoles ( e.g. 5-HT ) , benzamides ( e.g. renzapride ) and benzimidazolones ( e.g. BIMU 8 ) . In a search for Q13639 receptor antagonists , we have discovered that the benzimidazolone derivative DAU 6285 ( for structure see text ) , is 3-5 times more potent than tropisetron in blocking 5-HT , renzapride and BIMU 8 induced stimulation of adenylate cyclase activity in mouse embryo colliculi neurons . Schild plot analysis yielded Ki values of 220 , 181 and 255 nmol/l , respectively . In addition , DAU 6285 showed poor activity as a 5- Q9H205 receptor ligand with respect to tropisetron , as demonstrated by in vitro binding studies ( Ki , 322 vs 2.8 nmol/l ) and by its antagonistic activity in the Bezold-Jarisch reflex test ( ID50 , 231 vs 0.5 micrograms/kg , i.v. ) . No significant binding ( Ki greater than 10 mumol/l ) of DAU 6285 to serotonergic P08908 , P28222 , P28335 , P28221 , and 5-HT2 receptors as well as to adrenergic alpha 1 , alpha 2 , dopaminergic D1 , D2 or muscarinic M1-M3 receptor subtypes was found . The data indicate that DAU 6285 has a somewhat higher affinity than tropisetron for Q13639 receptors , a property confirmed in functional tests , and much lower affinity than tropisetron for 5- Q9H205 receptors . The compound represents a new interesting tool for investigating the pharmacological and physiological properties of Q13639 receptors . [ Effects of various growth factors on the growth of trophoblast cells in long-term culture ] . Trophoblasts taken from placental tissue of the 1st trimester and molar tissue , and P30793 -1 ( gestational choriocarcinoma cell line ) cells were cultured in collagen coated dishes . The medium used was a mixture of DME and Ham 's F-12 ( 1:1 ) , containing P01133 , PDGF , insulin , GM- P04141 , IL-1 , -2 , -3 , PGE1 and DB00917 in various concentrations . 3H-TdR uptake of the cultured cells was measured as a marker of cell growth . The growth of normal trophoblasts was enhanced by PDGF or insulin , and remarkably by P01133 + PDGF + insulin + GM- P04141 . The growth of molar trophoblasts was accelerated by P01133 or insulin . Under these culture conditions , normal trophoblasts have been successfully cultured and maintained for 12 months and molar trophoblasts for 9 months to date . The cultured cells were identified with trophoblasts by immunohistochemical staining with P62158 5.2 monoclonal antibody . No effect of growth factors was observed in P30793 -1 cells . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . In vitro modulation of the interaction between Q9ULX6 and LAP2beta by DB02527 signaling . The nuclear envelope mediates key functions by interacting with chromatin . We recently reported an interaction between the chromatin- and nuclear matrix-associated protein Q9ULX6 and the inner nuclear membrane integral protein LAP2beta , implicated in initiation of DNA replication ( Martins et al. ( 2003 ) J. Cell Biol. 160 , 177-188 ) . Here , we show that in vitro , interaction between Q9ULX6 and LAP2beta is modulated by DB02527 signaling via PKA . Exposure of an anti- Q9ULX6 immune precipitate from interphase HeLa cells to a mitotic extract promotes DB00171 -dependent release of LAP2beta from the Q9ULX6 complex . This coincides with DB00133 and DB00156 phosphorylation of Q9ULX6 and LAP2beta . Inhibition of PKA with PKI abolishes phosphorylation of Q9ULX6 and dissociation of LAP2beta from Q9ULX6 , although LAPbeta remains phosphorylated . Antagonizing DB02527 signaling in mitotic extract also abolishes the release of LAP2beta from Q9ULX6 ; however , disrupting PKA anchoring to A-kinase anchoring proteins has no effect . Inhibition of CDK activity in the extract greatly reduces LAP2beta phosphorylation but does not prevent LAP2beta release from Q9ULX6 . Inhibition of PKC , Q96HU1 kinase , or P62158 kinase II does not affect mitotic extract-induced dissociation of LAP2beta from Q9ULX6 . PKA phosphorylates Q9ULX6 but not LAP2beta in vitro and elicits a release of LAP2beta from Q9ULX6 . P06493 or PKC phosphorylates LAP2beta within the Q9ULX6 complex , but neither kinase induces LAP2beta release . Our results indicate that in vitro , the interaction between Q9ULX6 and LAP2beta is influenced by a PKA-mediated phosphorylation of Q9ULX6 rather than by P06493 - or PKC-mediated phosphorylation of LAP2beta . This suggests an additional level of regulation of a chromatin-nuclear envelope interaction in dividing cells . DB01233 does not increase gastric muscle contractility in newborn rats . Feeding intolerance resulting from delayed gastric emptying is common in premature neonates . DB01233 ( MCP ) , the most frequently used prokinetic drug in neonates , enhances gastric muscle contractility through inhibition of dopamine receptors . Although its therapeutic benefit is established in adults , limited data are available to support its clinical use in infants . Hypothesizing that developmentally dependent differences are present , we comparatively evaluated the effect of MCP on fundus muscle contractility in newborn , juvenile , and adult rats . The muscle strips were either contracted with electrical field stimulation ( O43281 ) to induce cholinergic nerve-mediated acetylcholine release or carbachol , a cholinergic agonist acting directly on the muscarinic receptor . Although in adult rats MCP increased O43281 -induced contraction by 294 ± 122 % of control ( P < 0.01 ) , no significant effect was observed in newborn fundic muscle . MCP had no effect on the magnitude of the carbachol-induced and/or bethanechol-induced gastric muscle contraction at any age . In response to dopamine , an 80.7 ± 5.3 % relaxation of adult fundic muscle was observed , compared with only a 8.4 ± 8.7 % response in newborn tissue ( P < 0.01 ) . P14416 expression was scant in neonates and significantly increased in adult gastric tissue ( P < 0.01 ) . In conclusion , the lack of MCP effect on the newborn fundic muscle contraction potential relates to developmental differences in dopamine D2 receptor expression . To the extent that these novel data can be extrapolated to neonates , the therapeutic value of MCP as a prokinetic agent early in life requires further evaluation . High-performance liquid chromatographic radioenzymatic assay for plasma catecholamines . A new assay method for plasma CA 's requiring only 50 microliter has been developed , which uses HPLC . The NE , D , and E compounds found in plasma are radioactively o-methylated with 3H- DB00118 by the reaction of P21964 . The reaction is terminated and a standard mixture of nonradioactive o-methylated analogues of NE , D , and E is addded to act as a carrier . Following separation by HPLC , the NMN , 3- P38646 , and MN radioactive peaks are collected which represent NE , D , and E , respectively . Then MNM and MN are oxidized to vanillin , and 3- P38646 is acetylated . The products are subsequently separated by solvent extraction . This is necessary in order to avoid high radioactive blanks and to allow quantitation of the radioactivity by liquid scintillation spectrometry . The mean supine levels of NE , D , and E in normal subjects were respectively 182 , 33 , and 87 pg/ml of plasma . Similar assays on patients with pheochromocytoma revealed 797 , 80 , and 470 pg/ml . Vascular endothelial growth factor signaling is required for the behavioral actions of antidepressant treatment : pharmacological and cellular characterization . This study extends earlier work on the role of vascular endothelial growth factor ( P15692 ) in the actions of antidepressant treatment in two key areas . First , by determining the requirement for P15692 in the actions of a 5-HT selective reuptake inhibitor ( SSRI ) , fluoxetine in behavioral models of depression/antidepressant response ; and second , by examining the role of the P08908 receptor subtype in the regulation of P15692 , and the cellular localization of antidepressant regulation of P15692 expression . The results show that pharmacological inhibition of P15692 receptor signaling blocks the behavioral actions of fluoxetine in rats subjected to chronic unpredictable stress . Infusions of SU5416 or SU1498 , two structurally dissimilar inhibitors of P15692 -Flk-1 receptor signaling , block the antidepressant effects of fluoxetine on sucrose preference , immobility in the forced swim test , and latency to feed in the novelty suppressed feeding paradigm . We also show that activation of P08908 receptors is sufficient to induce P15692 expression and that a P08908 antagonist blocks both the increase in P15692 and behavioral effects induced by fluoxetine . Finally , double labeling studies show that chronic fluoxetine administration increases P15692 expression in both neurons and endothelial cells in the hippocampus . Taken together these studies show that P15692 is necessary for the behavioral effects of the SSRI fluoxetine , as well as norepinephrine selective reuptake inhibitor , and that these effects may be mediated by P08908 receptors located on neurons and endothelial cells . DB06802 , a unique nonsteroidal prodrug with potential utility in the treatment of trauma-induced ocular inflammation : I . Assessment of anti-inflammatory efficacy . DB06802 , the amide analog of 2-amino-3-benzoylbenzeneacetic acid ( amfenac ) , was examined in preclinical models for its potential utility as a topical ocular anti-inflammatory agent . Diclofenac was selected as the reference compound . In contrast to diclofenac ( IC50 = 0.12 microM ) , nepafenac exhibited only weak P23219 inhibitory activity ( IC50 = 64.3 microM ) . However , amfenac was a potent inhibitor of both P23219 ( IC50 = 0.25 microM ) and P35354 activity ( IC50 = 0.15 microM ) . Ex vivo , a single topical ocular dose of nepafenac ( 0.1 % ) inhibited prostaglandin synthesis in the iris/ciliary body ( 85-95 % ) and the retina/choroid ( 55 % ) . These levels of inhibition were sustained for 6 h in the iris/ciliary body and 4 h in the retina/choroid . Diclofenac ( 0.1 % ) suppressed iris/ciliary body prostaglandin synthesis ( 100 % ) for only 20 min , with 75 % recovery observed within 6 h following topical dosing . Diclofenac 's inhibition of prostaglandin synthesis in the retina/choroid was minimal . DB06802 's inhibitory efficacy and longer duration of action was confirmed in a trauma-induced rabbit model of acute ocular inflammation monitoring protein or DB00917 accumulation in aqueous humor . Results warrant further assessment of nepafenac 's topical ocular efficacy in the treatment of postoperative ocular pain , inflammation , and posterior segment edema . DB00107 microinjected into the central amygdaloid nuclei exerts anti-aggressive effects in male rats . We recently demonstrated that acute and chronic intracerebroventricular enhancement of brain P01178 levels induces potent anti-aggressive and pro-social explorative effects during social challenges . However , the exact anatomical location in the brain where P01178 exerts its action is still elusive . In the present study , we targeted two critical brain areas , i.e. the central amygdala ( CeA ) and the dorsal raphe ( DR ) , both containing high levels of P01178 receptors ( OXTRs ) and constituting important nodes of the neural circuitry related to aggression . Behavioral effects of local micro-infusion of P01178 and P30559 antagonist , L368.899 , ( alone and combined ) were evaluated in resident male rats during confrontations with an unfamiliar male intruder . Our results show that P01178 microinjected into the CeA markedly reduced resident 's offensive behavior and facilitated social exploration , without affecting other non-aggressive behaviors . The receptor specificity of the behavioral effects was verified when a micro-infusion of a selective P30559 antagonist nullified the changes . Pharmacological blockade of CeA OXTRs per se was without clear behavioral effects suggesting that endogenous P01178 within the CeA does not play a major inhibitory role on offensiveness . Anatomical specificity was also supported by the absence of relevant behavioral effects when P01178 was microinjected into more medial sub-regions of the amygdala . Likewise , within the DR neither P01178 nor P30559 exerted significant effects on offensive aggression , while microinjection of the P08908 autoreceptor agonist in this region significantly suppressed aggression . In conclusion , our results point at the CeA as an important brain site of action for the anti-aggressive and pro-social explorative effects induced by exogenous enhancement of brain P01178 levels . Retinoic acid and synthetic analogs differentially activate retinoic acid receptor dependent transcription . We have developed an assay where the potency of retinoids in retinoic acid receptor ( RAR ) mediated transcriptional activation can be rapidly evaluated . In this assay hRAR-alpha , hRAR-beta and hRAR-gamma were expressed in CV-1 cells together with a reporter gene containing a retinoic acid responsive element ( TRE3-tk-CAT ) . Concentrations required to obtain half-maximum induction ( ED50 ) of CAT-activity were determined for several retinoids , e.g. , all-trans-retinoic acid ( RA ) , 13-cis-retinoic acid ( 13- DB00982 ) , arotinoid acid ( DB02877 ) and m-carboxy-arotinoid acid ( m-carboxy- DB02877 , an inactive arotinoid analog ) . The ED50 values for RA decreased in the order of P10276 ( 24 nM ) greater than P10826 ( 4.0 nM ) greater than P13631 ( 1.3 nM ) , while the ED50 values for DB02877 and 13- DB00982 decreased in the order of P10276 ( 6.5 nM , 190 nM ) greater than P13631 ( 2.3 nM , 140 nM ) greater than P10826 ( 0.6 nM , 43 nM ) , respectively . No significant inductions were obtained when cells were treated with m-carboxy- DB02877 , even at 10 microM concentrations . The fold induction of CAT-activity for all compounds tested decreased in the order of P10276 greater than P10826 greater than P13631 . Emergence of motor circuit activity . In the developing nervous system , ordered neuronal activity patterns can occur even in the absence of sensory input and to investigate how these arise , we have used the model system of the embryonic chicken spinal motor circuit , focusing on motor neurons of the lateral motor column ( O15467 ) . At the earliest stages of their molecular differentiation , we can detect differences between medial and lateral O15467 neurons in terms of expression of neurotransmitter receptor subunits , including P30532 , P36544 , Q12879 , P39086 , P08908 and P28222 , as well as the Q9H2X9 transporter . Using patch-clamp recordings we also demonstrate that medial and lateral O15467 motor neurons have subtly different activity patterns that reflect the differential expression of neurotransmitter receptor subunits . Using a combination of patch-clamp recordings in single neurons and calcium-imaging of motor neuron populations , we demonstrate that inhibition of nicotinic , muscarinic or GABA-ergic activity , has profound effects of motor circuit activity during the initial stages of neuromuscular junction formation . Finally , by analysing the activity of large populations of motor neurons at different developmental stages , we show that the asynchronous , disordered neuronal activity that occurs at early stages of circuit formation develops into organised , synchronous activity evident at the stage of O15467 neuron muscle innervation . In light of the considerable diversity of neurotransmitter receptor expression , activity patterns in the O15467 are surprisingly similar between neuronal types , however the emergence of patterned activity , in conjunction with the differential expression of transmitter systems likely leads to the development of near-mature patterns of locomotor activity by perinatal ages . Vascular endothelial growth factor trap-eye and trap technology : DB08885 from bench to bedside . Anti-vascular endothelial growth factor ( P15692 ) currently used to treat eye diseases have included monoclonal antibodies , antibody fragments , and an aptamer . A different method of achieving P15692 blockade in retinal diseases includes the concept of a cytokine trap . Cytokine traps technology are being evaluated for the treatment of various diseases that are driven by excessive cytokine levels . Traps consist of two extracellular cytokine receptor domains fused together to form a human immunoglobulin G ( IgG ) . DB08885 / P15692 trap-eye ( VTE ) is a soluble fusion protein , which combines ligand-binding elements taken from the extracellular components of P15692 receptors 1 and 2 fused to the Fc portion of IgG . This protein contains all human amino acid sequences , which minimizes the potential for immunogenicity in human patients . This review presents the latest data on VTE in regard to the pharmacokinetics , dosage and safety , preclinical and clinical experiences . Method of the literature search : A systematic search of the literature was conducted on PubMed , Scopus , and Google Scholar with no limitation on language or year of publication databases . It was oriented to articles published for VTE in preclinical and clinical studies and was focused on the pharmacokinetics , dosage and safety of VTE .

MH_train_1353

MH_train_1353

MH_train_1353

interacts_with DB00477?

multiple_choice

Q07869 gamma ligands , rosiglitazone and pioglitazone , inhibit P09038 - and P15692 -mediated angiogenesis . OBJECTIVE : To study the effect of peroxisome proliferator-activated receptor-gamma ( Q07869 gamma ) agonists , pioglitazone and rosiglitazone , on vascular endothelial growth factor ( P15692 ) - and basic fibroblast growth factor ( P09038 ) -induced angiogenesis and on endothelial cell migration . METHODS : Chick chorioallantoic membrane ( P62158 ) model was used to evaluate the efficacy of pioglitazone and rosiglitazone on P15692 - and P09038 -induced angiogenesis . In addition , the effect of pioglitazone and rosiglitazone on endothelial cell migration was evaluated using 8 mm pore filter to a feeder layer containing vitronectin as chemoattractant . RESULTS : Pioglitazone and rosiglitazone inhibited the pro-angiogenic effects of P09038 and P15692 in the P62158 model significantly ( P < 0.001 ) to the same extent . Endothelial cell migration was also inhibited by both pioglitazone and rosiglitazone ( P < 0.001 ) . CONCLUSIONS : These results suggest that Q07869 gamma ligands , pioglitazone and rosiglitazone , in addition to their important regulatory role in adipogenesis and inflammation , possess anti-angiogenic properties . Thus , Q07869 gamma ligands may be useful in the treatment of diabetic retinopathy , macular degeneration , and other ocular disorders and may lower the risk to develop cancer in diabetic patients . Intramural müllerian papilloma of the vagina . A 24-year-old pregnant woman was found to have a mass in the posterior vagina ; a partly cystic and partly solid lesion was excised . Histology showed a papillary mural neoplasm covered by intact squamous epithelium . Immunohistochemical staining showed diffuse positivity with P62158 5.2 , EP4 , P06731 , and P15941 and focal immunoreactivity with Q8WXI7 . This lesion was interpreted as an intramural mullerian papilloma , only the second such tumor described in the vagina and the first in an adult . DB00227 -stimulated superinduction of P16581 , P05362 and P19320 in P01375 activated human vascular endothelial cells . Inhibitors of P04035 ( statins ) reveal important pharmacological effects in addition to reducing the plasma LDL cholesterol level . In the pathogenesis of arteriosclerosis , transendothelial migration of various leukocytes including monocytes is a crucial step . We , therefore , investigated the expression of P16581 , intercellular cell adhesion molecule-1 ( P05362 ) and vascular cell adhesion molecule-1 ( P19320 ) in vascular endothelial cells as influenced by lovastatin . Human umbilical vein endothelial cells ( HUVECs ) express significant amounts of selectins and cell adhesion molecules ( CAMs ) within a few hours after stimulation with P01375 . This effect is potentiated by 100-200 % when the cells are pretreated with 0.1-2.5 microM lovastatin . The lovastatin-mediated increase in the cytoplasm and at the cell surface is dose-dependent and significant at lovastatin concentrations comparable to plasma levels in patients under lovastatin treatment . The lovastatin-potentiated increase of P16581 and CAMs is correlated with a corresponding increase of selectin- and P62158 -specific mRNA . We conclude that , in vivo , statin treatment could trigger an enhanced recruitment of macrophages that might support the cholesteryl ester efflux from the arteriosclerotic plaque . Proinflammatory cytokines promote glial heme oxygenase-1 expression and mitochondrial iron deposition : implications for multiple sclerosis . Proinflammatory cytokines , pathological iron deposition , and oxidative stress have been implicated in the pathogenesis of multiple sclerosis ( MS ) and experimental autoimmune encephalomyelitis ( EAE ) . P09601 mRNA levels and mitochondrial uptake of [(55)Fe]Cl(3)-derived iron were measured in rat astroglial cultures exposed to interleukin-1beta ( IL-1beta ) or tumor necrosis factor-alpha ( P01375 ) alone or in combination with the heme oxygenase-1 ( P09601 ) inhibitors , tin mesoporphyrin ( DB04912 ) or dexamthasone ( DEX ) , or interferon beta1b ( P27352 -beta ) . P09601 expression in astrocytes was evaluated by immunohistochemical staining of spinal cord tissue derived from MS and control subjects . IL-1beta or P01375 promoted sequestration of non-transferrin-derived (55)Fe by astroglial mitochondria . P09601 inhibitors , mitochondrial permeability transition pore ( P55157 ) blockers and antioxidants significantly attenuated cytokine-related mitochondrial iron sequestration in these cells . IFN-beta decreased P09601 expression and mitochondrial iron sequestration in IL-1beta- and P01375 -challenged astroglia . The percentage of astrocytes coexpressing P09601 in affected spinal cord from MS patients ( 57.3 % +/- 12.8 % ) was significantly greater ( p < 0.05 ) than in normal spinal cord derived from controls subjects ( 15.4 % +/- 8.4 % ) . P09601 is over-expressed in MS spinal cord astroglia and may promote mitochondrial iron deposition in MS plaques . In MS , IFN-beta may attenuate glial P09601 gene induction and aberrant mitochondrial iron deposition accruing from exposure to proinflammatory cytokines . Stimulation of type 1 and type 8 Ca2+/calmodulin-sensitive adenylyl cyclases by the Gs-coupled 5-hydroxytryptamine subtype 5-HT7A receptor . The neurotransmitter serotonin ( 5-hydroxytryptamine , 5-HT ) plays an important regulatory role in developing and adult nervous systems . With the exception of the 5- Q9H205 receptor , all of the cloned serotonin receptors belong to the G protein-coupled receptor superfamily . Subtypes P50406 and P34969 couple to stimulation of adenylyl cyclases through Gs and display high affinities for antipsychotic and antidepressant drugs . In the brain , mRNA for P50406 is found at high levels in the hippocampus , striatum , and nucleus accumbens . P34969 mRNA is most abundant in the hippocampus , neocortex , and hypothalamus . To better understand how serotonin might control DB02527 levels in the brain , we coexpressed P50406 or 5-HT7A receptors with specific isoforms of adenylyl cyclase in P29320 293 cells . The P50406 receptor functioned as a typical Gs-coupled receptor in that it stimulated O95622 , a Gs-sensitive adenylyl cyclase , but not Q99440 or P40145 , calmodulin ( P62158 ) -stimulated adenylyl cyclases that are not activated by Gs-coupled receptors in vivo . Surprisingly , serotonin activation of 5-HT7A stimulated Q99440 and P40145 by increasing intracellular Ca2+ . 5-HT also increased intracellular Ca2+ in primary neuron cultures . These data define a novel mechanism for the regulation of intracellular DB02527 by serotonin . Implantation of P15692 transfected preadipocytes improves vascularization of fibrin implants on the cylinder chorioallantoic membrane ( P62158 ) model . The successful substitution or augmentation of soft tissues by implantation of three dimensional cell constructs , consisting of human preadipocytes and fibrin glue as a carrier matrix , requires a rapid and homogeneous vascularization of the whole implant in order to provide a sufficient blood supply of centrally situated cells . Previous investigations have shown that under in vivo conditions primary human preadipocytes induce vascularization of fibrin matrices by secretion of several growth factors , such as P15692 and P09038 . The current study investigates whether vascularization of implants can be improved by transplantation of preadipocytes following transfection with a P15692 -vector . Transfection was performed by electroporation with an pCMX-GFP and pCMX-VEGF165 vector . Transfection efficiency ( GFP expression ) and P15692 expression were determined in vitro by FACS analysis and P15692 immunoassay , respectively . In vivo investigations to determine the vascularization of the implants were performed on the cylinder chorioallantoic membrane ( P62158 ) . Four million P15692 transfected cells were transferred within a fibrin matrix onto the P62158 on the 7(th) day of incubation and after 8 days the vascularization of the implant was histologically examined and evaluated by means of a computer-assisted image analysis program . Transfection of preadipocytes with the GFP vector by electroporation yielded transfection efficiencies between 12 % and 41 % of surviving cells . Results of the P15692 immunoassay demonstrated that P15692 expression was significantly higher following transfection . Investigations on the P62158 outlined a significantly higher rate of vascularization in the transfected vs. control population . Our investigations demonstrate that primary human preadipocytes can be successfully transfected by electroporation with a P15692 vector . The enhanced P15692 expression on transfected cells results in an increase of vascularization of the cell constructs on the P62158 . [ Functional characteristics of calcium-sensitive adenylyl cyclase of ciliate Tetrahymena pyriformis ] . DB01373 -sensitive forms of adenylyl cyclase ( AC ) were revealed in most vertebrates and invertebrates and also in some unicellular organisms , in particular ciliates . We have shown for the first time that calcium cations influence the AC activity of ciliate Tetrahymena pyriformis . These cations at the concentrations of 0.2-20 microM stimulated the enzyme activity , and maximum of catalytic effect was observed at 2 microM Ca2+ . DB01373 cations at a concentrations of 100 microM or higher inhibited the AC activity . P62158 antagonists W-5 and W-7 at the concentrations of 20-100 microM inhibited the catalytic effect induced by 5 microM Ca2+ and blocked the effect at higher concentrations of Ca2+ . DB00477 , another calmodulin antagonist , reduced Ca2+-stimulated AC activity only at the concentrations of 200-1000 microM . AC stimulating effects of serotonin , P01133 and DB02527 increased in the presence of 5 microM Ca2+ . AC stimulating effects of P01133 , DB02527 and insulin decreased in the presence of 100 microM Ca2+ , and AC stimulating effect of DB02527 decreased also in the presence of calmodulin antagonists ( 1 mM ) . At the same time , stimulating effect of D-glucose in the presence of Ca2+ and calmodulin antagonists did not change essentially . The data obtained speak in favor of the presence of calcium-sensitive forms of AC in ciliate T. pyriformis which mediate enzyme stimulation by P01133 , DB02527 , insulin , and serotonin . Biologic and immunologic therapies for ovarian cancer . Biologic therapy of ovarian cancer has been conducted using nonspecific biologic response modifiers , cytokines , monoclonal antibodies ( MAbs ) , vaccines , and gene therapy . Antibodies directed toward her2/neu have also been studied . Phase I and II gene therapy trials using adenoviral vectors containing a wild-type or modified p53 have shown that the treatment is well tolerated . Phase II and III trials are ongoing with MAbs directed against Q8WXI7 ( MAb DB04964 ) and an antibody directed against HMFG1 ( anti-HMFG1-yttrium-90-labeled antibody ) . The trials have shown that these agents are well tolerated and that immunologic responses occur , although the ultimate clinical value of these agents remains to be determined . Prolonged survival after MAb DB04964 treatment has been correlated with changes in several immune parameters , including human antimurine antibody , Ab2 , anti- Q8WXI7 antibody development , and induced T-cell immunity . Clinical trials using a MAb directed toward the encoded products of her2/neu have shown minimal activity against ovarian cancer in a phase I and II trial conducted by the Gynecologic Oncology Group . Cytokine therapies have been administered systemically and intraperitoneally . Intracavitary interferon alfa , interferon gamma , and interleukin-2 alone or in combination with cytotoxic therapy in phase I and II trials demonstrated intraperitoneal lymphoid cell stimulation and produced antitumor responses . A randomized trial of chemotherapy with or without interferon gamma in primary treatment produced a response and a progression-free survival advantage in the arm that incorporated the interferon gamma , without a statistically significant benefit in overall survival . A phase III study of interferon gamma in combination with first-line chemotherapy is currently ongoing . Nitrergic response to cyclophosphamide treatment in blood and bone marrow . Daily intraperitoneal injection of cyclophosphamide ( P15085 ) ( 50 mgkg(-1) of body weight ) for 5 days resulted in reduced levels of marrow and blood cellularity , which was most pronounced in 18 days post-treatment ( pt ) . On day 18 after P15085 treatment the enhancedlevels of nitric oxide ( NO ) precursors and metabolites ( L-arginine , L-citrulline , reactive nitrogen species ( RNS ) ) of marrow and blood cells ( platelet , neutrophil , lymphocyte and monocyte ) resulted from up-regulation of Ca(II)/calmodulin( P62158 )-independent " inducible " NO synthase ( P35228 ) , with a lessercontribution of Ca(II)/ P62158 -dependent " constitutive " P29474 isoforms to systemic NO.Biphasic response to P15085 of marrow nitrergic system , i.e. both P35228 and P29474 showed significantly depressed activities , as well as diminished levels of NO metabolites on day 9 pt , suggested that signals in addition to NO might be involved in P15085 -induced inhibition of hematopoesis , while a gradual increase of neutrophil and platelet NOS activity appeared to be contributed to a P15085 -induced development of granulopenia , thrombocytopenia and hemorrhage . The glial cell modulator and phosphodiesterase inhibitor , DB05066 ( ibudilast ) , attenuates prime- and stress-induced methamphetamine relapse . Stress and renewed contact with drug ( a " slip " ) have been linked to persisting relapse of methamphetamine abuse . Human brain microglial activation has been linked with methamphetamine abuse , and inhibitors of glial cell activation , certain phosphodiesterase ( PDE ) inhibitors , and glial cell derived neurotrophic factor ( P39905 ) have been reported to modulate drug abuse effects . Our objective was to determine whether the glial cell attenuator , 3-isobutyryl-2-isopropylpyrazolo-[1,5-a]pyridine ( DB05066 , ibudilast ) , a non-selective PDE inhibitor and promoter of P39905 , could reduce stress- and methamphetamine prime-induced reinstatement of methamphetamine-seeking behavior . Male Long-Evans hooded rats were trained to lever press reinforced with 0.1 mg/kg i.v. methamphetamine infusion according to fixed-ratio 1 ( FR1 ) reinforcement schedules during daily , 2-hour experimental sessions . After performance had stabilized , lever pressing was extinguished for 12 consecutive sessions and doses of 0 ( vehicle ) , 2.5 and 7.5 mg/kg DB05066 were then administered intraperitoneally b.i.d. on the last 2 days of extinction and then once on the testday to separate groups of 12 rats . During testing , the rats were given 15 min of intermittent footshock or a 1 mg/kg i.p. methamphetamine prime followed by a 2-hour reinstatement test session . DB05066 significantly reduced response levels of footshock-induced ( 2.5 and 7.5 mg/kg ) and prime-induced ( 7.5 mg/kg ) reinstatement of extinguished methamphetamine-maintained responding . DB05066 has properties consistent with the ability to attenuate relapse precipitated by stress and methamphetamine " slips " during abstinence . These results thus reinforce interest in atypical neurobiological mechanisms which could be exploited for developing novel medications for treating drug abuse disorders . DB06366 monotherapy after trastuzumab-based treatment and subsequent reintroduction of trastuzumab : activity and tolerability in patients with advanced human epidermal growth factor receptor 2-positive breast cancer . PURPOSE : The combination of pertuzumab and trastuzumab resulted in a clinical benefit rate ( P16152 ) of 50 % in patients with human epidermal growth factor receptor 2 ( P04626 ) -positive breast cancer whose disease progressed during prior trastuzumab-based therapy . To define whether this previously observed encouraging activity was a result of the combination of pertuzumab and trastuzumab or of pertuzumab alone , we recruited a third cohort of patients who received pertuzumab without trastuzumab . We then investigated the impact of reintroducing trastuzumab to patients whose disease progressed on pertuzumab monotherapy . PATIENTS AND METHODS : Twenty-nine patients with P04626 -positive breast cancer whose disease progressed during prior trastuzumab-based therapy received pertuzumab ( 840 mg loading dose , then 420 mg every 3 weeks ) until progressive disease or unacceptable toxicity . Seventeen patients with disease progression continued to receive pertuzumab ( at the same dose ) , with the addition of trastuzumab ( 4 mg/kg loading dose and then 2 mg/kg weekly or 8 mg/kg loading dose and then 6 mg/kg every 3 weeks ) . RESULTS : All 29 patients enrolled for pertuzumab monotherapy experienced disease progression . The objective response rate ( ORR ) and P16152 were 3.4 % and 10.3 % , respectively , during pertuzumab monotherapy . With the addition of trastuzumab , the ORR and P16152 were 17.6 % and 41.2 % , respectively . Progression-free survival was longer with combination therapy than pertuzumab monotherapy ( 17.4 v 7.1 weeks , respectively ) . Treatment was well tolerated with minimal cardiac dysfunction . CONCLUSION : Although pertuzumab has some activity in patients with P04626 -positive breast cancer that progressed during therapy with trastuzumab , the combination of pertuzumab and trastuzumab seems to be more active than monotherapy . DB06288 promotes cognitive flexibility in rats : the role of P34969 receptors . The antagonism of P34969 receptors may contribute to the antidepressant and procognitive actions of the atypical antipsychotic drug , amisulpride . It has been previously demonstrated that the selective P34969 receptor antagonist reversed restraint stress-induced cognitive impairments in a rat model of frontal-dependent attentional set-shifting task ( ASST ) . Therefore , the first aim of the present study was to assess the effectiveness of amisulpride against stress-evoked cognitive inflexibility . The second goal was to elucidate whether the pro-cognitive effect of amisulpride could be due to the compound 's action at P34969 receptors . Rats repeatedly exposed ( 1 h daily for 7 days ) to restraint stress demonstrated impaired performance on the extra-dimensional ( ED ) set-shifting stage of the ASST . DB06288 ( 3 mg/kg ) given to stressed rats 30 min before testing reversed this restraint-induced cognitive inflexibility and improved ED performance of the unstressed control group . The P34969 receptor agonist , AS19 ( 10 mg/kg ) , abolished the pro-cognitive efficacy of amisulpride ( 3 mg/kg ) . The present study suggests that the antagonism of P34969 receptors may contribute to the mechanisms underlining the pro-cognitive action of amisulpride . These results may have therapeutic implications in frontal-like deficits associated with stress-related disorders . Cloning of a calmodulin kinase I homologue from Schizosaccharomyces pombe . By using (35)S-labeled calmodulin ( P62158 ) , we have isolated a full-length cDNA clone expressing the Schizosaccharomyces pombe homologue of calmodulin kinase I ( CaMK-I ) , a gene we have named cmk1 . It has been previously been shown in mammals that CaMK-I is a member of a P62158 -dependent protein kinase cascade that ultimately regulates transcription factors such as P39905 and DB02527 -response element-binding protein . The cmk1 cDNA encodes a 335-amino acid protein with significant homology to mammalian CaMK-I , including a conserved sequence for phosphorylation by P62158 kinase kinase . We have expressed the cmk1 cDNA in bacteria and yeast , and we have shown that it is a P62158 -dependent protein kinase . A truncation mutant of cmk1 ( d320 ) failed to bind P62158 , indicating that the P62158 -binding domain is at the extreme C terminus of the protein . The mRNA for cmk1 is expressed in a cell cycle-dependent manner , peaking at or near the G(1)/S boundary . Overexpression of wild-type cmk1 in S. pombe caused no apparent effects on growth and division . However , mutation of a predicted regulatory site ( DB00156 -192 ) to aspartic acid resulted in hyperactivation of CMK1 activity in the presence of P62158 and causes cell cycle arrest in vivo . Arrest is also accompanied by morphological defects . These results suggest the presence of a P62158 -dependent protein kinase cascade in yeast and indicate that cmk1 may be important in cell cycle progression , a process known to be dependent on P62158 in eukaryotic cells . Early gastrointestinal regulatory peptide response to intestinal resection in the rat is stimulated by enteral glutamine supplementation . BACKGROUND : Intestinal resection stimulates the synthesis and release of gastrointestinal peptides that regulate the growth and adaptation of the mucosa . DB01174 nutrients are necessary for optimal proliferation and glutamine is the preferential nutrient to the small bowel . The interplay between glutamine and regulatory peptides could be important in treating short bowel syndrome . METHODS : 63 Sprague-Dawley rats were divided into 3 groups : resection ; transection , or controls . After intestinal resection animals were orally fed either a diet without glutamine or a glutamine-supplemented diet for 2 days . Transected animals and controls without prior surgery were fed the same two diets . Epidermal growth factor ( P01133 ) , transforming growth factor-alpha , insulin-like growth factors I and II ( P05019 and P01344 ) , peptide YY ( P10082 ) , and enteroglucagon were analyzed in mucosa from the proximal jejunum , distal ileum as well as in portal plasma when the animals were euthanized 72 h after surgery . RESULTS : Intestinal resection resulted in an early increase in portal plasma concentrations of P10082 , P01133 , enteroglucagon , and mucosal P01344 and P01133 content that were significant in glutamine-treated animals . Glutamine significantly increased P10082 in portal blood after resection ( p < 0.05 ) . CONCLUSION : Glutamine could be of importance for the functional adaptation of residual small bowel mucosa by increasing P10082 release . Effects of dutasteride on the expression of genes related to androgen metabolism and related pathway in human prostate cancer cell lines . Androgens play an important role in controlling the growth of the normal prostate gland and in the pathogenesis of benign prostate hyperplasia , and prostate cancer . Although testosterone is the main androgen secreted from the testes , dihydrotestosterone ( DB02901 ) , a more potent androgen converted from testosterone by 5alpha-reductase isozymes , type I and II , is the major androgen in the prostate cells . The aim of this study is to investigate the cellular and molecular effects of dutasteride , a potent inhibitor of 5alpha-reductase type I and type II , in androgen-responsive ( LNCaP ) and androgen-unresponsive ( DU145 ) human prostate cancer(PCa) cell lines . The expression pattern of 190 genes , selected on the basis of their proved or potential role in prostate cancerogenesis related to androgen signalling , were analysed using a low density home-made oligoarray ( AndroChip 2 ) . Our results show that dutasteride reduces cell viability and cell proliferation in both cell lines tested . AndroChip 2 gene signature identified in LNCaP a total of 11 genes differentially expressed ( FC > or= +/-1.5 ) . Eight of these genes , were overexpressed and three were underexpressed . Overexpressed genes included genes encoding for proteins involved in biosynthesis and metabolism of androgen ( P14061 ; P37058 ; P19099 ) , androgen receptor and androgen receptor co-regulators ( AR; P24385 ) , and signal transduction ( P04626 ; V- P62158 ; Q07889 ) whereas , underexpressed genes ( KLK3 ; P20151 ; Q15392 ) were androgen-regulated genes ( ARGs ) . No differentially expressed genes were scored in DU145 . Microarray data were confirmed by quantitative real-time PCR assay ( QRT-PCR ) . These data offer a selective genomic signature for dutasteride treatment in prostate epithelial cells and provide important insights in prostate cancer pathophysiology . An endothelial nitric oxide synthase inhibitor aggravates DB03429 -induced acute pancreatitis in rats . To clarify the role of nitric oxide ( NO ) in the development and progression of acute pancreatitis , we investigated the effect of different NO synthase inhibitors and NO donors on experimental pancreatitis in rats . Closed duodenal loop ( DB03429 ) -induced pancreatitis was produced in male Wistar rats , and the animals were treated with normal saline , the NO-synthase substrate L-arginine , the NO donor S-nitroso-N-acetylpenicillamine , aminoguanidine , which is a more powerful inhibitor of inducible NO synthase ( P35228 ) than is endothelial NO synthase ( P29474 ) , and N-nitro-L-arginine methyl ester ( L-NAME ) , a more powerful inhibitor of P29474 than of P35228 . All drugs were infused intravenously during a period of 6 or 12 h in each group . Pancreatic tissue was removed at 6 and 12 h after creating the DB03429 . L- DB00125 , S-nitroso-N-acetyl-penicillamine , and aminoguanidine treatment had no effect on the elevation of serum pancreatic enzymes , whereas L-NAME administration significantly exacerbated their elevation . Pathologically , L-NAME treatment resulted in a significantly worse histologic score at 6 and 12 h , especially in terms of the degree of hemorrhage , acinar cell necrosis , and microvascular thrombosis . Addition of L-arginine clearly reversed the effect of L-NAME . Neither the NO substrate nor NO donor could inhibit the progression of hemorrhagic pancreatitis in DB03429 -induced pancreatitis . DB02533 had no effect on the severity of the pancreatitis . We therefore concluded that NO production by P29474 may play a significant role in preventing the development and progression of acute pancreatitis . [ Proteolytic processing by dipeptidyl aminopeptidase IV generates receptor selectivity for peptide YY ( P10082 ) ] . Two receptor subtypes , Q03519 and P28062 , are known to mediate P10082 biological activity . P10082 1-36 binds to Q03519 and P28062 receptors with equal affinity , whereas the second endogenous form of P10082 , DB05004 , selectively binds to P28062 receptors . Dipeptidyl cleavage thus transforms an unselective Y agonist into a highly selective P28062 agonist , DB05004 . The enzyme responsible for this processing is unknown . Since P10082 has a proline in the penultimate position it is protected from the attack of most unspecific exopeptidases . Only a few exopeptidases are theoretically capable of generating DB05004 from P10082 1-36 . Of the enzymes tested only the dipeptidyl aminopeptidase IV ( P27487 , E.C. 3.4.14.5 ) cleaved DB00135 -Pro from P10082 1-36 with high activity . Since P27487 is found on the endothelial surface and brush border membranes it can be considered a candidate enzyme for generating DB05004 in vivo , thereby regulating the ratio of Q03519 / P49146 stimulation by P10082 . Emerging small molecule drugs . Dyslipidaemia is a major risk factor for cardiovascular diseases . Pharmacological lowering of LDL-C levels using statins reduces cardiovascular risk . However , a substantial residual risk persists especially in patients with type 2 diabetes mellitus . Because of the inverse association observed in epidemiological studies of HDL-C with the risk for cardiovascular diseases , novel therapeutic strategies to raise HDL-C levels or improve HDL functionality are developed as complementary therapy for cardiovascular diseases . However , until now most therapies targeting HDL-C levels failed in clinical trials because of side effects or absence of clinical benefits . This chapter will highlight the emerging small molecules currently developed and tested in clinical trials to pharmacologically modulate HDL-C and functionality including new P11597 inhibitors ( anacetrapib , evacetrapib ) , novel Q07869 agonists ( K-877 , CER-002 , P15924 -8658 , INT131 and DB05187 ) , LXR agonists ( ATI-111 , LXR-623 , XL-652 ) and RVX-208 . Treatment of Corneal Neovascularization Using Anti- P15692 DB00112 . Purpose. To evaluate antiangiogenic effect of local use of bevacizumab ( anti- P15692 antibody ) in patients with corneal neovascularization . Methods . Patients were divided into two groups . All patients suffered from some form of corneal neovascularization ( NV ) . Patients in group A received 0.2-0.5 mL of bevacizumab solution subconjunctivally ( concentration 25 mg/mL ) in a single dose . Group A included 28 eyes from 27 . Patients in group B applied bevacizumab eye drops twice daily ( concentration 2.5 mg/mL ) for two weeks . Group B included 38 eyes from 35 patients . We evaluated the number of corneal segments affected by NV , CDVA , and the incidence of complications and subjective complaints related to the treatment . The minimum follow-up period was six months . Results . By the 6-month follow-up , in group A the percentage reduction of the affected peripheral segments was 21.6 % and of the central segments was 9.6 % ; in group B the percentage reduction of the central segments was 22.7 % and of the central segments was 38.04 % . In both groups we noticed a statistically significant reduction in the extent of NV . Conclusion . The use of bevacizumab seems to be an effective and safe method in the treatment of corneal neovascularization , either in the subconjunctival or topical application form .

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interacts_with DB01182?

multiple_choice

Pathogenesis and treatment of thrombohemorrhagic diathesis in acute promyelocytic leukemia . Acute promyelocytic leukemia ( APL ) is a distinct subtype of myeloid leukemia characterized by t(15;17) chromosomal translocation , which involves the retinoic acid receptor-alpha ( P10276 ) . APL typically presents with a life-threatening hemorrhagic diathesis . Before the introduction of all-trans retinoic acid ( DB00755 ) for the cure of APL , fatal hemorrhages due , at least in part , to the APL-associated coagulopathy , were a major cause of induction remission failure . The laboratory abnormalities of blood coagulation found in these patients indicate the occurrence of a hypercoagulable state . Major determinants of the coagulopathy of APL are endogenous factors expressed by the leukemic cells , including procoagulant factors , fibrinolytic proteins , and non-specific proteolytic enzymes . In addition , these cells have an increased capacity to adhere to the vascular endothelium , and to secrete inflammatory cytokines [ i.e. interleukin-1beta ( IL-1beta ) and tumor necrosis factor ( P01375 ) ] , which in turn stimulate the expression of prothrombotic activities by endothelial cells and leukocytes . DB00755 can interfere with each of the principal hemostatic properties of the leukemic cell , thus reducing the APL cell procoagulant potential , in parallel to the induction of cellular differentiation . This effect occurs in vivo , in the bone marrow of APL patients receiving DB00755 , and is associated with the improvement of the bleeding symptoms . Therapy with arsenic trioxide ( ATO ) also beneficially affects coagulation in APL . However , early deaths from bleeding still remain a major problem in APL and further research is required in this field . In this review , we will summarize our current knowledge of the pathogenesis of the APL-associated coagulopathy and will overview the therapeutic approaches for the management of this complication . The low-potency , voltage-dependent Q12809 blocker propafenone -- molecular determinants and drug trapping . The molecular determinants of high-affinity human ether-a-go-go-related gene ( Q12809 ) potassium channel blockade by methanesulfonanilides include two aromatic residues ( Phe656 and Tyr652 ) on the inner helices ( S6 ) and residues on the pore helices that face into the inner cavity , but determinants for lower-affinity Q12809 blockers may be different . In this study , alanine-substituted Q12809 channel mutants of inner cavity residues were expressed in Xenopus laevis oocytes and were used to characterize the Q12809 channel binding site of the antiarrhythmic propafenone . DB01182 's blockade of Q12809 was strongly dependent on residue Phe656 but was insensitive or weakly sensitive to mutation of Tyr652 , Thr623 , Ser624 , Val625 , Gly648 , or Val659 and did not require functional inactivation . Homology models of Q12809 based on KcsA and MthK crystal structures , representing the closed and open forms of the channel , respectively , suggest propafenone is trapped in the inner cavity and is unable to interact exclusively with Phe656 in the closed state ( whereas exclusive interactions between propafenone and Phe656 are found in the open-channel model ) . These findings are supported by very slow recovery of wild-type Q12809 channels from block at -120 mV , but extremely rapid recovery of D540K channels that reopen at this potential . The experiments and modeling suggest that the open-state propafenone binding-site may be formed by the Phe656 residues alone . The binding site for propafenone ( which may involve pi-stacking interactions with two or more Phe656 side-chains ) is either perturbed or becomes less accessible because of closed-channel gating . This provides further evidence for the existence of gating-induced changes in the spatial location of Phe656 side chains . P08183 , Q9UNQ0 , and P60484 determine the response of glioblastoma to temozolomide and ABT-888 therapy . PURPOSE : Little is known about the optimal clinical use of ABT-888 ( veliparib ) for treatment of glioblastoma . ABT-888 is a PARP inhibitor undergoing extensive clinical evaluation in glioblastoma , because it may synergize with the standard-of-care temozolomide ( DB00853 ) . We have elucidated important factors controlling ABT-888 efficacy in glioblastoma . EXPERIMENTAL DESIGN : We used genetically engineered spontaneous glioblastoma mouse models and allograft models that were orthotopically transplanted into wild-type ( WT ) and Abcb1/Abcg2-deficient ( KO ) recipients . RESULTS : ABT-888/ DB00853 is not efficacious against p53;p16(Ink4a)/p19(Arf);K-Ras(v12);LucR allografts in wild-type recipients , indicating inherent resistance . Abcb1/Abcg2 mediated efflux of ABT-888 at the blood-brain barrier ( BBB ) causes a 5-fold reduction of ABT-888 brain penetration ( P < 0.0001 ) that was fully reversible by elacridar . Efficacy studies in WT and KO recipients and/or concomitant elacridar demonstrate that Abcb1/Abcg2 at the BBB and in tumor cells impair DB00853 /ABT-888 combination treatment efficacy . DB04881 also markedly improved DB00853 /ABT-888 combination treatment in the spontaneous p53;p16(Ink4a)/p19(Arf);K-Ras(v12);LucR glioblastoma model . Importantly , ABT-888 does enhance DB00853 efficacy in Pten deficient glioblastoma allografts and spontaneous tumors , even in Abcb1/Abcg2 proficient wild-type mice . Loss of P60484 occurs frequently in glioblastoma ( 36 % ) and in silico analysis on patient with glioblastoma samples revealed that it is associated with a worse overall survival ( 310 days vs. 620 days , n = 117 ) . CONCLUSIONS : The potential of ABT-888 in glioblastoma can best be demonstrated in patients with P60484 null tumors . Therefore , clinical trials with ABT-888 should evaluate these patients as a separate group . Importantly , inhibition of P08183 and Q9UNQ0 ( by elacridar ) may improve the efficacy of DB00853 /ABT-888 therapy in all glioblastoma patients . Targeted genome-wide methylation and gene expression analyses reveal signaling pathways involved in ovarian dysfunction after developmental EDC exposure in rats . Transient exposure to methoxychlor ( MXC ) , an environmental endocrine-disrupting chemical , during fetal and neonatal stages causes ovarian dysfunction in pubertal , adult , and aging animals . Adult animals have reduced number of ovulations and abnormal follicular composition associated with altered gene expression and DNA methylation patterns . To test the hypothesis that the ovarian epigenomic changes induced by MXC are detectable following the exposure period , leading to altered gene expression by adulthood , we conducted a targeted genome-wide methylation study using Nimblegen 3x720K CpG Island Plus RefSeq Promoter Arrays . Control ( vehicle ) , low-dose MXC ( 20 μg/kg/day ) , or high-dose MXC ( 100 mg/kg/day ) treatments were administered between Embryonic Day 19 and Postnatal Day ( P01160 ) 7 . Ovaries were collected at P01160 7 immediately after exposure or at adulthood , P01160 60 . Array hybridizations were conducted with genomic DNA after methylated DNA immunoprecipitation and the array data were analyzed . DNA methylation events were functionally annotated , and candidate loci common to all the treatments or unique to some treatments were identified . Specific loci encoding signaling molecules such as the regulatory subunit p85 of phosphoinositide-3-kinase , insulin-like growth factor-1 receptor , Harvey rat sarcoma viral oncogene , insulin receptor , and forkhead box protein O3 were identified to be hypermethylated in MXC-treated ovaries at P01160 7 and/or P01160 60 . Examination of gene expression changes with TaqMan low-density arrays revealed that nearly 25 % of the genes that were assayed were downregulated . These data demonstrate that key molecules in specific signaling pathways such as P60484 signaling , DB01277 signaling , or rapid estrogen signaling are epigenetically altered in MXC-exposed ovaries , which is associated with ovarian dysfunction and female infertility . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . The Antidepressant DB06594 Improves Memory Deterioration and Upregulates CREB and P23560 Gene Expression Levels in Unpredictable Chronic Mild Stress ( UCMS ) -Exposed Mice . DB06594 , a novel antidepressant with established clinical efficacy , acts as an agonist of melatonergic MT1 and P02795 receptors and as an antagonist of P28335 receptors . The present study was undertaken to investigate whether chronic treatment with agomelatine would block unpredictable chronic mild stress ( UCMS ) -induced cognitive deterioration in mice in passive avoidance ( PA ) , modified elevated plus maze ( mEPM ) , novel object recognition ( DB01434 ) , and Morris water maze ( MWM ) tests . Moreover , the effects of stress and agomelatine on brain-derived neurotrophic factor ( P23560 ) and cyclic adenosine monophosphate ( DB02527 ) response element binding protein ( CREB ) messenger ribonucleic acid ( mRNA ) levels in the hippocampus was also determined using quantitative real-time polymerase chain reaction ( RT-PCR ) . Male inbred BALB/c mice were treated with agomelatine ( 10 mg/kg , i.p. ) , melatonin ( 10 mg/kg ) , or vehicle daily for five weeks . The results of this study revealed that UCMS-exposed animals exhibited memory deterioration in the PA , mEPM , DB01434 , and MWM tests . The chronic administration of melatonin had a positive effect in the PA and +mEPM tests , whereas agomelatine had a partial effect . Both agomelatine and melatonin blocked stress-induced impairment in visual memory in the DB01434 test and reversed spatial learning and memory impairment in the stressed group in the MWM test . Quantitative RT-PCR revealed that CREB and P23560 gene expression levels were downregulated in UCMS-exposed mice , and these alterations were reversed by chronic agomelatine or melatonin treatment . Thus , agomelatine plays an important role in blocking stress-induced hippocampal memory deterioration and activates molecular mechanisms of memory storage in response to a learning experience . Botulinum neurotoxin A subtype 2 reduces pathological behaviors more effectively than subtype 1 in a rat Parkinson 's disease model . Recent reports indicate that interruption of acetylcholine release by intrastriatal injection of botulinum neurotoxin type A ( DB00083 ) in a rat Parkinson 's disease model reduces pathogenic behavior without adverse side effects such as memory dysfunction . Current knowledge suggests that DB00083 subtype 1 ( BoNT/A1 ) and DB00083 subtype 2 ( BoNT/A2 ) exert different effects . In the present study , we compared the effects of BoNT/A1 and BoNT/A2 on rotation behavior and in vivo cleavage of presynaptic protein P60880 in a rat unilateral 6-hydroxydopamine-induced Parkinson 's disease model . BoNT/A2 more effectively reduced pathogenic behavior by efficiently cleaving P60880 in the striatum compared with that of BoNT/A1 . Our results suggest that BoNT/A2 has greater clinical therapeutic value for treating subjects with Parkinson 's disease compared to that of BoNT/A1 . Cardiac channelopathies associated with infantile fatal ventricular arrhythmias : from the cradle to the bench . BACKGROUND : Fatal ventricular arrhythmias in the early period of life have been associated with cardiac channelopathies for decades , and postmortem analyses in P22304 victims have provided evidence of this association . However , the prevalence and functional properties of cardiac ion channel mutations in infantile fatal arrhythmia cases are not clear . METHODS AND RESULTS : Seven infants with potentially lethal arrhythmias at age < 1 year ( 5 males , age of onset 44.1 ± 72.1 days ) were genetically analyzed for P51787 , Q12809 , P15382 -5 , P63252 , Q14524 , P36382 , and P62158 by using denaturing high-performance liquid chromatography and direct sequencing . Whole-cell currents of wildtype and mutant channels were recorded and analyzed in Chinese hamster ovary cells transfected with Q14524 and Q12809 cDNA . In 5 of 7 patients , we identified 4 mutations ( p.N1774D , p.T290fsX53 , p.F1486del and p.N406K ) in Q14524 , and 1 mutation ( p.G628D ) in Q12809 . N1774D , F1486del , and N406K in Q14524 displayed tetrodotoxin-sensitive persistent late Na(+) currents . By contrast , Q14524 -T290fsX53 was nonfunctional . Q12809 -G628D exhibited loss of channel function . CONCLUSION : Genetic screening of 7 patients was used to demonstrate the high prevalence of cardiac channelopathies . Functional assays revealed both gain and loss of channel function in Q14524 mutations , as well as loss of function associated with the Q12809 mutation . Recent findings in the genetics of blood pressure and hypertension traits . We provide an overview of ongoing discovery efforts in the genetics of blood pressure ( BP ) and hypertension ( HTN ) traits . Two large genome-wide association meta-analyses of individuals of European descent were recently published , revealing ~13 new loci for BP traits . Only two of these loci harbor genes in a pathway known to affect BP ( P05093 and P01160 / P16860 ) . Functional variants in these loci are still unknown . Few genome-wide association studies ( GWAS ) of complex diseases have been published from non-European populations . The study of populations with different evolutionary history and linkage disequilibrium ( LD ) structure , such as individuals of African ancestry , may provide an opportunity to further narrow these regions to identify the causal gene(s) . Several collaborative efforts toward discovery of low-frequency variants and copy number variation for BP traits are currently underway . As evidence for new loci for complex diseases accumulates the assessment of the epidemiologic architecture of these variants in populations assumes higher priority . The impact of public health-relevant contexts such as diet , physical activity , psychosocial factors , and aging has not been examined for most common variants associated with BP . A common single nucleotide polymorphism can exacerbate long-QT type 2 syndrome leading to sudden infant death . BACKGROUND : Identification of infants at risk for sudden arrhythmic death remains one of the leading challenges of modern medicine . We present a family in which a common polymorphism ( single nucleotide polymorphism ) inherited from the father , combined with a stop codon mutation inherited from the mother ( both asymptomatic ) , led to 2 cases of sudden infant death . METHODS AND RESULTS : P51787 , Q12809 , Q14524 , P15382 , Q9Y6J6 , CACNA1c , CACNB2b , and P63252 genes were amplified and analyzed by direct sequencing . Functional electrophysiological studies were performed with the single nucleotide polymorphism and mutation expressed singly and in combination in Chinese ovary ( CHO- P04264 ) and COS-1 cells . An asymptomatic woman presenting after the death of her 2-day-old infant and spontaneous abortion of a second baby in the first trimester was referred for genetic analysis . The newborn infant had nearly incessant ventricular tachycardia while in utero and a prolonged QTc ( 560 ms ) . The mother was asymptomatic but displayed a prolonged QTc . Genetic screening of the mother revealed a heterozygous nonsense mutation ( P926AfsX14 ) in Q12809 , predicting a stop codon . The father was asymptomatic with a normal QTc but had a heterozygous polymorphism ( K897T ) in Q12809 . The baby who died at 2 days of age and the aborted fetus inherited both K897T and P926AfsX14 . Heterologous coexpression of K897T and P926AfsX14 led to loss of function of Q12809 current much greater than expression of K897T or P926AfsX14 alone . CONCLUSIONS : Our data suggest that a common polymorphism ( K897T ) can markedly accentuate the loss of function of mildly defective Q12809 channels , leading to long-QT syndrome-mediated arrhythmias and sudden infant death . ICE/ P29466 inhibitors as novel anti-inflammatory drugs . In recent years , several strategies that selectively inhibit pro-inflammatory cytokines , have yielded effective protein-based therapies for inflammatory disorders , validating the therapeutic hypothesis that intervention in cytokine signalling can provide clinical benefit . However , these protein-based products must be administered by injection , a constraint associated with inconvenience , adverse effects and expense for patients , caregivers and insurers . Besides interfering with the effects of cytokines such as P01375 or IL-1beta that have already been produced , inhibition of pro-inflammatory cytokine production or signalling with low-molecular weight orally-active drugs would combine the convenience of conventional pharmaceuticals with the focused efficacy of the protein therapies . Reducing IL-1beta and Q14116 production by inhibition of IL-1beta converting enzyme ( ICE , caspase-1 ) is one promising strategy because of the key roles of these cytokines in many inflammatory diseases . DB04875 , the first orally available , potent and selective ICE inhibitor to enter clinical trials , is currently under investigation in rheumatoid arthritis . DB01184 treatment for gastroparesis : demographic and pharmacogenetic characterization of clinical efficacy and side-effects . BACKGROUND : DB01184 is a useful alternative to metoclopramide for treatment of gastroparesis due to better tolerability . Effectiveness and side-effects from domperidone may be influenced by patient-related factors including polymorphisms in genes encoding drug-metabolizing enzymes , drug transporters , and domperidone targets . AIMS : The aim of this study was to determine if demographic and pharmacogenetic parameters of patients receiving domperidone are associated with response to treatment or side-effects . METHODS : Patients treated with domperidone for gastroparesis provided saliva samples from which DNA was extracted . Fourteen single-nucleotide polymorphisms ( SNPs ) in seven candidate genes ( P08183 , P10635 , P14416 , P15382 , Q9Y6J6 , Q12809 , P51787 ) were used for genotyping . SNP microarrays were used to assess single-nucleotide polymorphisms in the ADRA1A , P35368 , and P25100 loci . RESULTS : Forty-eight patients treated with domperidone participated in the study . DNA was successfully obtained from each patient . Age was associated with effectiveness of domperidone ( p=0.0088 ) . Genetic polymorphism in Q12809 was associated with effectiveness of domperidone ( p=0.041 ) . The efficacious dose was associated with polymorphism in P08183 gene ( p=0.0277 ) . The side-effects of domperidone were significantly associated with the SNPs in the promoter region of P25100 gene . CONCLUSIONS : Genetic characteristics associated with response to domperidone therapy included polymorphisms in the drug transporter gene P08183 , the potassium channel Q12809 gene , and α1D -- adrenoceptor P25100 gene . Age was associated with a beneficial response to domperidone . If verified in a larger population , this information might be used to help determine which patients with gastroparesis might respond to domperidone and avoid treatment in those who might develop side-effects . Altered gene expression by low-dose arsenic exposure in humans and cultured cardiomyocytes : assessment by real-time PCR arrays . Chronic arsenic exposure results in higher risk of skin , lung , and bladder cancer , as well as cardiovascular disease and diabetes . The purpose of this study was to investigate the effects on expression of selected genes in the blood lymphocytes from 159 people exposed chronically to arsenic in their drinking water using a novel RT-PCR TaqMan low-density array ( TLDA ) . We found that expression of tumor necrosis factor-α ( P01375 -α ) , which activates both inflammation and NF-κB-dependent survival pathways , was strongly associated with water and urinary arsenic levels . Expression of P22460 , which encodes a potassium ion channel protein , was positively associated with water and toe nail arsenic levels . Expression of 2 and 11 genes were positively associated with nail and urinary arsenic , respectively . Because arsenic exposure has been reported to be associated with long QT intervals and vascular disease in humans , we also used this TLDA for analysis of gene expression in human cardiomyocytes exposed to arsenic in vitro . Expression of the ion-channel genes CACNA1 , Q12809 , P51787 and P15382 were down-regulated by 1-μM arsenic . Alteration of some common pathways , including those involved in oxidative stress , inflammatory signaling , and ion-channel function , may underlay the seemingly disparate array of arsenic-associated diseases , such as cancer , cardiovascular disease , and diabetes . Genomic structure and chromosome location of the murine Q01064 phosphodiesterase gene . Cyclic nucleotide phosphodiesterases ( PDEs ) catalyze the hydrolysis of DB02527 and cGMP , thereby participating in regulation of the intracellular concentrations of these second messengers . The PDE1 family is defined by regulation of activity by calcium and calmodulin . We have cloned and characterized the mouse Q01064 gene , which encodes the 63-kDa calcium/calmodulin-dependent PDE ( P62158 -PDE ) , an isozyme that is expressed in the CNS in the olfactory tract , dentate gyrus , and striatum and may participate in learning , memory , and regulation of phosphorylation of Q9UD71 in dopaminergic neurons . We screened an I-129/SvJ mouse genomic library and identified exons 2-13 of the Q01064 gene that span 8.4 kb of genomic DNA . Exons range from 67 to 205 nucleotides and introns from 91 to 2250 nucleotides in length . Exon 1 was not present in the 3 kb of genomic DNA 5' to exon 2 in our clones . The mouse Q01064 gene shares many similar or identical exon boundaries as well as considerable sequence identity with the rat Q07343 and Q08499 genes and the Drosophila dunce DB02527 -specific PDE gene dnc , suggesting that these genes all arose from a common ancestor . Using fluorescence in situ hybridization , we localized the Q01064 gene to the distal tip of mouse Chromosome ( Chr ) 15 . DB00115 responsive homocystinuria and megaloblastic anemia : heterogeneity in methylcobalamin deficiency . A male infant with methyl-B12 deficiency ( cblE ) presented at age 6 weeks with lethargy , staring spells , and vomiting . He later became hypotonic and unresponsive to stimuli and required intubation and ventilation . He had homocystinuria and hypomethioninemia with megaloblastic anemia but normal serum folate and vitamin B12 concentrations . No methylmalonic aciduria was detected . Fibroblasts , cultured from the patient , were unable to grow in medium in which homocysteine replaced methionine and incorporated abnormally small amounts of [ 14C ] -methyl- DB00116 but normal amounts of [ 14C ] -propionate into protein . Methyl-B12 content of fibroblasts was low , while the adenosyl-B12 content was normal . Q99707 activity was decreased when the assay was performed under both optimal and suboptimal reducing conditions , suggesting heterogeneity in the cblE disease . The patient responded dramatically to hydroxocobalamin treatment . Homocystinuria disappeared after 10 days of therapy , and methionine was normalized after 3 weeks . Psychometric testing at age 15 months showed a developmental age of 9 months . Design and synthesis of 3,5-disubstituted-1,2,4-oxadiazoles as potent inhibitors of phosphodiesterase4b2 . A series of 3,5-disubstituted-1,2,4-oxadiazoles has been prepared and evaluated for phosphodiesterase inhibition ( PDE4B2 ) . Among the prepared 3,5-disubstituted-1,2,4-oxadiazoles , compound 9a is the most potent inhibitor ( PDE4B2 IC(50) = 5.28 μm ) . Structure-activity relationship studies of 3,5-disubstituted-1,2,4-oxadiazoles revealed that substituents 3-cyclopentyloxy-4-methoxyphenyl group at 3-position and cyclic ring bearing heteroatoms at 5-position are important for activity . Molecular modeling study of the 3,5-disubstituted-1,2,4-oxadiazoles with Q07343 has shown similar interactions of 3-cyclopentyloxy-4-methoxyphenyl group ; however , heteroatom ring is slightly deviating when compared to DB01791 . 3-(3-Cyclopentyloxy-4-methoxyphenyl)-5-(piperidin-4-yl)-1,2,4-oxadiazole ( 9a ) exhibited good analgesic and antiinflammatory activities in formalin-induced pain in mice and carrageenan-induced paw edema model in rat . Polyamines and membrane transporters . In recent years , our understanding of the importance of membrane transporters ( MTs ) in the disposition of and response to drugs has increased significantly . MTs are proteins that regulate the transport of endogenous molecules and xenobiotics across the cell membrane . In mammals , two super-families have been identified : DB00171 -binding cassette ( DB01048 ) and solute carrier ( O00585 ) transporters . There is evidence that MTs might mediate polyamines ( PA ) transport . PA are ubiquitous polycations which are found in all living cells . In mammalian cells , three major PA are synthesised : putrescine , spermidine and spermine ; whilst the decarboxylated arginine ( agmatine ) is not produced by mammals but is synthesised by plants and bacteria . In addition , research in the PA field suggests that PA are transported into cells via a specific transporter , the polyamine transport system(s) ( Q03393 ) . Although the Q03393 has not been fully defined , there is evidence that some of the known MTs might be involved in PA transport . In this mini review , eight O00585 transporters will be reviewed and their potential to mediate PA transport in human cells discussed . These transporters are O15245 , O15244 , O75751 , Q96FL8 , P30825 , P08195 , SLC12A8A , and Q86VW1 . Preliminary data from our laboratory have revealed that O15245 might be involved in the PA uptake ; in addition to one member of ABC superfamily ( P08183 protein ) might also mediate the efflux of polyamine like molecules . Regulation of hepatic lecithin:retinol acyltransferase activity by retinoic acid receptor-selective retinoids . The microsomal enzyme O95237 esterifies retinol and has been implicated in the hepatic storage of vitamin A . Previously , we showed that hepatic O95237 activity is negligible during vitamin A deficiency and that all-trans-retinoic acid ( all-trans-RA ) rapidly induces the activity of liver O95237 in retinoid-deficient rats . In the present studies , we have examined the ability of natural and synthetic retinoids to induce liver O95237 activity in retinoid-deficient rats . The natural retinoids retinol , all-trans-RA ( 100 microg ) , 9- DB00982 , or equal molar amounts of other retinoids were injected ip and O95237 specific activity was measured in liver homogenates 17-18 h later . In retinoid-deficient rats , liver O95237 activity was extremely low [ 0.13 +/- 0.03 pmol retinyl ester ( RE ) /min/mg liver protein , mean +/- SE ] . The natural retinoids retinol and all-trans-RA strongly induced O95237 activity ( 12.71 +/- 1.09 and 13.10 +/- 1.55 pmol RE/min/mg , respectively ) , whereas 9- DB00982 induced a lower level of O95237 activity ( 3.96 +/- 1.88 pmol RE/min/mg , P < 0.001 vs all-trans-RA ) . The retinoic acid receptor ( RAR ) -selective analog ( RAR pan-agonist ) all-trans-UAB8 and the P10276 -selective retinoid Am580 also strongly induced O95237 activity . In contrast , neither RXR-selective agonists nor retinoids having a retro structure were active . For retinoids with significant P10276 binding activity there was a strong direct correlation between receptor binding in vitro and the ability to induce hepatic O95237 activity in vivo ( r2 = 0.920 ) . These data implicate the RARs in the induction of hepatic O95237 and suggest a predominant role for P10276 -active ligands . Long-lasting effects of elevated neonatal leptin on rat hippocampal function , synaptic proteins and DB01221 receptor subunits . The high circulating levels of leptin in neonatal rodents do not seem to be regulating energy balance at this age , but rather may play an important role for brain development . We tested the hypothesis that high neonatal leptin levels modify hippocampal function and production of synaptic proteins with possible long-term consequences on long-term potentiation ( LTP ) in adulthood . We first showed that in postnatal day ( P01160 ) 10 neonates , acute leptin treatment functionally activated leptin receptors ( ObR ) in the P00915 and DG regions of the hippocampus through the induction of phosphoERK1/2 , but not phosphoSTAT3 protein although both phospho-proteins were induced in the arcuate nucleus . We next examined whether chronic leptin administration ( 3 mg/kg BW , intraperitoneally ) during the first 2 weeks of life ( postnatal day , P01160 2-14 ) produces a functional signal in the hippocampus that alters the expression of DB01221 receptor subunits ( Q9UHB4 , Q12879 , Q13224 ) , synaptic proteins and LTP in the short and long-term . In P01160 10 as in adults ( P01160 70 ) rats , chronic leptin treatment increased Q9UHB4 expression in the hippocampus while reducing Q13224 protein levels . Elevated hippocampal concentrations of synapsin2A and synaptophysin were detected during leptin treatment on P01160 10 suggesting increased neurotransmitter release . In adults , only P60880 expression was increased after neonatal leptin treatment . LTP was reduced dramatically by leptin treatment in preweaning rats although the changes did not persist until adulthood . Elevated exposure to leptin during a critical period of neonatal hippocampal development might serve to enhance DB01221 -dependent functions other than LTP and have important effects on synaptogenesis and neurotransmitter release . Enhancement of L-cystine transport activity and its relation to Q9UPY5 gene induction at the blood-brain barrier by diethyl maleate treatment . The purpose of the present study was to elucidate the mechanism of enhancement of L-cystine uptake at the blood-brain barrier ( BBB ) . The uptake of [(14)C]L-cystine and [(3)H]L-glutamic acid ( L- DB00142 ) was determined using a mouse brain endothelial cell line ( MBEC4 ) as an in vitro BBB model . The mRNA levels of L-cystine/L- DB00142 exchanger , system x(c)(-) , which consists of Q9UPY5 and P08195 , were determined by quantitative real-time reverse transcription-polymerase chain reaction analysis . The [(14)C]L-cystine uptake by MBEC4 cells appeared to be mediated via an Na(+)-independent saturable process . The corresponding Michaelis-Menten constant ( K(m) ) was 63.7 microM . In the presence of L- DB00142 , there was competitive inhibition with an inhibition constant ( K(i) ) of 83.5 microM . [(3)H]L- DB00142 uptake in the absence of Na(+) was saturable with a K(m) of 48.1 microM , and it exhibited competitive inhibition with a K(i) of 24.9 microM in the presence of L-cystine . The mutual inhibition between L-cystine and L- DB00142 and the type of inhibition suggest that system x(c)(-) operates in MBEC4 cells . The Q9UPY5 and P08195 mRNAs were expressed in MBEC4 cells and , following diethyl maleate ( DEM ) treatment , the Q9UPY5 mRNA level and L-cystine uptake in MBEC4 cells were enhanced in parallel with an increase in DEM concentration ( up to 500 microM ) . Concomitantly , the glutathione concentration in MBEC4 cells was increased . In conclusion , system x(c)(-)-mediated L-cystine uptake takes place in MBEC4 cells . DB00138 transport via system x(c)(-) at the BBB is likely to be induced under oxidative stress conditions following DEM treatment due to enhanced transcription of the Q9UPY5 gene . [ DB05812 acetate : a novel therapeutic option in hormone-refractory prostate cancer ] . Until recently , only therapy with docetaxel and prednisone has been shown to prolong survival in men with hormonorefractory metastatic prostate cancer . With approvals of sipuleucel-T , cabazitaxel , and abiraterone acetate , all based on improvement in overall survival , the scenary for management of men with metastatic prostate cancer has dramatically changed . DB05812 acetate was developed to specifically inhibit cytochrome P450 (CYP)17A1 , which is an essential enzyme in the biosynthesis of testosterone . In the phase III , the trial treatment with abiraterone acetate plus prednisone prolongs overall survival relative to prednisone alone in patients with metastatic castration-resistant prostate cancer who have disease progression after treatment with docetaxel and associated with an acceptable tolerability profile , which was generally similar to that of the placebo plus prednisone group . However , adverse events resulting from elevated mineralocorticoid levels because of P05093 inhibition , fluid retention and oedema , hypokalaemia , hypertension occurred in significantly more in abiraterone acetate plus prednisone than in placebo plus prednisone . DB09337 cleaning in P01160 experiments at the ETR and Q99707 . Functional selectivity of hallucinogenic phenethylamine and phenylisopropylamine derivatives at human 5-hydroxytryptamine (5-HT)2A and P28335 receptors . 2,5-Dimethoxy-4-substituted phenylisopropylamines and phenethylamines are 5-hydroxytryptamine ( serotonin ) ( 5-HT ) (2A/2C) agonists . The former are partial to full agonists , whereas the latter are partial to weak agonists . However , most data come from studies analyzing phospholipase C ( P98160 ) -mediated responses , although additional effectors [ e.g. , phospholipase A(2) ( PLA(2) ) ] are associated with these receptors . We compared two homologous series of phenylisopropylamines and phenethylamines measuring both PLA(2) and P98160 responses in Chinese hamster ovary- P04264 cells expressing human 5-HT(2A) or 5-HT(2C) receptors . In addition , we assayed both groups of compounds as head shake inducers in rats . At the 5-HT(2C) receptor , most compounds were partial agonists for both pathways . Relative efficacy of some phenylisopropylamines was higher for both responses compared with their phenethylamine counterparts , whereas for others , no differences were found . At the 5-HT(2A) receptor , most compounds behaved as partial agonists , but unlike findings at 5-HT(2C) receptors , all phenylisopropylamines were more efficacious than their phenethylamine counterparts . 2,5-Dimethoxyphenylisopropylamine activated only the P98160 pathway at both receptor subtypes , 2,5-dimethoxyphenethylamine was selective for P98160 at the 5-HT(2C) receptor , and 2,5-dimethoxy-4-nitrophenethylamine was PLA(2)-specific at the 5-HT(2A) receptor . For both receptors , the rank order of efficacy of compounds differed depending upon which response was measured . The phenylisopropylamines were strong head shake inducers , whereas their phenethylamine congeners were not , in agreement with in vitro results and the involvement of 5-HT(2A) receptors in the head shake response . Our results support the concept of functional selectivity and indicate that subtle changes in ligand structure can result in significant differences in the cellular signaling profile .

MH_train_1355

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interacts_with DB00338?

multiple_choice

DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . FcεRI stimulation promotes the differentiation of histamine receptor 1-expressing inflammatory macrophages . BACKGROUND : Monocyte differentiation into dendritic cells or macrophages and recruitment to peripheral organs in chronic inflammatory diseases are directed by allergen challenge via FcεRI as well as the nature of soluble factors in the microenvironment . High-affinity receptor for IgE stimulation of effector cells results in the release of histamine , which acts on various histamine receptors ( HR ) 1-4 , expressed by immune cells . METHODS : We examined the effect of FcεRI stimulation of human monocytes on P35367 expression and function of differentiating cells . The mRNA levels of P35367 , P25021 and histidine decarboxylase of differentiating cells were detected by quantitative real-time PCR . Expression of CD1c , CD11c , P34810 and Q86VB7 was detected by flow cytometry . Amount of histamine , P05231 and IL-12p70 in the cell culture was measured with the help of cytometric bead arrays or ELISA assays . Numbers of P35367 -expressing macrophages were evaluated by immunofluorescence double staining of P34810 and P35367 on human skin sections . RESULTS : We demonstrated that FcεRI stimulation promotes the generation of P35367 -expressing macrophage-like cells with enhanced histamine biosynthesis and P35367 -mediated proinflammatory properties . Supporting our in vitro findings , high numbers of P35367 -expressing P34810 (pos) macrophages were detected in the dermis of atopic dermatitis ( AD ) skin lesions . CONCLUSION : Our observations point to a close histamine-/HR-mediated activation of dermal macrophages , leading to modified cell differentiation and responsiveness via P35367 , which might contribute to the aggravation of allergic skin inflammation in AD . Q9Y6X2 negatively regulates O14788 -mediated osteoclastogenesis directly in osteoclast precursors and indirectly via osteoblasts . Cytokine signaling via various transcription factors regulates receptor activator of nuclear factor ( NF ) -kappaB ligand ( O14788 ) -mediated osteoclast differentiation from monocyte/macrophage lineage cells involved in propagation and resolution of inflammatory bone destruction . Protein inhibitor of activated P40763 ( Q9Y6X2 ) was initially identified as a molecule that inhibits DNA binding of P40763 and regulates many transcription factors through distinct mechanisms . To analyze Q9Y6X2 function in osteoclasts in vivo , we have generated transgenic mice in which Q9Y6X2 is specifically expressed in the osteoclast lineage using the tartrate-resistant acid phosphatase ( TRAP ) gene promoter . Q9Y6X2 transgenic mice showed an osteopetrotic phenotype due to impairment of osteoclast differentiation . Overexpression of Q9Y6X2 in RAW264.7 cells suppressed O14788 -induced osteoclastogenesis by inhibiting the expression of c-Fos and O95644 . Interestingly , Q9Y6X2 inhibits the transcriptional activity of microphthalmia-associated transcription factor ( O75030 ) independent of sumoylation . Down-regulation of Q9Y6X2 markedly enhances O14788 -mediated osteoclastogenesis in RAW264.7 cells . Furthermore , overexpression of Q9Y6X2 in mouse primary osteoblast ( DB00925 ) , down-regulates O14788 expression induced by interleukin-6 ( P05231 ) cytokine family , and inhibits osteoclast formation from bone marrow macrophages ( BMMs ) in vitro coculture system . Down-regulation of Q9Y6X2 leads to the accelerated expression of O14788 in DB00925 stimulated with P05231 and soluble P05231 receptor ( sIL-6R ) . Taken together , our results clearly indicate that Q9Y6X2 negatively regulates O14788 -mediated osteoclastogenesis directly in osteoclast precursors and indirectly via osteoblasts . Genotype frequencies of 50 polymorphisms for 241 Japanese non-cancer patients . This paper lists the genotype frequencies of 50 polymorphisms of 37 genes ( P05091 , P07550 , P13945 , P21964 , P16671 , P25025 , P24385 , P35354 , P11509 , P05093 , P11511 , IGF1 , IL-1A , IL-1B , IL-1RN , IL-1R1 , P05231 , P10145 , P22301 , P41159 , Le , L-myc , P05164 , Q99707 , P42898 , P21397 , P15559 , O15527 , p53 , p73 , Se , P31213 , TGF-B , P01375 -A , P01375 -B , P18074 , and P18887 ) and 6 sets of combined genotype frequencies for 241 non-cancer Japanese outpatients . Though the genotype frequencies of 25 polymorphisms have already been reported in our previous papers , 15 polymorphisms ( P16671 A52C , P25025 C785T , P24385 G870A , IGF1 C/T at intron 2 and G2502T , IL-1A 46-bp VNTR , IL-1R1 C-116T , P05231 Ins/Del 17C , P10145 A-278T and C74T , IL- 10 T-819C , P41159 A-2548G , P31213 2-bp VNTR , P18074 Lys751Gln , and P18887 Arg399Gln ) and six sets of combined genotype frequencies ( IL-1B C-31T and IL-1A C-889T , IL-1B C-31T and IL-1RN 86-bp VNTR , IL-1B C-31T and IL-1R1 C-116T , P01375 -A G-308A and P01375 -B A252G , P31213 Val89Leu and 2-bp VNTR , and P18887 Arg399Gln and P18074 Lys751Gln ) were reported in this paper for the first time for Japanese . Although microarray technology will produce this kind of information in near future , this is the first document that reports the genotype/allele frequencies among Japanese for an archival purpose . Dimerization effect of sucrose octasulfate on rat P05230 . Fibroblast growth factors ( FGFs ) constitute a family of at least 23 structurally related heparin-binding proteins that are involved in regulation of cell growth , survival , differentiation and migration . DB01901 ( SOS ) , a chemical analogue of heparin , has been demonstrated to activate FGF signalling pathways . The structure of rat P05230 crystallized in the presence of SOS has been determined at 2.2 A resolution . SOS-mediated dimerization of P05230 was observed , which was further supported by gel-filtration experiments . The major contributors to the sulfate-binding sites in rat P05230 are Lys113 , Lys118 , Arg122 and Lys128 . An arginine at position 116 is a consensus residue in mammalian FGF molecules ; however , it is a serine in rat P05230 . This difference may be important for SOS-mediated P05230 dimerization in rat . Molecular markers of programmed cell death in donor hearts before transplantation . BACKGROUND : In this study we investigate whether pro-apoptotic , pro-inflammatory and other early signaling markers indicative of increased propensity for cell death processes were evident in human donor heart allografts immediately before transplantation , and whether there is an association with primary graft failure . METHODS : A prospective study was performed utilizing donor left atrial myocardium collected at the time of implantation of hearts from brain-dead donors ( BDD , n = 29 ) . In addition , to explore the potential of donor hearts from donation after circulatory death ( P81605 ) , myocardial samples were obtained during transplantation of lungs from P81605 donors ( n = 6 ) . A comparator reference group ( n = 7 ) consisted of left atrial specimens from patients undergoing mitral valve surgery . RESULTS : Significantly raised levels of caspase-3 specific activity , activated hypoxia inducible factor-1 ( HIF-1α ) and 8-hydroxy-2'-deoxyguanosine were evident in the transplanted hearts ( from BDD ) that developed primary graft failure ( n = 11 ) . P81605 hearts did not differ from BDD with regard to mRNA expression levels of FAS , Bax , P05231 and caspase-3 . Although P81605 hearts exhibited lower caspase-3 specific activity and activated hypoxia-inducible factor-1 protein , they had higher levels of mRNA for NF-κB , Bnip3 and caspase-1 mRNA . Increased 8-hydroxy-2'-deoxyguanosine levels reflected greater oxidative stress and reactive oxygen species-related DNA fragmentation . CONCLUSIONS : Our data indicate a significant role of pro-apoptotic and pro-inflammatory activity in allografts that subsequently exhibit primary graft failure . The relatively lower levels of apoptotic and inflammatory activity in P81605 hearts suggest they may represent a potentially usable donor cardiac allograft pool . This possibility requires further detailed molecular and clinical research . Guggulsterone inhibits angiogenesis by blocking P40763 and P15692 expression in colon cancer cells . The plant sterol guggulsterone has been shown to exert anti-tumor effects , making it a candidate chemotherapeutic agent . We investigated the anti-tumor effects of guggulsterone on colon cancer cells and elucidated the underlying molecular mechanisms related to angiogenesis . The apoptotic effects of guggulsterone were examined by cell survival assay . Western blot analysis was used to determine the levels of various down-stream intracellular proteins involved in angiogenesis , including signal transducer and activator of transcription 3 ( P40763 ) , vascular endothelial growth factor ( P15692 ) , hypoxia-inducible factor-1alpha ( HIF-1alpha ) and aryl hydrocarbon receptor nuclear translocator ( P27540 ) . Using chromatin immunoprecipitation assay , we tested whether guggulsterone affects the recruitment of P40763 , P27540 and HIF-1alpha to the human P15692 promoter . To investigate the effect of guggulsterone on vascular endothelial cell migration and invasion , tube formation and migration assays were conducted using human umbilical vein endothelial cells ( HUVECs ) . Matrix metalloproteinase ( MMP ) -2 and -9 activities were measured by gelatin zymography . Guggulsterone significantly reduced cell viability in colon cancer cells in a dose-dependent manner and blocked P15692 , P27540 and P40763 expression prominently in hypoxic conditions . The recruitment of P40763 and P27540 , but not HIF-1alpha , to the P15692 promoter was inhibited by guggulsterone treatment . HUVECs produced much foreshortened and severely broken tubes and showed decreased migration activity under guggulsterone effects . In addition , zymography revealed that P08253 and -9 enzyme activities were markedly lower in the presence of guggulsterone . The results of this study suggest that guggulsterone not only induces apoptosis , but also inhibits angiogenesis and metastasis in colon cancer cells by blocking P40763 and P15692 expression , suggesting its therapeutic potential in the treatment of colorectal cancer . Increases in stimulated secretion of proinflammatory cytokines by blood monocytes following arousal of negative affect : the role of insulin resistance as moderator . We examined the effect of negative affect on changes in stimulated secretion of cytokines by blood monocytes and determined whether insulin resistance ( IR ) , as indexed by the Homeostasis Model Assessment ( HOMA ) , moderated these associations in 58 healthy men ( aged 18-65 years ) . Blood samples and ratings of negative affect were collected at rest and 15min following subjects ' participation in the Anger Recall Interview ( Q9Y4X5 ) . Assessment of lipopolysaccharide ( LPS ) -stimulated secretion of IL-1beta , P05231 , and P01375 was accomplished by ELISA of supernatant . Regression models controlling for age , body mass index , and race/ethnicity revealed that higher HOMA-IR values were associated with larger stress-induced increases in IL-1beta and P01375 ( p < .05 ) . Furthermore , arousal of negative affect during the Q9Y4X5 was differentially associated with stress-induced changes in stimulated secretion of P01375 and P05231 as a function of HOMA-IR ( p < .05 ) . Increases in stimulated cytokine secretion were associated with arousal of negative affect , but only among men with higher HOMA-IR values . Among men with lower HOMA-IR values , arousal of negative affect was associated with diminished cytokine secretion . Collectively , these data suggest that the HOMA-IR moderates the impact that arousal of negative affect has on the ability of blood monocytes to secrete inflammatory cytokines in response to LPS . Stress-induced increases in cytokine secretion among high HOMA-IR men are consistent with the role of inflammation in cardiovascular disease , hypertension , type 2 diabetes as well as the metabolic syndrome and underscore the relevance of negative affect in the etiology of these inflammatory conditions . DB06698 treatment in managing vertigo and improving vestibular compensation : clarification . DB06698 dihydrochloride ( betahistine ) is currently used in the management of vertigo and vestibular pathologies with different aetiologies . The main goal of this review is to clarify the mechanisms of action of this drug , responsible for the symptomatic relief of vertigo and the improvement of vestibular compensation . The review starts with a brief summary recalling the role of histamine as a neuromodulator/neurotransmitter in the control of the vestibular functions , and the role of the histaminergic system in vestibular compensation . Then are presented data recorded in animal models demonstrating that betahistine efficacy can be explained by mechanisms targeting the histamine receptors ( HRs ) at three different levels : the vascular tree , with an increase of cochlear and vestibular blood flow involving the P35367 ; the central nervous system , with an increase of histamine turnover implicating the Q9Y5N1 , and the peripheral labyrinth , with a decrease of vestibular input implying the Q9Y5N1 / Q9H3N8 . Clinical data from vestibular loss patients show the impact of betahistine treatment for the long-term control of vertigo , improvement of balance and quality of life that can be explained by these mechanisms of action . However , two conditions , at least , are required for reaching the betahistine therapeutic effect : the dose and the duration of treatment . Experimental and clinical data supporting these requirements are exposed in the last part of this review . Extensive crosslinking of P20273 by epratuzumab triggers P11274 signaling and caspase-dependent apoptosis in human lymphoma cells . DB04958 has demonstrated therapeutic activity in patients with non-Hodgkin lymphoma , acute lymphoblastic leukemia , systemic lupus erythematosus , and Sjögren 's syndrome , but its mechanism of affecting normal and malignant B cells remains incompletely understood . We reported previously that epratuzumab displayed in vitro cytotoxicity to P20273 -expressing Burkitt lymphoma cell lines ( Daudi and Ramos ) only when immobilized on plates or combined with a crosslinking antibody plus a suboptimal amount of anti-IgM ( 1 μg/mL ) . Herein , we show that , in the absence of additional anti-IgM ligation , extensive crosslinking of P20273 by plate-immobilized epratuzumab induced intracellular changes in Daudi cells similar to ligating B-cell antigen receptor with a sufficiently high amount of anti-IgM ( 10 μg/mL ) . Specifically , either treatment led to phosphorylation of P20273 , CD79a and CD79b , along with their translocation to lipid rafts , both of which were essential for effecting caspase-dependent apoptosis . Moreover , such immobilization induced stabilization of F-actin , phosphorylation of Lyn , ERKs and JNKs , generation of reactive oxygen species ( ROS ) , decrease in mitochondria membrane potential ( Δψm ) , upregulation of pro-apoptotic Bax , and downregulation of anti-apoptotic Bcl-xl and Mcl-1 . The physiological relevance of immobilized epratuzumab was implicated by noting that several of its in vitro effects , including apoptosis , drop in Δψm , and generation of ROS , could be observed with soluble epratuzumab in Daudi cells co-cultivated with human umbilical vein endothelial cells . These results suggest that the in vivo mechanism of non-ligand-blocking epratuzumab may , in part , involve the unmasking of P20273 to facilitate the trans-interaction of B cells with vascular endothelium . Mechanism of copper transport at the blood-cerebrospinal fluid barrier : influence of iron deficiency in an in vitro model . DB09130 ( Cu ) is an essential trace element that requires tight homeostatic regulation to ensure appropriate supply while not causing cytotoxicity due to its strong redox potential . Our previous in vivo study has shown that iron deficiency ( FeD ) increases Cu levels in brain tissues , particularly in the choroid plexus , where the blood-cerebrospinal fluid ( P04141 ) barrier resides . This study was designed to elucidate the mechanism by which FeD results in excess Cu accumulation at the blood- P04141 barrier . The effect of FeD on cellular Cu retention and transporters Cu transporter-1 ( Ctr1 ) , divalent metal transporter 1 ( DMT1 ) , antioxidant protein-1 ( O00244 ) and Q04656 was examined in choroidal epithelial Z310 cells . The results revealed that deferoximine treatment ( FeD ) resulted in 70 % increase in cellular Cu retention ( P < 0.05 ) . A significant increase in the mRNA levels of DMT1 , but not Ctr1 , was also observed after FeD treatment , suggesting a critical role of DMT1 in cellular Cu regulation during FeD . Knocking down Ctr1 or DMT1 resulted in significantly lower Cu uptake by Z310 cells , whereas the knocking down of O00244 or Q04656 led to substantial increases of cellular retention of Cu . Taken together , these results suggest that Ctr1 , DMT1 , O00244 and Q04656 contribute to Cu transport at the blood- P04141 barrier , and that the accumulation of intracellular Cu found in the Z310 cells during FeD appears to be mediated , at least in part , via the upregulation of DMT1 after FeD treatment . Autocrine/Paracrine secretion of P05231 family cytokines causes angiotensin II-induced delayed P40763 activation . We recently reported that angiotensin II ( AngII ) biphasically activates the JAK/ P35610 pathway and induces delayed phosphorylation of P40763 in the late stage ( 120 min ) in cardiomyocytes . This study was designed to determine the mechanism of delayed phosphorylation of P40763 . Conditioned medium prepared from AngII-stimulated cardiomyocytes could reproduce the tyrosine phosphorylation of P40763 at 5 min . This delayed phosphorylation was almost completely inhibited by anti- P40189 blocking antibody RX435 , but not by TAK044 ( P25101 /B-R antagonist ) , prazosin , or propranolol . AngII induced phosphorylation of P40189 in the late stage , which was temporally in parallel with the delayed phosphorylation of P40763 . AngII augmented P05231 , Q16619 , and P15018 mRNA expression at 30-60 min , but not P26441 expression . AngII increased P05231 protein levels by 3-fold in the conditioned media at 2 h compared with the control . These findings indicated that AngII-induced delayed activation of P40763 is caused by autocrine/paracrine secreted P05231 family cytokines . Inhibition of the signal transducer and activator of transcription-3 ( P40763 ) signaling pathway by 4-oxo-1-phenyl-1,4-dihydroquinoline-3-carboxylic acid esters . The JAK- P40763 pathway regulates genes that are important in cell proliferation and thus is a promising target for cancer therapy . A high-throughput screening ( HTS ) campaign using an Apo-ONE Homogenous Caspase 3/7 assay in U266 cells identified 4-oxo-1-phenyl-1,4-dihydroquinoline-3-carboxylic acid ethyl ester 4 as a potential P40763 pathway inhibitor . Optimization of this HTS hit led to the identification of the 7-cyano analogue 8 , which inhibited P40763 -Y705 phosphorylation with an EC 50 of 170 nM . Compound 8 also inhibited cytokine induced JAK activation but did not inhibit P11274 - P00519 activated P42229 phosphorylation in K562 cells . Chronic delivery of a thrombospondin-1 mimetic decreases skeletal muscle capillarity in mice . Angiogenesis is an essential process for normal skeletal muscle function . There is a growing body of evidence suggesting that thrombospondin-1 ( P07996 -1 ) , a potent antiangiogenic protein in tumorigenesis , is an important regulator of both physiological and pathological skeletal muscle angiogenesis . We tested the hypothesis that chronic exposure to a P07996 -1 mimetic ( DB05434 ) , which targets the P16671 P07996 -1 receptor , would decrease skeletal muscle capillarity as well as alter the balance between positive and negative angiogenic proteins under basal conditions . Osmotic minipumps with either DB05434 or vehicle ( 5 % dextrose ) were implanted subcutaneously in the subscapular region of C57/BL6 mice for 14 days . When compared to the vehicle treated mice , the DB05434 group had a 20 % decrease in capillarity in the superficial region of the gastrocnemius ( GA ) , 11 % decrease in the plantaris ( Q02083 ) , and a 35 % decrease in the soleus ( SOL ) . DB05434 also decreased muscle protein expression of vascular endothelial growth factor ( P15692 ) in both the GA ( -140 % ) and SOL ( -62 % ) ; however there was no change in P15692 in the Q02083 . Serum P15692 was not altered in DB05434 treated animals . Endogenous P07996 -1 protein expression in all muscles remained unaltered . Tunnel staining revealed no difference in muscle apoptosis between DB05434 and vehicle treated groups . These data provide evidence that the anti-angiogenic effects of P07996 -1 are mediated , at least in part , via the P16671 receptor . It also suggests that under physiologic conditions the P07996 -1/ P16671 axis plays a role in regulating basal skeletal muscle microvessel density . Technique appraisement of comparative proteomics and screening of differentiation-related protein in gastric carcinoma . BACKGROUND/AIMS : Different differentiations of cancer have resulted in its unique biological characteristics . We screen and appraise differentially expressed proteins in different differentiated gastric adenocarcinoma with comparative proteomics technology in order to find regulatory factors of tumor differentiation related and finally reach the purpose of tumor differentiation reversal . METHODOLOGY : With two-dimensional fluorescence difference gel electrophoresis ( 2-D DIGE ) and liquid chromatography in conjunction with tandem mass spectrometry ( LC–MS/MS ) , the differentially expressed proteins from 8 patients with different differentiated gastric adenocarcinoma were identified and some factors identified were verified with application of QPCR and Western blot techniques . RESULTS : Significant differences in 35 protein spots were found and 48 kinds of proteins were identified . Other than structural proteins and non-specific protein , six possible proteins associated with tumor differentiation were determined - the serine protease inhibitor B1 ( serine protease inhibitor , clade B , member 1 , P30740 ) , calcium-phospholipid binding protein III ( annexin A3 ) , transcription factor Nm23-H1 , adenine phosphoribosyl-transferase enzyme P07741 ( DB00173 Phosphoribosyltransferase in APO and AMP ) , glutathione S-transferase P1-1 ( Q86UG4 -π-1 ) , antimicrobial peptides P81605 -lL . The identified P30740 , annexin A3 , Nm23-H1 and P07741 were verified , confirming the expression of these factors was in line with proteomics identification . CONCLUSIONS : Protein expression in different differentiated gastric adenocarcinoma was varied . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . SH2B1β interacts with P40763 and enhances fibroblast growth factor 1-induced gene expression during neuronal differentiation . Neurite outgrowth is an essential process during neuronal differentiation as well as neuroregeneration . Thus , understanding the molecular and cellular control of neurite outgrowth will benefit patients with neurological diseases . We have previously shown that overexpression of the signaling adaptor protein SH2B1β promotes fibroblast growth factor 1 ( P05230 ) -induced neurite outgrowth ( W. F. Lin , C. J. Chen , Y. J. Chang , S. L. Chen , I. M. Chiu , and L. Chen , Cell. Signal. 21:1060-1072 , 2009 ) . SH2B1β also undergoes nucleocytoplasmic shuttling and regulates a subset of neurotrophin-induced genes . Although these findings suggest that SH2B1β regulates gene expression , the nuclear role of SH2B1β was not known . In this study , we show that SH2B1β interacts with the transcription factor , signal transducer , and activator of transcription 3 ( P40763 ) in neuronal PC12 cells , cortical neurons , and COS7 fibroblasts . By affecting the subcellular distribution of P40763 , SH2B1β increased serine phosphorylation and the concomitant transcriptional activity of P40763 . As a result , overexpressing SH2B1β enhanced P05230 -induced expression of P40763 target genes Egr1 and Cdh2 . Chromatin immunoprecipitation assays further reveal that , in response to P05230 , overexpression of SH2B1β promotes the in vivo occupancy of P40763 -Sp1 heterodimers at the promoter of Egr1 and Cdh2 . These findings establish a central role of SH2B1β in orchestrating signaling events to transcriptional activation through interacting and regulating P40763 -containing complexes during neuronal differentiation . Greater functional ETB receptor antagonism with DB00559 than sitaxsentan in healthy men . Endothelin ( ET ) -1 is implicated in the development of hypertension and a role for endothelin receptor antagonists ( ETRAs ) in the management of hypertension is emerging . ETRAs are classified as selective or mixed depending on their degree of ET(A):ET(B) receptor blockade . As yet , there are no comparative studies in humans that measure biochemical and functional ET(B) blockade achieved by currently licensed ETRAs . We therefore investigated the effects of DB00559 , a mixed P25101 , and sitaxsentan , an ET(A) selective P25101 , on plasma ET-1 concentrations and ET(B)-mediated vasodilatation to P14138 . In a randomized , double-blind , 3-way crossover study , 10 healthy subjects received 7 days of placebo , DB00559 250 mg , and sitaxsentan 100 mg daily . Plasma ET-1 concentrations were measured at baseline and 3 hours on day 1 and predose on day 7 . Subjects also underwent forearm blood flow measurements on day 7 of each period with brachial artery infusion of P14138 ( 60 pmol/min for 5 minutes ) . DB00559 , but not placebo or sitaxsentan , significantly increased plasma ET-1 concentrations at day 7 ( +0.70+/-0.20 pg/mL ; P < 0.005 ) . Maximal P14138 -mediated vasodilatation was seen at 2 minutes following placebo ( 30+/-6 % ) and sitaxsentan ( 21+/-11 % ) ; however , this was abolished by DB00559 , with a reduction in forearm blood flow of 8+/-3 % ( P < 0.01 versus placebo and sitaxsentan ) . DB00559 but not sitaxsentan increases circulating plasma ET-1 levels and abolishes acute P14138 -mediated vasodilatation , confirming that the mixed ET(A/B) antagonist DB00559 , but not the selective ET(A) antagonist sitaxsentan , causes functional ET(B) blockade at clinically relevant doses in healthy human subjects . P07550 -mediated effects on sinus rate and atrial and ventricular contractility on isolated , blood-perfused dog heart preparations . Changes in the sinus rate , right atrial contractile force and left ventricular contractile force in response to isoproterenol , epinephrine , dobutamine , salbutamol and procaterol were studied in isolated , blood-perfused right atrial or left ventricular cardiac preparations of the dog . Each substance elicited dose-dependent increases in the three parameters and the ranking of the potency ( ED50 ) for each effect was isoproterenol greater than epinephrine greater than dobutamine greater than or equal to salbutamol greater than or equal to procaterol . The ED50 of procaterol for changing sinus rate was lower than for altering atrial and ventricular contractile force , whereas the ED50 of dobutamine for changing sinus rate was higher . Ranking on the basis of the ratio of increase in sinus rate to increase in atrial tension induced by the agonists gave the following order : procaterol greater than or equal to salbutamol greater than epinephrine greater than or equal to isoproterenol greater than dobutamine . DB01366 -induced increases in sinus rate and atrial contractile force were dose-dependently inhibited by the beta-2 adrenoceptor antagonist , ICI 118,551 , but only attenuated slightly by the beta-1 antagonist , atenolol . On the other hand , the positive chrono- and inotropic effects on the right atrium induced by dobutamine and isoproterenol were blocked completely by atenolol . The epinephrine- or salbutamol-induced positive chrono- and inotropic responses in the right atrium were inhibited moderately by both antagonists , but ICI 118,551 inhibited sinus rate increases more effectively than the atrial tension increases. ( ABSTRACT TRUNCATED AT 250 WORDS ) DB00114 values in cerebrospinal fluid : reference values and diagnosis of Q9NVS9 deficiency in paediatric patients . Our aim was to establish reference values for cerebrospinal fluid ( P04141 ) pyridoxal 5'-phosphate ( PLP ) in a paediatric population for the diagnosis of pyridox(am)ine 5'-phosphate oxidase ( Q9NVS9 ) deficiency . For reference values , P04141 samples from 113 paediatric controls ( age range : 1 day-18 years ) from Barcelona and London were analysed . Cerebrospinal fluid PLP and biogenic amine concentrations were analysed by HPLC with fluorescence and electrochemical detection . DB00114 concentrations in 4 patients with Q9NVS9 deficiency were determined . A negative correlation between P04141 PLP values and age of controls was observed in both populations ( r=-0.503 ; p < 0.0001 and r=-0.542 ; p=0.002 ) . Reference values were stratified into 4 ( Barcelona ) and 3 age groups ( London ) . For the newborn period , P04141 PLP reference intervals were 32-78 and 44-89 nmol/L for the Barcelona and London centers , respectively ) . No correlation was observed in the different age groups between PLP values and biogenic amines metabolites . PLP values in neonates with Q9NVS9 deficiency were clearly decreased ( PLP=3.6 , 12.0 , 14.0 and 18.0 nmol/L ) compared with our reference ranges . In conclusion , reference values for P04141 PLP should be stratified according to age . No association was observed between PLP values and biogenic amines metabolites . In our 4 cases with Q9NVS9 deficiency , P04141 PLP values were clearly below the reference values . Selective inhibition of the tumor marker O60218 by antiinflammatory N-phenylanthranilic acids and glycyrrhetic acid . A human aldose reductase-like protein , O60218 in the aldo-keto reductase ( AKR ) superfamily , was recently identified as a tumor marker of several types of cancer . DB02383 , an aldose reductase inhibitor ( Q9Y4X5 ) , is known to be the most potent inhibitor of the enzyme . In this study , we compared the inhibitory effects of other ARIs including flavonoids on O60218 and aldose reductase to evaluate their specificity . However , ARIs showed lower inhibitory potency for O60218 than for aldose reductase . In the search for potent and selective inhibitors of O60218 from other drugs used clinically , we found that non-steroidal antiinflammatory N-phenylanthranilic acids , diclofenac and glycyrrhetic acid competitively inhibited O60218 , showing K(i) values of 0.35-2.9 microM and high selectivity to this enzyme ( 43-57 fold versus aldose reductase ) . Molecular docking studies of mefenamic acid and glycyrrhetic acid in the O60218 -nicotinamide adenine dinucleotide phosphate ( NADP(+) ) complex and site-directed mutagenesis of the putative binding residues suggest that the side chain of Val301 and a hydrogen-bonding network among residues Val301 , Gln114 and Ser304 are important for determining the inhibitory potency and selectivity of the non-steroidal antiinflammatory drugs . Thus , the potent and selective inhibition may be related to the cancer chemopreventive roles of the drugs , and their structural features may facilitate the design of new anti-cancer agents targeting O60218 .

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interacts_with DB06589?

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DB06589 inhibits the activation of P09619 β-expressing astrocytes in the brain metastatic microenvironment of breast cancer cells . Brain metastases occur in more than one-third of metastatic breast cancer patients whose tumors overexpress P04626 or are triple negative . Brain colonization of cancer cells occurs in a unique environment , containing microglia , oligodendrocytes , astrocytes , and neurons . Although a neuroinflammatory response has been documented in brain metastasis , its contribution to cancer progression and therapy remains poorly understood . Using an experimental brain metastasis model , we characterized the brain metastatic microenvironment of brain tropic , P04626 -transfected MDA-MB-231 human breast carcinoma cells ( 231-BR- P04626 ) . A previously unidentified subpopulation of metastasis-associated astrocytes expressing phosphorylated platelet-derived growth factor receptor β ( at tyrosine 751 ; p751- P09619 β ) was identified around perivascular brain micrometastases . p751- P09619 β(+) astrocytes were also identified in human brain metastases from eight craniotomy specimens and in primary cultures of astrocyte-enriched glial cells . Previously , we reported that pazopanib , a multispecific tyrosine kinase inhibitor , prevented the outgrowth of 231-BR- P04626 large brain metastases by 73 % . Here , we evaluated the effect of pazopanib on the brain neuroinflammatory microenvironment . DB06589 treatment resulted in 70 % ( P = 0.023 ) decrease of the p751- P09619 β(+) astrocyte population , at the lowest dose of 30 mg/kg , twice daily . Collectively , the data identify a subpopulation of activated astrocytes in the subclinical perivascular stage of brain metastases and show that they are inhibitable by pazopanib , suggesting its potential to prevent the development of brain micrometastases in breast cancer patients . Gene expression profiling of human kidneys undergoing laparoscopic donor nephrectomy . BACKGROUND AND OBJECTIVES : The objective was to compare gene expression profiles of 6 kidneys from open donor nephrectomy with 6 kidneys removed after laparoscopic donor nephrectomy and several hours of carbon dioxide pneumoperitoneum with DNA microarrays and identify small-molecule drugs . METHODS : The gene expression profile GSE3297 was downloaded from the Gene Expression Omnibus database , and the differentially expressed genes were identified by a bioinformatics approach . First , Osprey software was used to construct a differentially expressed gene associated network . Then , DAVID ( Database for Annotation , Visualization , and Integrated Discovery ) and FuncAssociate were used to perform functional analyses . Finally , the Connectivity Map was used to screen for small-molecule drugs . RESULTS : A total of 285 differentially expressed genes were identified , including 148 down-regulated genes and 137 up-regulated genes . In addition , the differentially expressed genes in the most significant Gene Ontology term were P55212 , P01116 , O15524 , P03372 , P01222 , P02452 , and P50281 . Furthermore , several differentially expressed genes , including P42224 , P42226 , Q07890 , and O15524 , participated in the most remarkable Janus kinase-signal transducer and activator of transcription signaling pathway . Finally , luteolin -- with the highest score ( 0.887 ) -- was identified as the small-molecule drug . CONCLUSIONS : Our data show an altered renal transcriptome induced by several hours of carbon dioxide pneumoperitoneum and laparoscopic surgery characterized by up-regulation of genes associated with acute inflammation , apoptosis , and immune injury , which could potentially result in renal injury and an enhanced immune response in the recipient after transplant . Combination of cetuximab with chemoradiation , trastuzumab or MAPK inhibitors : mechanisms of sensitisation of cervical cancer cells . BACKGROUND : Cervical cancer ( CC ) annually kills 288,000 women worldwide . Unfortunately , responses to chemoradiation are partial and are of short duration . As anti- P00533 monoclonal antibodies sensitise tumours , we investigated cetuximab 's toxicity plus chemoradiation on CC cells , which express different P00533 levels . METHODS : P00533 , P04626 , AKT and MAPK expression and phosphorylation status were determined by western blotting . Cytotoxicity was assessed by MTT or clonogenic assays ( CA ) in cell lines treated with cetuximab alone or in combinations . RESULTS : Cetuximab with cisplatin and radiation achieved maximum cytotoxic effects for A431 , Caski and C33A cells ( high , intermediate and low P00533 expression , respectively ) in CA . Cetuximab efficiently decreased MAPK and AKT phosphorylation in A431 cells but slightly less in Caski and C33A cells . To check whether further P00533 , P04626 or MAPK inhibition would improve cetuximab 's cytotoxicity , we combined it with an P00533 tyrosine kinase inhibitor ( TKI ) , trastuzumab or a Q02750 /2 inhibitor ( PD98059 ) . In Caski , but not in C33A cells , cetuximab cooperated with the TKI , reducing cell survival and AKT and MAPK phosphorylation . However , cetuximab with trastuzumab or PD98059 reduced survival and MAPK phosphorylation of both cell lines . CONCLUSION : Our data suggest that cetuximab combined with chemoradiation , trastuzumab or MAPK inhibitors has useful applications for CC treatment , independently of P00533 expression . Autophagy suppression promotes apoptotic cell death in response to inhibition of the PI3K- P42345 pathway in pancreatic adenocarcinoma . Targeting of pathways downstream of DB01367 represents a promising therapeutic strategy for pancreatic cancer , the fourth leading cause of cancer-related death in the USA , since activation of the Raf-MEK- P29323 and PI3K-AKT pathways is found frequently in this disease and is associated with poor prognosis . Taking advantage of a panel of human PDAC cell lines and specific inhibitors of PI3K and/or P42345 , we systematically address the question whether dual-targeted inhibition of the PI3K and P42345 pathways offers advantages over single-targeted inhibition of PI3K in PDAC . We observe greater overall susceptibility of cell lines to dual inhibition compared to targeting PI3K alone . However , we find that dual inhibition of PI3K and P42345 induces autophagy to a greater extent than inhibition of each target alone . In agreement with this , we show that combined administration of PI3K/ P42345 and autophagy inhibitors results in increased anti-tumor activity in vitro and in vivo in models of pancreatic adenocarcinoma . DB05241 , a PI3K/ P42345 inhibitor used in our in vivo studies , is currently undergoing clinical evaluation in a variety of cancer types , while the autophagy inhibitor chloroquine is a widely used anti-malaria compound . Thus , our studies provide rationale for clinical development of combinations of these compounds for the treatment of pancreatic adenocarcinoma . Modulation of a Mr 175,000 c-neu receptor isoform in Q9UBA6 / P00374 cells by serum starvation . The neu proto-oncogene product has been found to exist in two interconvertible forms in Q9UBA6 / P00374 mouse fibroblasts . The 185-kilodalton form ( p185 ) present in growing cells is replaced by a 175-kilodalton form ( p175 ) under conditions of serum starvation . This low molecular weight form accounts almost exclusively for the phosphotyrosine content of the receptor and is associated with increased tyrosine kinase activity . Addition of serum , platelet-derived growth factor or tumor promoter induces conversion of p175 to p185 within minutes , and this increase in molecular weight is associated with phosphorylation of serine and threonine ; removal of serum growth factors is followed by replacement of p185 with p175 over several hours . Unlike Q9UBA6 / P00374 cells , the human breast cancer cell line SK-Br-3 expresses a high molecular weight neu/ P04626 receptor with unchanged phosphotyrosine content in both serum-starved and serum-stimulated cultures . These findings indicate that activation of the neu proto-oncogene product in Q9UBA6 / P00374 cells may be regulated in part by protein kinase C-mediated receptor transmodulation rather than by ligand availability alone . Kinase inhibitors : a new class of antirheumatic drugs . The outlook for patients with rheumatoid arthritis has improved significantly over the last three decades with the use of disease-modifying antirheumatic drugs . However , despite the use of methotrexate , cytokine inhibitors , and molecules targeting T and B cells , a percentage of patients do not respond or lose their response over time . The autoimmune process in rheumatoid arthritis depends on activation of immune cells , which utilize intracellular kinases to respond to external stimuli such as cytokines , immune complexes , and antigens . In the past decade , small molecules targeting several kinases , such as p38 MAPK , Syk , and JAK have been developed . Several p38 MAPK inhibitors proved ineffective in treating rheumatoid arthritis . The Syk inhibitor , fostamatinib , proved superior to placebo in Phase II trials and is currently under Phase III investigation . DB08895 , a P23458 /3 inhibitor , was shown to be efficacious in two Phase III trials , while VX-509 , a P52333 inhibitor , showed promising results in a Phase II trial . Fostamatinib and tofacitinib were associated with increased rates of infection , elevation of liver enzymes , and neutropenia . Moreover , fostamatinib caused elevations of blood pressure and diarrhea , while tofacitinib was associated with an increase in creatinine and elevation of lipid levels . Comprehensive molecular characterization of urothelial bladder carcinoma . Urothelial carcinoma of the bladder is a common malignancy that causes approximately 150,000 deaths per year worldwide . So far , no molecularly targeted agents have been approved for treatment of the disease . As part of The Cancer Genome Atlas project , we report here an integrated analysis of 131 urothelial carcinomas to provide a comprehensive landscape of molecular alterations . There were statistically significant recurrent mutations in 32 genes , including multiple genes involved in cell-cycle regulation , chromatin regulation , and kinase signalling pathways , as well as 9 genes not previously reported as significantly mutated in any cancer . RNA sequencing revealed four expression subtypes , two of which ( papillary-like and basal/squamous-like ) were also evident in microRNA sequencing and protein data . Whole-genome and RNA sequencing identified recurrent in-frame activating P22607 - Q9Y6A5 fusions and expression or integration of several viruses ( including HPV16 ) that are associated with gene inactivation . Our analyses identified potential therapeutic targets in 69 % of the tumours , including 42 % with targets in the phosphatidylinositol-3-OH kinase/AKT/ P42345 pathway and 45 % with targets ( including P04626 ) in the RTK/MAPK pathway . Chromatin regulatory genes were more frequently mutated in urothelial carcinoma than in any other common cancer studied so far , indicating the future possibility of targeted therapy for chromatin abnormalities . Changes of thyroid hormone levels and related gene expression in zebrafish on early life stage exposure to triadimefon . In this study , zebrafish was exposed to triadimefon . Thyroid hormones levels and the expression of related genes in the hypothalamic-pituitary-thyroid ( Q9HD23 ) axis , including thyroid-stimulating hormone ( P01222 ) , deiodinases ( dio1 and dio2 ) and the thyroid hormone receptor ( thraa and thrb ) were evaluated . After triadimefon exposure , increased DB00451 can be explained by increased thyroid-stimulating hormone ( P01222 ) . The conversion of DB00451 to DB00279 ( deiodinase type I-dio1 ) was decreased , which reduced the DB00279 level . P10828 ( thrb ) mRNA levels were significantly down-regulated , possibly as a response to the decreased DB00279 levels . The overall results indicated that triadimefon exposure could alter gene expression in the Q9HD23 axis and that mechanisms of disruption of thyroid status by triadimefon could occur at several steps in the synthesis , regulation , and action of thyroid hormones . DB08888 for vitreoretinal diseases . P02751 and laminin are clinically relevant plasmin receptors in the eye . Located at the vitreoretinal interface , they are cleaved by ocriplasmin ( DB05028 , ThromboGenics , Iselin , NJ ) , a novel ophthalmic medication . A series of clinical trials to study ocriplasmin for the treatment of vitreoretinal diseases such as vitreomacular traction , macular hole , and exudative age-related macular degeneration are underway . The results are promising and may impact patient care . Expression and mutational status of treatment-relevant targets and key oncogenes in 123 malignant salivary gland tumours . BACKGROUND : Malignant tumours of the salivary glands ( MSGT ) are rare and pleomorphic entities . Patients with advanced disease may benefit from targeted therapy ; however , specific targets for optimising and personalising treatments are yet to be identified . DESIGN : Immunohistochemistry for C- P10721 , P00533 , P04626 , P15941 , phospho- P42345 , androgen/estrogens/progesterone receptors and Ki67 was carried out and evaluated in terms of progression-free and overall survival . High throughput molecular screening of key oncogenes was done in 107 patients using routine diagnostic methods and Sequenom technology . RESULTS : Several therapy leads were identified , including high levels of P04626 and androgen receptors in salivary duct carcinomas , C- P10721 in myoepithelial carcinomas and P00533 in mucoepidermoid carcinomas . Recurrent mutations involving downstream elements of the P00533 pathway were found in P01112 , notably in tumours with a myoepithelial component , and in other key oncogenes ( P01116 / P01111 /PI3KCA/ P15056 /MAP2K ) . On the other hand , < 1 % of samples had P00533 or P04626 mutations . CONCLUSION : Several tumour subtypes overexpressed targets of directed therapies suggesting potential therapy leads . Genotyping results suggest activation downstream of P00533 in 18 of the 107 samples that could be associated with low efficacy of P00533 inhibitors . Other molecules , such as PI3K/MEK or P42345 inhibitors , may have anti-tumour activity in this subgroup . The high mutation rate in P01112 highlights a novel key oncogenic event in MSGT . Neuromuscular therapeutics by RNA-targeted suppression of P22303 gene expression . RNA-targeted therapeutics offers inherent advantages over small molecule drugs wherever one out of several splice variant enzymes should be inhibited . Here , we report the use of Monarsen , a 20-mer acetylcholinesterase-targeted antisense agent with three 3'-2'o-methyl-protected nucleotides , for selectively attenuating the stress-induced accumulation of the normally rare , soluble " readthrough " acetylcholinesterase variant P22303 -R . DB03128 hydrolysis by P22303 -R may cause muscle fatigue and moreover , limit the cholinergic anti-inflammatory blockade , yielding inflammation-associated pathology . Specific P22303 -R targeting by Monarsen was achieved in cultured cells , experimental animals , and patient volunteers . In rats with experimental autoimmune myasthenia gravis , oral delivery of Monarsen improved muscle action potential in a lower dose regimen ( nanomolar versus micromolar ) , rapid and prolonged manner ( up to 72 h versus 2-4 h ) as compared with the currently used small molecule anticholinesterases . In central nervous system neurons of both rats and cynomolgus monkeys , systematic Monarsen treatment further suppressed the levels of the proinflammatory cytokines interleukin-1 ( IL-1 ) and P05231 . Toxicology testing and ongoing clinical trials support the notion that Monarsen treatment would offer considerable advantages over conventional cholinesterase inhibitors with respect to dosing , specificity , side effects profile , and duration of efficacy , while raising some open questions regarding its detailed mechanism of action . Neuregulin-1 activates the JAK- P35610 pathway and regulates lung epithelial cell proliferation . Neuregulin-1 ( Q99988 ) is part of a family of proteins whose members are structurally related to epidermal growth factor . Q99988 induces cell proliferation through a high-affinity receptor complex composed of a heterodimer of human epidermal growth factor-like receptor ( HER ) 2 and 3 . In this study , we show that Q99988 activates the Janus kinases ( JAK ) and signal transducer and activator of transcription proteins ( P35610 ) . Q99988 induced a rapid and transient increase in tyrosine phosphorylation of P29597 and P52333 , but not P23458 or O60674 , and induced P40763 and P42229 tyrosine phosphorylation . Upon phosphorylation , P40763 translocated to the nucleus within 1 h . Activation of the JAK- P35610 pathway was dependent on P04626 / P21860 heterodimerization and was necessary for Q99988 -induced proliferation . Inhibition of P04626 's ability to dimerize using the P04626 -specific antibody 2C4 completely blocked Q99988 -induced P52333 , P29597 , P40763 , and P42229 tyrosine phosphorylation . Blocking the JAK- P35610 pathway with a specific JAK- P35610 pathway inhibitor , AG490 , inhibited Q99988 -induced JAK and P35610 phosphorylation and cell proliferation . These data suggest that Q99988 activates the JAK- P35610 signal transduction pathway through its high-affinity receptor , the P04626 / P21860 heterodimer . This pathway plays an important role in Q99988 -stimulated proliferation of pulmonary epithelial cells . Absolute bioavailability and effect of formulation change , food , or elevated pH with rabeprazole on cobimetinib absorption in healthy subjects . DB05239 is a potent and highly selective inhibitor of Q02750 /2 . Since cobimetinib exhibited absorption variability in cancer patients , a series of single-dose studies in healthy subjects were conducted to determine absolute bioavailability and elucidate potential effects of formulation , food , and elevated gastric pH on cobimetinib bioavailability . Three crossover trials were performed with a 20 mg cobimetinib oral dose : absolute bioavailability using a 2 mg intravenous infusion ( n = 13 ) , relative bioavailability of tablets versus capsules and food effect ( n = 20 ) , and drug interaction with a proton pump inhibitor ( 20 mg of rabeprazole daily for 5 days prior to cobimetinib administration ; n = 20 ) . Absolute bioavailability of cobimetinib was 46.2 % ( 24.2 , CV % ) , likely due to metabolism rather than incomplete absorption . The mean systemic clearance of cobimetinib was low ( 11.7 L/h [ 28.2 , CV % ] ) . Administration of cobimetinib tablets with a high-fat meal delayed drug absorption ( prolonged tmax ) but had no statistically significant effect on cobimetinib exposure ( Cmax and AUC0-∞ ) . Tablet and capsule formulations of cobimetinib showed comparable exposures . DB05239 exhibited delayed absorption ( tmax ) in the presence of rabeprazole , with no statistically significant effects on drug exposure ( Cmax and AUC0-∞ ) in the fasted state . In conclusion , cobimetinib oral absorption was not affected by change in formulation , food , or elevated gastric pH . cDNA microarray analysis of genes associated with P04626 ( P04626 /neu ) overexpression in human mammary luminal epithelial cells . To investigate changes in gene expression associated with P04626 , expression profiling of immortalized human mammary luminal epithelial cells and variants expressing a moderate and high level of P04626 has been carried out using cDNA microarrays corresponding to approximately 6000 unique genes/ESTs . A total of 61 significantly up- or downregulated ( 2.0-fold ) genes were identified and further validated by RT-PCR analysis as well as microarray comparisons with a spontaneously P04626 - overexpressing breast cancer cell line and P04626 -positive primary breast tumors . The expression and clinical relevance of proteins predicted to be associated with P04626 overexpression in breast cancers were analysed together with their clinical relevance by antibody screening using a tissue array . Differentially regulated genes include those involved in cell-matrix interactions including proline 4-hydroxylase ( O15460 ) , galectin 1 ( P09382 ) and galectin 3 ( P17931 ) , fibronectin 1 ( P02751 ) and p-cadherin ( CDH3 ) , and cell proliferation ( P50238 , P17936 ) and transformation ( P25815 , P26447 ) . A number of genes associated with MYC signalling were also differentially expressed , including Q92597 , Q15853 and the epithelial membrane proteins 1 and 3 ( P54849 , P54852 ) . These data represent profiles of the transcriptional changes associated with P04626 -related pathways in the breast , and identify novel and potentially useful targets for prognosis and therapy . Phosphodiesterases Regulate BAY 41-2272-Induced P50552 Phosphorylation in Vascular Smooth Muscle Cells . BAY 41-2272 ( BAY ) , a stimulator of soluble guanylyl cyclase , increases cyclic nucleotides and inhibits proliferation of vascular smooth muscle cells ( VSMCs ) . In this study , we elucidated mechanisms of action of BAY in its regulation of vasodilator-stimulated phosphoprotein ( P50552 ) with an emphasis on VSMC phosphodiesterases ( PDEs ) . BAY alone increased phosphorylation of P50552 (Ser239) and P50552 (Ser157) , respective indicators of PKG and PKA signaling . DB07954 , a non-selective inhibitor of PDEs , had no effect on BAY-induced phosphorylation at P50552 (Ser239) but inhibited phosphorylation at P50552 (Ser157) . Selective inhibitors of PDE3 or DB05876 attenuated BAY-mediated increases at P50552 (Ser239) and P50552 (Ser157) , whereas O76074 inhibition potentiated BAY-mediated increases only at P50552 (Ser157) . In comparison , 8Br-cGMP increased phosphorylation at P50552 (Ser239) and P50552 (Ser157) which were not affected by selective PDE inhibitors . In the presence of 8Br- DB02527 , inhibition of either DB05876 or O76074 decreased P50552 (Ser239) phosphorylation and inhibition of PDE3 increased phosphorylation at P50552 (Ser239) , while inhibition of PDE3 or DB05876 increased and O76074 inhibition had no effect on P50552 (Ser157) phosphorylation . These findings demonstrate that BAY operates via DB02527 and cGMP along with regulation by PDEs to phosphorylate P50552 in VSMCs and that the mechanism of action of BAY in VSMCs is different from that of direct cyclic nucleotide analogs with respect to P50552 phosphorylation and the involvement of PDEs . Given a role for P50552 as a critical cytoskeletal protein , these findings provide evidence for BAY as a regulator of VSMC growth and a potential therapeutic agent against vasculoproliferative disorders . Changes in enzyme activity and expression of P00374 of Toxoplasma gondii by antifolates . The responses to antifolates of Toxoplasma gondii were investigated by measuring the dihydrofolate reductase ( P00374 ) activity , quantity of P00374 mRNA , and single-strand conformational polymorphism ( SSCP ) pattern . DB00205 ( Q9BRP8 ) and methotrexate ( MTX ) were tested as antifolates . When T. gondii was treated with Q9BRP8 , the viability was decreased by the increasing concentration of Q9BRP8 , P00374 activity tended to increase as the passage proceeded , and the quantity of mRNA expressed was also increased according to passages . The viability of T. gondii was decreased by the increasing concentration of MTX , but it was maintained over 40 % up to 100 microM MTX . P00374 activity was 77.4 % in the 1st passage ( 1 microM ) . 82.2 % in the 4th passage ( 10 microM ) , and 141.3 % in the 7th passage ( 100 microM ) . But no changes were detected in SSCP pattern of T. gondii exposed to Q9BRP8 and MTX , both . These results suggested that the response of T. gondii to Q9BRP8 was regulated by transcriptional level and that , in MTX , the viability of T. gondii was derived from increasing P00374 activity . Mechanism of oral absorbent DB05269 in lipid abnormalities in experimental uremic rats . BACKGROUND : We have reported that oral sorbent DB05269 ( Q9NRA2 ) is effective in delaying the induction of dialysis in patients with chronic renal failure ( CRF ) because of its effect on lipid metabolism . To clarify the precise mechanism of Q9NRA2 in lipid abnormalities in CRF , we examined the effect of Q9NRA2 on plasma lipid profile , total bile acids ( TBA ) , and lipoprotein lipase ( P06858 ) activity in experimental uremic rats . METHODS : Uremic rats were prepared using male Wistar rats by ligating 5/6 of the renal artery . Uremic rats were randomly divided into two groups as follows : a control group in which rats were maintained on the standard diet and an Q9NRA2 group in which rats were maintained on a diet containing 5 g of Q9NRA2 per 100 g of standard diet for 10 weeks . Plasma P06858 activity was measured as free fatty acid ( FFA ) generation after intravenous administration of heparin . RESULTS : Plasma creatinine at 1.5 +/- 0.1 mg/dl was lower in the Q9NRA2 group than the 1.9 +/- 0.5 mg/ml level in the control group . Q9NRA2 significantly decreased plasma total cholesterol from 192 +/- 29 to 142 +/- 25 mg/dl , triglycerides from 198 +/- 71 to 99 +/- 38 mg/dl , and TBA from 19.6 +/- 2.6 mumol/liter to 8.8 +/- 3.5 mumol/ml . Plasma P06858 activity at 0.22 +/- 0.01 mumol FFA/min/hr was significantly higher in the Q9NRA2 group than 0.15 +/- 0.03 mumol FFA/min/hr in the control group . CONCLUSIONS : These results suggest that Q9NRA2 may improve plasma lipid abnormalities by binding to bile acids in the intestinal lumen and preventing their reabsorption and inhibiting the reduction of P06858 activity in experimental uremic rats . The cooperation between hMena overexpression and P04626 signalling in breast cancer . hMena and the epithelial specific isoform hMena(11a) are actin cytoskeleton regulatory proteins belonging to the Ena/ P50552 family . P01133 treatment of breast cancer cell lines upregulates hMena/hMena(11a) expression and phosphorylates hMena(11a) , suggesting cross-talk between the ErbB receptor family and hMena/hMena(11a) in breast cancer . The aim of this study was to determine whether the hMena/hMena(11a) overexpression cooperates with HER-2 signalling , thereby affecting the P04626 mitogenic activity in breast cancer . In a cohort of breast cancer tissue samples a significant correlation among hMena , P04626 overexpression , the proliferation index ( high Ki67 ) , and phosphorylated MAPK and AKT was found and among the molecular subtypes the highest frequency of hMena overexpressing tumors was found in the P04626 subtype . From a clinical viewpoint , concomitant overexpression of P04626 and hMena identifies a subgroup of breast cancer patients showing the worst prognosis , indicating that hMena overexpression adds prognostic information to P04626 overexpressing tumors . To identify a functional link between P04626 and hMena , we show here that P04626 transfection in MCF7 cells increased hMena/hMena(11a) expression and hMena(11a) phosphorylation . On the other hand , hMena/hMena(11a) knock-down reduced P21860 , AKT and Q8TCB0 /42 MAPK phosphorylation and inhibited the P01133 and Q02297 -dependent P04626 phosphorylation and cell proliferation . Of functional significance , hMena/hMena(11a) knock-down reduced the mitogenic activity of P01133 and Q02297 . Collectively these data provide new insights into the relevance of hMena and hMena(11a) as downstream effectors of the ErbB receptor family which may represent a novel prognostic indicator in breast cancer progression , helping to stratify patients . Hypoxic/normoxic preconditioning increases endothelial differentiation potential of human bone marrow CD133+ cells . CD133+ cells are hemangioblasts that have capacity to generate into both hematopoietic and endothelial cells ( ECs ) . Hypoxia/normoxia has shown to be the regulator of the balance between stemness and differentiation . In this study we performed Agilent 's whole human genome oligo microarray analysis and examined the differentiation potential of the bone-marrow-derived CD133+ cells after hypoxic/normoxic preconditioning of CD133+ cells . Results showed that there was no significant increase in erythroid colony forming unit ( CFU-E ) and CFU-granulocyte , erythrocyte , monocyte , and megakaryocyte formation with cells treated under hypoxia/normoxia . However , a significant increment of EC forming unit at 24 h ( 143.2 +/- 8.0 % ) compared to 0 h ( 100 +/- 11.4 % ) was observed in CFU-EC analysis . Reverse transcription-polymerase chain reaction and immunostaining analysis showed that the differentiated cells diminished hematopoietic stem cell surface markers and acquired the gene markers and functional phenotype of ECs . The transcriptome profile revealed a cluster of 232 downregulated and 498 upregulated genes in cells treated for 24 h under hypoxia . The upregulated genes include angiogenic genes , angiogenic growth factor genes , angiogenic cytokine and chemokine genes , as well as angiogenic-positive regulatory genes , including Q14512 , PDGFB , Q16663 , P48061 , P80162 , P05231 , P21246 , O14944 , P04626 , O95136 , P11487 , Q92913 , Q99988 , P05412 , L1CAM , Q02297 , P08138 , and PDGFB . On the other hand , angiogenesis inhibitors and related genes , including P29459 , P98177 , Q9NY15 , and P16035 , are downregulated . Taken together , hypoxic/normoxic preconditioning may lead to the differentiation of CD133+ cells toward endothelial lineage , which may improve the current clinical trial studies . Array-comparative genomic hybridization to detect genomewide changes in microdissected primary and metastatic oral squamous cell carcinomas . Oral squamous cell carcinoma ( OSCC ) is a common worldwide malignancy . However , it is unclear what , if any , genomic alterations occur as the disease progresses to invasive and metastatic OSCC . This study used genomewide array-CGH in microdissected specimens to map genetic alterations found in primary OSCC and neck lymph node metastases . We used array-based comparative genomic hybridization ( array-CGH ) to screen genomewide alterations in eight pairs of microdissected tissue samples from primary and metastatic OSCC . In addition , 25 primary and metastatic OSCC tissue pairs were examined with immunohistochemistry for protein expression of the most frequently altered genes . The highest frequencies of gains were detected in P12524 , Q04864 , TERC , P42336 , P10242 , P08183 , P01112 , GARP , P30279 , P07332 , P04626 , P01127 , and Q05066 . The highest frequencies of losses were detected in p44S10 , O15164 , P06858 , Q13126 , P35226 , P11161 , and Q13163 . Genomic alterations in TGFbeta2 , cellular retinoid-binding protein 1 gene ( P09455 ) , P42336 , P28222 , P01112 , P21860 , and O14965 differed significantly between primary OSCC and their metastatic counterparts . Genomic alterations in Q05513 , P00519 , and P08620 were significantly different in patients who died compared with those who survived . Immunohistochemistry confirmed high P42336 immunoreactivity in primary and metastatic OSCC . Higher P08620 immunoreactivity in primary OSCC is associated with a worse prognosis . Loss of P09455 immunoreactivity is evident in primary and metastatic OSCC . Our study suggests that precise genomic profiling can be useful in determining gene number changes in OSCC . As our understanding of these changes grow , this profiling may become a practical tool for clinical evaluation .

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Arsenic decreases antinociceptive activity of paracetamol : possible involvement of serotonergic and endocannabinoid receptors . We assessed whether repeated arsenic exposure can decrease paracetamol-mediated antinociception by modulating serotonergic and endocannabinoid pathways . Rats were preexposed to elemental arsenic ( 4ppm ) as sodium arsenite through drinking water for 28 days . Next day paracetamol 's ( 400mg/kg , oral ) antinociceptive activity was assessed through formalin-induced nociception . Serotonin content and gene expression of P08908 , 5- Q13049 and P21554 receptors were evaluated in brainstem and frontal cortex . Arsenic decreased paracetamol-mediated analgesia . DB00316 , but not arsenic , increased serotonin content in these regions . Arsenic attenuated paracetamol-mediated increase in serotonin level . DB00316 did not alter P08908 expression , but caused down-regulation of 5- Q13049 and up-regulation of P21554 receptors . Arsenic down-regulated these receptors . However , paracetamol-mediated down-regulation of 5- Q13049 was more pronounced . Arsenic did not modify paracetamol 's effect on P08908 expression , but reduced paracetamol-mediated down-regulation of 5- Q13049 and reversed up-regulation of P21554 receptors . Results suggest arsenic reduced paracetamol-induced analgesia possibly by interfering with pronociceptive 5- Q13049 and antinociceptive P21554 receptors . Clinical and genetic factors associated with nausea and vomiting in cancer patients receiving opioids . BACKGROUND : This study investigates whether demographical , disease-related and genetic factors contribute to inter-individual differences in nausea and vomiting among patients receiving opioids for cancer pain . METHODS : Cancer patients receiving opioids were included from 17 centres in 11 European countries . Intensities of nausea and vomiting were reported by 1579 patients on four-point categorical scales . In stratified regression models including demographical and disease-related factors as covariates , 96 single nucleotide polymorphisms ( SNPs ) in 16 candidate genes related to opioid- or nausea/vomiting signalling pathways ( P08183 , P35372 , P41145 , P32121 , P42226 , P21964 , P20309 , P08912 , P35367 , P14416 , P35462 , P25103 , P46098 , O95264 , Q8WXA8 , P21554 ) were analysed for association with nausea and vomiting . FINDINGS : Age , body mass index , Karnofsky Performance Status , gender , use of antiemetics , type of opioid , type of cancer and eight SNPs were associated with the inter-individual differences in nausea and vomiting among cancer patients treated with opioids ( p < 0.01 ) . The SNPs were rs1176744 , rs3782025 and rs1672717 in O95264 ; rs165722 , rs4680 and rs4633 in P21964 ; rs10802789 and rs685550 in P20309 . Only the SNP rs1672717 in O95264 passed the Benjamini-Hochberg criterion for a 10 % false discovery rate . INTERPRETATION : Clinical characteristics and SNPs within the O95264 , P21964 and P20309 genes may be associated with the variability in nausea and vomiting among cancer patients receiving opioids . This knowledge may help to identify patients at particular risk for nausea and vomiting during treatment with opioids for cancer pain . Met326Ile aminoacid polymorphism in the human p85 alpha gene has no major impact on early insulin signaling in type 2 diabetes . Class I alpha phosphatidylinositol ( PI ) 3-kinase is an important enzyme in the early insulin signaling cascade , and plays a key role in insulin-mediated glucose transport . Despite extensive investigation , the genes responsible for the development of the common forms of type 2 diabetes remain unknown . This study was performed to identify variants in the coding region of p85 alpha , the regulatory subunit of PI 3-kinase . Fibroblasts from skin biopsies from type 2 diabetics and controls were established to address this issue . P85 alpha cDNA was sequenced , and a single point mutation at codon 326 was found . This mutation resulted in a homozygous missense amino acid change DB00134 --> DB00167 in one subject with type 2 diabetes and heterozygous variant in two other diabetic patients and one with severe insulin resistance . Interestingly , those patients revealed an impaired insulin-mediated insulin receptor substrate ( P41252 ) -1 binding to p85 alpha without any alteration in Q9Y4H2 /p85 alpha association . Furthermore , P35568 , Q9Y4H2 , p85 alpha and MAPK protein contents were not significantly changed , and neither were MAPK or Akt phosphorylation . We conclude from our data that this variant may have only minor impact on signaling events ; however , in combination with variants in other genes encoding signaling proteins , this may have a functional impact on early insulin signaling . Inhibition of Niemann-Pick-type C1-like1 by ezetimibe activates autophagy in human hepatocytes and reduces mutant α1-antitrypsin Z deposition . Autophagy can degrade aggregate-prone proteins , but excessive autophagy can have adverse effects . It would be beneficial if autophagy could be enhanced in a cell type-specific manner , but this has been difficult because the basic mechanism of autophagy is common . In the present study we found that inhibition of Niemann-Pick-type C1-like 1 ( Q9UHC9 ) by ezetimibe activates autophagy only in hepatocytes and small intestinal epithelia , but not in other cells . DB00973 induced accumulation of free cholesterol in the late endosome/lysosome and increased partitioning of a Ragulator component , Q6IAA8 , in rafts . The latter change led to down-regulation of mammalian target of rapamycin ( P42345 )C1 activity by decreasing P42345 recruitment to the late endosome/lysosome and activated autophagy . A primary effect of ezetimibe was found to be a decrease of free cholesterol in the plasma membrane , because all the results caused by ezetimibe were suppressed by supplementation of cholesterol as a methyl-β-cyclodextrin complex . By enhancing autophagy in human primary hepatocytes with ezetimibe , insoluble mutant α1-antitrypsin Z was reduced significantly . CONCLUSION : Inhibition of Q9UHC9 by ezetimibe activates autophagy in human hepatocytes by modulating cholesterol homeostasis . DB00973 may be used to ameliorate liver degeneration in α1-antitrypsin deficiency . Characterization of antihistamines using biphasic cutaneous reaction in BALB/c mice . Effects of 11 histamine H1 receptor antagonists on IgE-mediated biphasic cutaneous reaction in mice were examined . The immediate phase reaction ( IPR ) assessed at 1 hour after antigen application was significantly inhibited by all antihistamines examined . The inhibition of IPR by cetirizine and mequitazine were potent , but those by cyproheptadine and diphenhydramine were weak . The later phase reaction ( LPR ) assessed at 24 hours after antigen application was inhibited by chlorpheniramine , oxatomide , ketotifen , mequitazine , emedastine , terfenadine and azelastine . The inhibition of LPR by emedastine was potent , but those by ketotifen and terfenadine were only partial . DB01084 inhibited both IPR and LPR comparably . Present results indicate that H1 receptor activation is involved in the IPR of the biphasic cutaneous reaction , and that the blockade of H1 receptors at IPR does not contribute to the attenuation of following LPR . P35367 antagonists inhibiting the LPR have a property distinct from H1 receptor antagonism , which may have an additional benefit for the treatment of allergic diseases . DB08889 can induce tumor cell death through selective inhibition of the chymotrypsin-like activity of the proteasome . DB08889 is a proteasome inhibitor in clinical development that primarily targets the chymotrypsin-like ( CT-L ) subunits in both the constitutive proteasome ( c20S ) and the immunoproteasome ( i20S ) . To investigate the impact of inhibiting the CT-L activity with carfilzomib , we set out to quantitate the levels of CT-L subunits beta5 from the c20S and P28062 from the i20S in normal and malignant hematopoietic cells . We found that the i20S is a major form of the proteasome expressed in cells of hematopoietic origin , including multiple myeloma ( MM ) CD138+ tumor cells . Although specific inhibition of either P28062 or beta5 alone was insufficient to produce an antitumor response , inhibition of all proteasome subunits was cytotoxic to both hematologic tumor cells and peripheral blood mononuclear cells . However , selective inhibition of both beta5 and P28062 was sufficient to induce an antitumor effect in MM , non-Hodgkin lymphoma , and leukemia cells while minimizing the toxicity toward nontransformed cells . In MM tumor cells , CT-L inhibition alone was sufficient to induce proapoptotic sequelae , including proteasome substrate accumulation , Noxa and caspase 3/7 induction , and phospho-eIF2alpha suppression . These data support a hypothesis that hematologic tumor cells are uniquely sensitive to CT-L inhibition and provide a mechanistic understanding of the clinical safety profile and antitumor activity of proteasome inhibitors . Enhanced killing of cancer cells by poly(ADP-ribose) polymerase inhibitors and topoisomerase I inhibitors reflects poisoning of both enzymes . Poly(ADP-ribose) polymerase-1 ( P09874 ) plays critical roles in the regulation of DNA repair . Accordingly , small molecule inhibitors of PARP are being developed as agents that could modulate the activity of genotoxic chemotherapy , such as topoisomerase I poisons . In this study we evaluated the ability of the PARP inhibitor veliparib to enhance the cytotoxicity of the topoisomerase I poisons topotecan and camptothecin ( CPT ) . DB07232 increased the cell cycle and cytotoxic effects of topotecan in multiple cell line models . Importantly , this sensitization occurred at veliparib concentrations far below those required to substantially inhibit poly(ADP-ribose) polymer synthesis and at least an order of magnitude lower than those involved in selective killing of homologous recombination-deficient cells . Further studies demonstrated that veliparib enhanced the effects of CPT in wild-type mouse embryonic fibroblasts ( MEFs ) but not Parp1(-/-) MEFs , confirming that P09874 is the critical target for this sensitization . Importantly , parental and Parp1(-/-) MEFs had indistinguishable CPT sensitivities , ruling out models in which P09874 catalytic activity plays a role in protecting cells from topoisomerase I poisons . To the contrary , cells were sensitized to CPT in a veliparib-independent manner upon transfection with P09874 E988K , which lacks catalytic activity , or the isolated P09874 DNA binding domain . These results are consistent with a model in which small molecule inhibitors convert P09874 into a protein that potentiates the effects of topoisomerase I poisons by binding to damaged DNA and preventing its normal repair . P04150 antagonism disrupts the reconsolidation of social reward-related memories in rats . Reconsolidation is the process whereby consolidated memories are destabilized upon retrieval and restabilized to persist for later use . Although the neurobiology of the reconsolidation of both appetitive and aversive memories has been intensively investigated , reconsolidation of memories of physiologically relevant social rewards has received little attention . Social play , the most characteristic social behaviour displayed by young mammals , is highly rewarding , illustrated by the fact that it can induce conditioned place preference ( CPP ) . Here , we investigated the role of signalling mechanisms implicated in memory processes , including reconsolidation , namely glucocorticoid , mineralocorticoid , DB01221 glutamatergic and P21554 cannabinoid receptors , in the reconsolidation of social play-induced CPP in rats . Systemic treatment with the glucocorticoid receptor antagonist mifepristone before , but not immediately after , retrieval disrupted the reconsolidation of social play-induced CPP . DB00834 did not affect social play-induced CPP in the absence of memory retrieval . Treatment with the DB01221 receptor antagonist MK-801 modestly affected the reconsolidation of social play-induced CPP . However , the reconsolidation of social play-induced CPP was not affected by treatment with the mineralocorticoid and P21554 cannabinoid receptor antagonists spironolactone and rimonabant , respectively . We conclude that glucocorticoid neurotransmission mediates the reconsolidation of social reward-related memories in rats . These data indicate that the neural mechanisms of the reconsolidation of social reward-related memories only partially overlap with those underlying the reconsolidation of other reward-related memories . Role of antispasmodics in the treatment of irritable bowel syndrome . Irritable bowel syndrome ( IBS ) is a long-lasting , relapsing disorder characterized by abdominal pain/discomfort and altered bowel habits . Intestinal motility impairment and visceral hypersensitivity are the key factors among its multifactorial pathogenesis , both of which require effective treatment . Voltage-gated calcium channels mediate smooth muscle contraction and endocrine secretion and play important roles in neuronal transmission . Antispasmodics are a group of drugs that have been used in the treatment of IBS for decades . DB01616 citrate , a spasmolytic , decreases the sensitivity of smooth muscle contractile proteins to calcium , and it is a selective P08908 receptor antagonist . DB01616 , in combination with simethicone , has been demonstrated to effectively reduce abdominal pain and discomfort in a large placebo-controlled trial . Mebeverine is a musculotropic agent that potently blocks intestinal peristalsis . Non-placebo-controlled trials have shown positive effects of mebeverine in IBS regarding symptom control ; nevertheless , in recent placebo-controlled studies , mebeverine did not exhibit superiority over placebo . Otilonium bromide is poorly absorbed from the GI tract , where it acts locally as an L-type calcium channel blocker , an antimuscarinic and a tachykinin NK2 receptor antagonist . Otilonium has effectively reduced pain and improved defecation alterations in placebo-controlled trials in IBS patients . DB09090 bromide is also an L-type calcium channel blocker that acts locally in the GI tract . DB09090 improves motility disorders and consequently reduces stool problems in IBS patients . Phloroglucinol and trimethylphloroglucinol are non-specific antispasmodics that reduced pain in IBS patients in a placebo-controlled trial . Antispasmodics have excellent safety profiles . T-type calcium channel blockers can abolish visceral hypersensitivity in animal models , which makes them potential candidates for the development of novel therapeutic agents in the treatment of IBS . Antiinflammatory steroid action in human ovarian surface epithelial cells . The human ovarian surface epithelium ( OSE ) is subject to serial injury and repair during ovulation , which is a natural inflammatory event . We asked whether there is a compensatory antiinflammatory component to this process , involving steroid hormones produced locally at the time of ovulation . Quantitative RT-PCR analysis of total RNA from cultured human OSE cell monolayers showed that exposure to proinflammatory IL1alpha ( 500 pg/ml ) increased mRNA levels of cyclooxygenase-2 ( P35354 ) ( P < 0.01 ) at 48 h . The P35354 mRNA response to IL1alpha was associated with an approximate 18-fold ( P < 0.01 ) increase in mRNA levels of 11beta-hydroxysteroid dehydrogenase type 1 ( 11betaHSD1 ) , encoding the steroid dehydrogenase that reversibly reduces cortisone to antiinflammatory cortisol . Addition of cortisol to OSE cell culture medium dose-dependently suppressed the P35354 mRNA response to IL1alpha ( P < 0.01 ) but reciprocally enhanced the 11betaHSD1 mRNA response ( P < 0.05 ) , with both effects strongest at 1 microm cortisol . Presence of glucocorticoid receptor-alpha mRNA and protein was established in OSE cell monolayers and treatment with IL1alpha shown to significantly up-regulate the glucocorticoid receptor-alpha mRNA level ( P < 0.05 ) . P04150 antagonist ( DB00834 , 10 microm ) fully reversed the inhibitory effect of 1 microm cortisol on IL1alpha-stimulated P35354 mRNA expression . Progesterone also suppressed IL1alpha-induced P35354 mRNA expression but had no significant effect on IL1alpha-stimulated 11betaHSD1 expression . These data provide direct evidence for antiinflammatory actions of cortisol and progesterone in human OSE cells . Specific Biomarkers Are Associated with Docetaxeland Gemcitabine-Resistant NSCLC Cell Lines . Five-year survival rate for lung cancer is limited to 10 % to 15 % . Therefore , the identification of novel therapeutic prognostic factors is an urgent requirement . The aim of this study is thus to highlight specific biomarkers in chemoresistant non-small cell lung cancer cell lines . Therefore , we checked-in the control condition as well as after short-term pharmacological treatment with either docetaxel or gemcitabine-the expression of genes such as tumor suppressor genes ( CDKN2A , P53355 , P49789 , P09211 , P16455 , RARβ2 , RASSF1A , and P35625 ) , genes associated with drug resistance ( P38398 , P35354 , P07992 , P17936 , P23921 , and Q13509 ) , and stemness-related genes ( CD133 , Q01860 , and O43623 ) in two cellular models of squamous carcinoma ( CAEP ) and adenocarcinoma ( RAL ) of the lung originally established . Their promoter methylation profile was also evaluated . Drug-related genes were upregulated . DB00515 resistance matched with high levels of P38398 and P07992 in both cell lines ; docetaxel sensitivity of CAEP cells was associated to levels of Q13509 lower than RAL cells . Although CAEP cells were more sensitive to gemcitabine , both cell lines showed high levels of P23921 . Stemness-related genes were downregulated in the control condition but became upregulated in docetaxel-resistant cells , indicating the selection of a population with stemness features . We did not find an unequivocal correspondence between gene expression and respective DNA promoter methylation status , suggesting the involvement of additional mechanisms of gene expression regulation . These results highlight specific biomarkers consistent with the different responses of the two cell lines to standard pharmacological treatments and indicate specific molecular traits for their chemoresistance . Combined preoperative use of celecoxib and gabapentin in the management of postoperative pain . BACKGROUND : In 2005 we reported a study on the efficacy of the preoperative use of the selective P35354 inhibitor celecoxib ( DB00482 ) for reducing both postoperative pain and opioid requirements in patients undergoing bilateral subpectoral breast augmentation . Our findings showed that patients who received 400 mg of celecoxib 30 min before surgery required significantly less postoperative opioid analgesics compared with those given a placebo . DB00996 ( DB00996 ) is an agent commonly used to control neuropathic pain . Here we describe a prospective study assessing the efficacy of preoperative gabapentin in combination with celecoxib for reducing postoperative pain and opioid requirements in elective subpectoral breast augmentation . METHODS : One hundred eighteen patients were given 1200 mg of gabapentin and 400 mg of celecoxib 30-60 min before surgery . From the day of surgery until postoperative day 5 , patients documented any use of analgesics and recorded their degree of pain . Results were then compared with those of our previous study in which only celecoxib was used . RESULTS : The combination of gabapentin and celecoxib was found to be significantly superior ( p < 0.001 ) in reducing postoperative pain and opioid requirements than celecoxib alone in the management of postoperative pain and opioid requirements . CONCLUSION : To decrease postoperative opioid requirements , we recommend 400 mg of celecoxib and 1200 mg of gabapentin taken 30-60 min before surgery by patients undergoing subpectoral breast augmentation or a comparable plastic surgery procedure . Pharmacological characterization of 3-[3-tert-butylsulfanyl-1-[4-(6-methoxy-pyridin-3-yl)-benzyl]-5-(pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid ( DB05225 ) , a novel selective P09917 -activating protein inhibitor that reduces acute and chronic inflammation . Leukotrienes ( LTs ) are proinflammatory lipid mediators synthesized by the conversion of arachidonic acid ( AA ) to P01374 (4) by the enzyme P09917 ( P09917 ) in the presence of P09917 -activating protein ( P20292 ) . 3-[3-tert-Butylsulfanyl-1-[4-(6-methoxy-pyridin-3-yl)-benzyl]-5-(pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid ( DB05225 ) is a novel selective P20292 inhibitor in development for the treatment of respiratory conditions such as asthma . In a rat ex vivo whole-blood calcium ionophore-induced Q06643 (4) assay , DB05225 ( administered orally at 1 mg/kg ) displayed > 50 % inhibition for up to 6 h with a calculated EC(50) of approximately 60 nM . When rat lung was challenged in vivo with calcium ionophore , DB05225 inhibited Q06643 (4) and cysteinyl leukotriene ( CysLT ) production with ED(50) values of 0.8 and 1 mg/kg , respectively . In this model , the EC(50) derived from plasma DB05225 was approximately 330 nM for inhibition of both Q06643 (4) and CysLT . In an acute inflammation setting , DB05225 displayed dose-dependent inhibition of Q06643 (4) , CysLT , and plasma protein extravasation induced by peritoneal zymosan injection . In a model of chronic lung inflammation using ovalbumin-primed and challenged BALB/c mice , DB05225 reduced the concentrations of eosinophil peroxidase , CysLTs , and interleukin-5 in the bronchoalveolar lavage fluid . Finally , DB05225 increased survival time in mice exposed to a lethal intravenous injection of platelet-activating factor . In summary , DB05225 is a novel , potent and selective P20292 inhibitor that has excellent pharmacodynamic properties in vivo and is effective in animal models of acute and chronic inflammation and in a model of lethal shock . Ligand-specific dynamics of the progesterone receptor in living cells and during chromatin remodeling in vitro . P06401 ( PR ) , a member of the nuclear receptor superfamily , is a key regulator of several processes in reproductive function . We have studied the dynamics of the interaction of PR with a natural target promoter in living cells through the use of fluorescence recovery after photobleaching ( P42345 ) analysis and also have characterized the dynamics of the interaction of PR with the mouse mammary tumor virus ( MMTV ) promoter reconstituted into chromatin in vitro . In photobleaching experiments , PR in the presence of the agonist R5020 exhibits rapid exchange with the MMTV promoter in living cells . Two PR antagonists , DB00834 and ZK98299 , have opposite effects on receptor dynamics in vivo . In the presence of DB00834 , PR binds to the promoter and is exchanged more slowly than the agonist-activated receptor . In contrast , PR bound to ZK98299 is not localized to the promoter and exhibits higher mobility in the nucleoplasm than the agonist-bound receptor . Significantly , PR bound to R5020 or DB00834 can recruit the SWI/SNF chromatin remodeling complex to the promoter , but PR activated with ZK98299 can not . Furthermore , we found ligand-specific active displacement of PR from the MMTV promoter during chromatin remodeling in vitro and conclude that the interaction of PR with chromatin is highly dynamic both in vivo and in vitro . We propose that factor displacement during chromatin remodeling is an important component of receptor mobility and that ligand-specific interactions with remodeling complexes can strongly influence receptor nuclear dynamics and rates of exchange with chromatin in living cells . Leptin resistance in vagal afferent neurons inhibits cholecystokinin signaling and satiation in diet induced obese rats . BACKGROUND AND AIMS : The gastrointestinal hormone cholecystokinin ( CCK ) plays an important role in regulating meal size and duration by activating CCK1 receptors on vagal afferent neurons ( Q15025 ) . Leptin enhances CCK signaling in Q15025 via an early growth response 1 ( P18146 ) dependent pathway thereby increasing their sensitivity to CCK . In response to a chronic ingestion of a high fat diet , Q15025 develop leptin resistance and the satiating effects of CCK are reduced . We tested the hypothesis that leptin resistance in Q15025 is responsible for reducing CCK signaling and satiation . RESULTS : Lean Zucker rats sensitive to leptin signaling , significantly reduced their food intake following administration of CCK8S ( 0.22 nmol/kg , i.p. ) , while obese Zucker rats , insensitive to leptin , did not . CCK signaling in Q15025 of obese Zucker rats was reduced , preventing CCK-induced up-regulation of P49146 and down-regulation of melanin concentrating hormone 1 receptor ( Q99705 ) and cannabinoid receptor ( P21554 ) . In Q15025 from diet-induced obese ( DIO ) Sprague Dawley rats , previously shown to become leptin resistant , we demonstrated that the reduction in P18146 expression resulted in decreased sensitivity of Q15025 to CCK and reduced CCK-induced inhibition of food intake . The lowered sensitivity of Q15025 to CCK in DIO rats resulted in a decrease in P28062 expression and increased P21554 and Q99705 expression . These effects coincided with the onset of hyperphagia in DIO rats . CONCLUSIONS : Leptin signaling in Q15025 is required for appropriate CCK signaling and satiation . In response to high fat feeding , the onset of leptin resistance reduces the sensitivity of Q15025 to CCK thus reducing the satiating effects of CCK . Differential in vitro sensitivity to patupilone versus paclitaxel in uterine and ovarian carcinosarcoma cell lines is linked to tubulin-beta-III expression . OBJECTIVE : To compare the in vitro sensitivity/resistance to patupilone versus paclitaxel in uterine and ovarian carcinosarcomas ( CS ) . METHODS : Five primary carcinosarcoma cell lines , two from uterine and three of ovarian origin , were evaluated for growth rate and tested for their in vitro sensitivity/resistance to patupilone versus paclitaxel by MTS assays . To identify potential mechanisms underlying the differential sensitivity/resistance to patupilone , expression levels of β-tubulin III ( Q13509 ) were determined with quantitative-real-time-polymerase-chain-reaction ( q-RT-PCR ) in primary uterine and ovarian CS cell lines and in 26 uterine and 9 ovarian CS fresh-frozen-tissues . RESULTS : No appreciable difference in sensitivity to patupilone versus paclitaxel was noted in ovarian CS cell lines , or when uterine and ovarian CS cell lines were compared in their response to paclitaxel . In contrast , uterine CS cell lines were found to be significantly more sensitive to patupilone than to paclitaxel ( P < 0.002 ) and demostrated lower IC(50s) to patupilone ( range 0.76-0.93nM ) when compared to ovarian CS ( range 1.9-3.4 nM , p < 0.05 ) . Higher levels of Q13509 were detected in uterine CS cell lines and fresh frozen tissues when compared to ovarian CS ( P < 0.05 ) . CONCLUSIONS : Uterine CS cell lines are significantly more sensitive than ovarian CS cell lines to patupilone versus paclitaxel . High expression of Q13509 is associated with sensitivity to patupilone in primary CS cell lines and may act as a genetic marker to predict chemotherapy efficacy . DB03010 may represent a promising drug in the treatment of this subset of rare but highly aggressive gynecological tumors . P06401 -B regulation of insulin-like growth factor-stimulated cell migration in breast cancer cells via insulin receptor substrate-2 . Progesterone action contributes to the signaling of many growth factor pathways relevant to breast cancer tumor biology , including the insulin-like growth factor ( IGF ) system . Previous work has shown that insulin receptor substrate-2 ( Q9Y4H2 ) but not P35568 levels were regulated by progestin in progesterone receptor-B ( PR-B ) isoform expressing MCF-7 cells ( C4-12 PR-B ) . Furthermore , type 1 IGF receptor ( P08069 ) signaling via Q9Y4H2 correlated with the increased cell migration observed in a number of breast cancer cell lines . Consequently , in this study , we examined whether the elevation of Q9Y4H2 protein induced by progestin was sufficient to promote P05019 -stimulated cell motility . Treatment of C4-12 PR-B cells with progestin shifted the balance of phosphorylation from P35568 to Q9Y4H2 in response to P05019 . This shift in Q9Y4H2 activation was associated with enhanced migration in C4-12 PR-B cells pretreated with progestin , but had no effect on cell proliferation or survival . Treatment of C4-12 PR-B cells with DB00834 , an antiprogestin , inhibited IGF-induced cell migration . Attenuation of Q9Y4H2 expression using small interfering RNA resulted in decreased IGF-stimulated motility . In addition , Q9Y4H2 knockdown resulted in an abrogation of P31749 /Akt phosphorylation but not mitogen-activated protein kinase . Consequently , LY294002 , a phosphoinositide-3-kinase inhibitor , abolished IGF-induced cell motility in progestin-treated C4-12 PR-B cells . These data show a role for the PR in functionally promoting growth factor signaling , showing that levels of P41252 proteins can determine IGF-mediated biology , PR-B signaling regulates Q9Y4H2 expression , and that Q9Y4H2 can mediate IGF-induced cell migration via phosphoinositide-3-kinase in breast cancer cells . DB06155 inhibits human colon cancer cell growth and reduces the formation of precancerous lesions in the mouse colon . The selective P21554 receptor antagonist rimonabant ( SR141716 ) was shown to perform a number of biological effects in several pathological conditions . Emerging findings demonstrate that rimonabant exerts antitumor action in thyroid tumors and breast cancer cells . In our study , human colorectal cancer cells ( DLD-1 , CaCo-2 and SW620 ) were treated with rimonabant and analyzed for markers of cell proliferation , cell viability and cell cycle progression . DB06155 significantly reduced cell growth and induced cell death . In addition , rimonabant was able to alter cell cycle distribution in all the cell lines tested . Particularly , rimonabant produced a G2/M cell cycle arrest in DLD-1 cells without inducing apoptosis or necrosis . The G2/M phase arrest was characterized by a parallel enhancement of the number of mitoses associated to elevated DNA double strand breaks and chromosome misjoining events , hallmarks of mitotic catastrophe . Protein expression analyses of P12004 B1 , P09874 , Aurora B and phosphorylated p38/MAPK and Chk1 demonstrated that rimonabant-induced mitotic catastrophe is mediated by interfering with the spindle assembly checkpoint and the DNA damage checkpoint . Moreover , in the mouse model of azoxymethane-induced colon carcinogenesis , rimonabant significantly decreased aberrant crypt foci ( Q9NQ94 ) formation , which precedes colorectal cancer . Our findings suggest that rimonabant is able to inhibit colorectal cancer cell growth at different stages of colon cancer pathogenesis inducing mitotic catastrophe in vitro . Reduction of human monocytic cell neurotoxicity and cytokine secretion by ligands of the cannabinoid-type CB2 receptor . 1 Two cannabinoid receptors , P21554 and CB2 , have been identified . The P21554 receptor is preferentially expressed in brain , and the CB2 receptor in cells of leukocyte lineage . We identified the mRNA for the P21554 receptor in human neuroblastoma SH-SY5Y cells , and the mRNA and protein for the CB2 receptor in human microglia and THP-1 cells . 2 Delta(9)-and Delta(8)-tetrahydrocannabinol ( THC ) were toxic when added directly to SH-SY5Y neuroblastoma cells . The toxicity of Delta(9)- THC was inhibited by the P21554 receptor antagonist SR141716A but not by the CB2 receptor antagonist SR144528 . The endogenous ligand anandamide was also toxic , and this toxicity was enhanced by inhibitors of its enzymatic hydrolysis . 3 The selective CB2 receptor ligands JWH-015 and indomethacin morpholinylamide ( BML-190 ) , when added to THP-1 cells before stimulation with lipopolysaccharide ( LPS ) and P01579 , reduced the toxicity of their culture supernatants to SH-SY5Y cells . JWH-015 was more effective against neurotoxicity of human microglia than THP-1 cells . The antineurotoxic activity of JWH-015 was blocked by the selective CB2 receptor antagonist SR144528 , but not by the P21554 receptor antagonist SR141716A . This activity of JWH-015 was synergistic with that of the P09917 ( 5- P28300 ) inhibitor REV 5901 . 4 Cannabinoids inhibited secretion of IL-1beta and tumor necrosis factor-alpha ( P01375 ) by stimulated THP-1 cells , but these effects could not be directly correlated with their antineurotoxic activity . 5 Specific CB2 receptor ligands could be useful anti-inflammatory agents , while avoiding the neurotoxic and psychoactive effects of P21554 receptor ligands such as Delta(9)-THC .

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Design of novel potent antihyperlipidemic agents with antioxidant/anti-inflammatory properties : exploiting phenothiazine 's strong antioxidant activity . Because atherosclerosis is an inflammatory process involving a series of pathological events such as dyslipidemia , oxidative stress , and blood clotting mechanisms , we hereby report the synthesis and evaluation of novel compounds in which antioxidant , anti-inflammatory , and squalene synthase ( P37268 ) inhibitory/hypolipidemic activities are combined in simple molecules through design . The coupling of two different pharmacophores afforded compounds 1-12 , whose biological profile was markedly improved compared to those of parent lead structures ( i.e. , the hypolipidemic 2-hydroxy-2-aryl-(benzo)oxa ( or thia ) zine and the antioxidant phenothiazine ) . Most derivatives strongly inhibited in vitro microsomal lipid and LDL peroxidation , exhibiting potent free-radical scavenging activity . They further significantly inhibited P37268 activity and showed remarkable antidyslipidemic activity in vivo in animal models of acute and high-fat-induced hyperlipidemia . Finally , several compounds showed anti-inflammatory activity in vitro , inhibiting cycloxygenase ( P23219 /2 ) activity . The multimodal properties of the new compounds and especially their combined antioxidant/ P37268 / P36551 inhibitory activity render them interesting lead compounds for further evaluation against atherosclerosis . DB09078 : first global approval . DB09078 ( Lenvima™ ) is a multitargeted receptor kinase inhibitor that inhibits the kinase activities of vascular endothelial-derived growth factor receptors 1 , 2 and 3 , fibroblast growth factor receptors 1 , 2 , 3 and 4 , platelet-derived growth factor receptor α , P07949 and P10721 . In addition to their role in normal cellular function , these kinases have been implicated in pathogenic angiogenesis , tumour growth and cancer progression . DB09078 is being developed by Eisai Co . Ltd for the treatment of solid tumours , primarily for differentiated thyroid cancer , and other malignancies . A capsule formulation of the drug has received approval in the USA for use in locally recurrent or metastatic , progressive , radioactive iodine-refractory differentiated thyroid cancer . DB09078 is in pre-registration for this indication in the EU , Australia , Brazil , Canada , Japan , South Korea , Russia , Singapore and Switzerland , and is in phase 3 development in Argentina , Chile and Thailand . DB09078 has orphan designation in the EU and Japan for use in differentiated thyroid cancer . In addition , an ongoing global , phase 3 trial is evaluating the use of lenvatinib as first-line treatment in unresectable hepatocellular carcinoma . This article summarizes the milestones in the development of lenvatinib leading to this first approval in locally recurrent or metastatic , progressive , radioactive iodine-refractory differentiated thyroid cancer . Suppression of proliferation of two independent P21359 malignant peripheral nerve sheath tumor cell lines by the pan-ErbB inhibitor DB05424 . Neurofibromatosis Type 1 ( P21359 ) is characterized by the abnormal proliferation of neuroectodermal tissues and the development of certain tumors , particularly neurofibromas , which may progress into malignant peripheral nerve sheath tumors ( MPNSTs ) . Effective pharmacological therapy for the treatment of P21359 tumors is currently unavailable and the prognosis for patients with MPNSTs is poor . Loss of neurofibromin correlates with increased expression of the epidermal growth factor receptor ( P00533 ) and ErbB2 tyrosine kinases and these kinases have been shown to promote P21359 tumor-associated pathologies in vivo . We show here that while P21359 MPNST cells have higher P00533 expression levels and are more sensitive to P01133 when compared to a non- P21359 MPNST cell line , the ability of the P00533 inhibitor gefitinib to selectively inhibit P21359 MPNST cell proliferation is marginal . We also show that P21359 MPNST proliferation correlates with activated ErbB2 and can be suppressed by nanomolar concentrations of the pan-ErbB inhibitor DB05424 ( canertinib ) . Consequently , targeting both P00533 and ErbB2 may prove an effective strategy for suppressing P21359 MPNST tumor growth in vivo . Farnesyl diphosphate synthase : the art of compromise between substrate selectivity and stereoselectivity . Farnesyl diphosphate ( FPP ) synthase catalyzes the consecutive head-to-tail condensations of isopentenyl diphosphate ( IPP , P01031 ) with dimethylallyl diphosphate ( DMAPP , P01031 ) and geranyl diphosphate ( GPP , Q99622 ) to give ( E,E ) -FPP ( C15 ) . The enzyme belongs to a genetically distinct family of chain elongation enzymes that install E-double bonds during each addition of a five-carbon isoprene unit . Analysis of the Q99622 and C15 products from incubations with avian P14324 reveals that small amounts of neryl diphosphate ( Z- Q99622 ) and ( Z,E ) -FPP are formed along with the E-isomers during the P01031 --> Q99622 and Q99622 --> C15 reactions . Similar results were obtained for P14324 from Escherichia coli , Artemisia tridentata ( sage brush ) , Pyrococcus furiosus , and Methanobacter thermautotrophicus and for GPP and FPP synthesized in vivo by E. coli P14324 . When ( R ) -[2-2H]IPP was a substrate for chain elongation , no deuterium was found in the chain elongation products . In contrast , the deuterium in ( S ) -[2-2H]IPP was incorporated into all of the products . Thus , the pro-R hydrogen at P06681 of IPP is lost when the E- and Z-double bond isomers are formed . The synthesis of Z-double bond isomers by P14324 during chain elongation is unexpected for a highly evolved enzyme and probably reflects a compromise between optimizing double bond stereoselectivity and the need to exclude DMAPP from the IPP binding site . Novel immunosuppression : small molecules and biologics . Kidney transplantation today has excellent short-term outcomes that have paralleled the use of new immunosuppressive agents introduced in the 1990s . In addition to reducing acute rejection , the goals for developing new agents is to improve long-term outcome , minimize nephrotoxicity , and reduce infectious , cardiovascular , and malignancy-related complications . Novel small molecules and biological agents currently in clinical development may help to minimize the use of calcineurin inhibitors and steroids . These small molecules include FTY720 , a sphingosine phosphate-receptor modulator , FK778 , an inhibitor of pyrimidine synthesis , CP-690550 , a P52333 inhibitor , and AEB-071 , a protein kinase C inhibitor . The biological agents include drugs targeting interleukin-15 , anti- P25942 , belatacept ( DB06681 ) , a second-generation CTLY4Ig that blocks the interaction between P33681 /86 and P10747 costimulatory pathways , and efalizumab , a humanized anti-LFA1 monoclonal antibody . These new agents currently in preclinical and clinical trials appear promising and may represent the emergence of novel immunosuppressive agents that can deliver immunosuppression without long-term toxicity . Global transcriptome analysis of formalin-fixed prostate cancer specimens identifies biomarkers of disease recurrence . Prostate cancer remains the second leading cause of cancer death in American men and there is an unmet need for biomarkers to identify patients with aggressive disease . In an effort to identify biomarkers of recurrence , we performed global RNA sequencing on 106 formalin-fixed , paraffin-embedded prostatectomy samples from 100 patients at three independent sites , defining a 24-gene signature panel . The 24 genes in this panel function in cell-cycle progression , angiogenesis , hypoxia , apoptosis , PI3K signaling , steroid metabolism , translation , chromatin modification , and transcription . Sixteen genes have been associated with cancer , with five specifically associated with prostate cancer ( P78543 , P17936 , Q96EB6 , P50539 , and P14324 ) . Validation was performed on an independent publicly available dataset of 140 patients , where the new signature panel outperformed markers published previously in terms of predicting biochemical recurrence . Our work also identified differences in gene expression between Gleason pattern 4 + 3 and 3 + 4 tumors , including several genes involved in the epithelial-to-mesenchymal transition and developmental pathways . Overall , this study defines a novel biomarker panel that has the potential to improve the clinical management of prostate cancer . Excision of a lyase-resistant oxidized abasic lesion from DNA . The P06681 '-oxidized abasic lesion ( P06681 -AP ) is produced in DNA that is subjected to oxidative stress . The lesion disrupts replication and gives rise to mutations that are dependent upon the identity of the upstream nucleotide . Ape1 incises P06681 -AP , but the 5'-phosphorylated fragment is not a substrate for the lyase activity of P06746 . Excision of the lesion is achieved by strand displacement synthesis in the presence of flap endonuclease during which P06681 -AP and the 3'-adjacent nucleotide are replaced . The oxidized abasic lesion is also a substrate for the bacterial UvrABC nucleotide excision repair system . These data suggest that the redundant nature of DNA repair systems provides a means for removing a lesion that resists excision by short patch base excision repair . P04141 -1 ( P09603 ) delivers a proatherogenic signal to human macrophages . P09603 / P09603 supports the proliferation and differentiation of monocytes and macrophages . In mice , P09603 also promotes proinflammatory responses in vivo by regulating mature macrophage functions , but little is known about the acute effects of this growth factor on mature human macrophages . Here , we show that in contrast to its effects on mouse bone marrow-derived macrophages , P09603 did not induce expression of urokinase plasminogen activator mRNA , repress expression of apolipoprotein E mRNA , or prime LPS-induced P01375 and P05231 secretion in human monocyte-derived macrophages ( HMDM ) from several independent donors . Instead , we show by expression profiling that P09603 modulates the HMDM transcriptome to favor a proatherogenic environment . P09603 induced expression of the proatherogenic chemokines P02778 /IFN-inducible protein 10 , P13500 , and P80098 but repressed expression of the antiatherogenic chemokine receptor P61073 . P09603 also up-regulated genes encoding enzymes of the cholesterol biosynthetic pathway ( P04035 , P53602 , Q13907 , P14324 , Q14534 , Q16850 , EBP , Q15738 , Q9UBM7 , and Q15392 ) , and expression of P45844 , encoding a cholesterol efflux transporter , was repressed . Consistent with these effects , P09603 increased levels of free cholesterol in HMDM , and the selective P07333 kinase inhibitor GW2580 ablated this response . These data demonstrate that P09603 represents a further link between inflammation and cardiovascular disease and suggest two distinct mechanisms by which P09603 , which is known to be present in atherosclerotic lesions , may contribute to plaque progression . [ DB09043 ( Eperzan ) : a new once-weekly agonist of glucagon-like peptide-1 receptors ] . DB09043 ( Eperzan ) is a new once-weekly agonist of Glucagon-Like Peptide-1 ( P0C6A0 ) receptors that is indicated in the treatment of type 2 diabetes . Two doses are available , 30 mg and 50 mg , to be injected subcutaneously once a week . It has been extensively evaluated in the HARMONY programme of eight large randomised controlled trials that were performed at different stages of type 2 diabetes , in comparison with placebo or an active comparator . The endocrine and metabolic effects of albiglutide are similar to those of other P43220 agonists : stimulation of insulin secretion ( incretin effect ) and inhibition of glucagon secretion , both in a glucose-dependent manner , retardation of gastric emptying and increase of satiety . These effects lead to a reduction in glycated haemoglobin ( HbA(1c) ) levels , combined with a weight reduction . The overall tolerance profile is good . DB09043 is currently reimbursed in Belgium after failure ( HbA(1c) > 7.5 % ) of and in combination with a dual therapy with metformin and a sulfonylurea as well as in combination with a basal insulin ( with or without oral antidiabetic drugs ) . To avoid hypoglycaemia , a reduction in the dose of sulfonylurea or insulin may be recommended . A once-weekly administration should increase patient 's acceptance of injectable therapy and improve compliance . Effects of non-steroidal anti-inflammatory drugs on cyclo-oxygenase and lipoxygenase activity in whole blood from aspirin-sensitive asthmatics vs healthy donors . 1. Cyclo-oxygenase ( P36551 ) and lipoxygenase ( LO ) share a common substrate , arachidonic acid . DB00945 and related drugs inhibit P36551 activity . In a subset of patients with asthma aspirin induces clinical symptoms associated with increased levels of certain LO products , a phenomenon known as aspirin-sensitive asthma . The pharmacological pathways regulating such responses are not known . 2 . Here P23219 and LO activity were measured respectively by the formation of thromboxane B(2) ( TXB(2) ) or leukotrienes ( LT ) C(4) , D(4) and E(4) in whole blood stimulated with A23187 . P35354 activity was measured by the formation of prostaglandin E(2) ( PGE(2) ) in blood stimulated with lipopolysaccharide ( LPS ) for 18 h . 3 . No differences in the levels of P23219 , P35354 or LO products or the potency of drugs were found in blood from aspirin sensitive vs aspirin tolerant patients . DB00945 , indomethacin and nimesulide inhibited P23219 activity , without altering LO activity . Indomethacin , nimesulide and the P35354 selective inhibitor DB00677 [ 5,5-dimethyl-3-(2-isopropoxy)-4-(4-methanesulfonylphenyl)-2(5H)-furanone ] inhibited P35354 activity . NO-aspirin , like aspirin inhibited P23219 activity in blood from both groups . However , NO-aspirin also reduced LO activity in the blood from both patient groups . Sodium salicylate was an ineffective inhibitor of P23219 , P35354 or LO activity in blood from both aspirin-sensitive and tolerant patients . 4 . Thus , when P36551 activity in the blood of aspirin-sensitive asthmatics is blocked there is no associated increase in LO products . Moreover , NO-aspirin , unlike other NSAIDs tested , inhibited LO activity in the blood from both aspirin sensitive and aspirin tolerant individuals . This suggests that NO-aspirin may be better tolerated than aspirin by aspirin-sensitive asthmatics . The design and development of pegfilgrastim ( PEG-rmetHuG- P04141 , Neulasta ) . Recombinant protein technology produces drugs for human therapy in unprecedented quantity and quality . Research is now focusing on the relationship between pharmacokinetic and pharmacodynamic properties of molecules , with the aim of engineering proteins that possess enhanced therapeutic characteristics in contrast to being used as simple replacements for the natural equivalent . The addition of a polyethylene glycol ( PEG ) moiety to filgrastim ( rmetHu- DB00099 , Neupogen ) resulted in the development of pegfilgrastim . DB00019 is a long-acting form of filgrastim that requires only once-per-cycle administration for the management of chemotherapy-induced neutropenia . The covalent attachment of PEG to the N-terminal amine group of the parent molecule was attained using site-directed reductive alkylation . Pegylation increases the size of filgrastim so that it becomes too large for renal clearance . Consequently , neutrophil-mediated clearance predominates in elimination of the drug . This extends the median serum half-life of pegfilgrastim to 42 hours , compared with between 3.5 and 3.8 hours for DB00099 , though in fact the half-life is variable , depending on the absolute neutrophil count , which in turn reflects of the ability of pegfilgrastim to sustain production of those same cells . The clearance of the molecule is thus dominated by a self-regulating mechanism . DB00019 retains the same biological activity as filgrastim , and binds to the same Q99062 , stimulating the proliferation , differentiation and activation of neutrophils . Once-per-chemotherapy cycle administration of pegfilgrastim reduces the duration of severe neutropenia as effectively as daily treatment with filgrastim . In clinical trials , patients receiving pegfilgrastim also had a lower observed incidence of febrile neutropenia than patients receiving filgrastim . Genome-wide analysis of DNA copy number alterations and gene expression in gastric cancer . Genomic copy number aberrations ( CNAs ) are believed to play a major role in the development and progression of human cancers . Although many CNAs have been reported in gastric cancer , their genome-wide transcriptional consequences are poorly understood . In this study , to reveal the impact of CNAs on genome-wide expression in gastric cancer , we analysed 30 cases of gastric cancers for their CNAs by array comparative genomic hybridization ( array CGH ) and 24 of these 30 cases for their expression profiles by oligonucleotide-expression microarray . We found that with the application of laser microdissection , most CNAs were detected at higher frequency than in previous studies . Notably , gain at 20q13 was detected in almost all cases ( 97 % ) , suggesting that this may play an important role in the pathogenesis of gastric cancer . By comparing the array CGH data with expression profiles of the same samples , we showed that both genomic amplification and deletion strongly influence the expression of genes in altered genomic regions . Furthermore , we identified 125 candidate genes , consisting of 114 up-regulated genes located in recurrent regions ( > 10 % ) of amplification and 11 down-regulated genes located in recurrent regions of deletion . Up-regulation of several candidate genes , such as Q99741 , P60059 , Q9BTT0 , Q13895 and P37268 , was confirmed by immunohistochemistry . Interestingly , some candidate genes were localized at genomic loci adjacent to well-known genes such as P00533 , P04626 and Q13485 , and concordantly deregulated by genomic alterations . Based on these results , we propose that our list of candidate genes may contain novel genes involved in the pathogenesis of advanced gastric cancer . Angiotensin converting enzyme inhibitor attenuates oxidative stress-induced endothelial cell apoptosis via p38 Q96HU1 kinase inhibition . BACKGROUND : The effects of angiotensin converting enzyme ( P12821 ) inhibitors on oxidative stress-induced apoptosis of endothelial cells and the intracellular signaling were investigated . METHODS : Cultured endothelial cells derived from a bovine carotid artery were treated with H2O2 or P01375 to induce apoptosis . Apoptosis was evaluated by DNA fragmentation and cell viability , p38 Q96HU1 kinase activity by Western blotting , and oxidative stress by formation of 8-isoprostane . The effects of P12821 inhibitors were examined by adding them into the medium throughout the experiments . RESULTS : Apoptosis was attenuated by P12821 inhibitors , temocapril and captopril , in a dose-dependent manner ( 1-100 micromol/l ) . H2O2 ( 0.2 mmol/l for 1.5 h ) or P01375 ( 10 ng/ml for 72 h ) treatment stimulated the activities of p38 Q96HU1 kinase . DB08836 and captopril decreased the activity of p38 Q96HU1 kinase as well as 8-isoprostane formation induced by H2O2 . A p38 Q96HU1 kinase inhibitor , SB203580 , partially inhibited the effect of temocapril on apoptosis . CONCLUSIONS : These results suggest that P12821 inhibitors protect endothelial cells from oxidative stress-induced apoptosis , and that p38 Q96HU1 kinase plays a critical role in the process . Drug-induced activation of SREBP-controlled lipogenic gene expression in CNS-related cell lines : marked differences between various antipsychotic drugs . BACKGROUND : The etiology of schizophrenia is unknown , but neurodevelopmental disturbances , myelin- and oligodendrocyte abnormalities and synaptic dysfunction have been suggested as pathophysiological factors in this severe psychiatric disorder . DB04540 is an essential component of myelin and has proved important for synapse formation . Recently , we demonstrated that the antipsychotic drugs clozapine and haloperidol stimulate lipogenic gene expression in cultured glioma cells through activation of the sterol regulatory element-binding protein ( SREBP ) transcription factors . We here compare the action of chlorpromazine , haloperidol , clozapine , olanzapine , risperidone and ziprasidone on SREBP activation and SREBP-controlled gene expression ( ACAT2 , P04035 , Q01581 , P14324 , O75845 , Q9UBM7 , P01130 , P49327 and SCD1 ) in four CNS-relevant human cell lines . RESULTS : There were marked differences in the ability of the antipsychotic drugs to activate the expression of SREBP target genes , with clozapine and chlorpromazine as the most potent stimulators in a context of therapeutically relevant concentrations . Glial-like cells ( GaMg glioma and CCF-STTG1 astrocytoma cell lines ) displayed more pronounced drug-induced SREBP activation compared to the response in Q9UL51 human cortical neurons and SH-SY5Y neuroblastoma cells , indicating that antipsychotic-induced activation of lipogenesis is most prominent in glial cells . CONCLUSION : Our present data show a marked variation in the ability of different antipsychotics to induce SREBP-controlled transcriptional activation of lipogenesis in cultured human CNS-relevant cells . We propose that this effect could be relevant for the therapeutic efficacy of some antipsychotic drugs . Effects of DNA polymerase inhibitors on replicative and repair DNA synthesis in ultraviolet-irradiated HeLa cells . Aphidicolin specifically inhibits eukaryotic DNA polymerase alpha , while 2',3'-dideoxythymidine 5'-triphosphate ( d2TTP ) inhibits P06746 and gamma but not alpha . DB00987 5'-triphosphate ( araCTP ) inhibits both DNA polymerase alpha and beta although to a different extent . Here we measured the effects of these inhibitors on repair DNA synthesis of U.V.-irradiated HeLa cells by two different methods . Firstly , aphidicolin , 1-beta-D-arabinofuranosylcytosine ( araC , a precursor of araCTP ) and 2',3'-dideoxythimidine ( d2Thd , a precursor of d2TTP ) were added directly to the culture medium . In this case , aphidicolin and araC strongly inhibited replicative DNA synthesis of HeLa cells , and they also inhibited repair synthesis after U.V.-irradiation but to a much lesser extent . In contrast , high concentrations of d2Thd inhibited repair DNA synthesis to a higher extent than replicative DNA synthesis . Secondly , the active form of inhibitor , d2TTP , was microinjection directly into cytoplasm or nuclei or U.V.-irradiated HeLa cells . Microinjection of d2TTP effectively inhibited repair synthesis . The microinjection of d2TTP , into either cytoplasm or nucleus , strongly inhibited replicative synthesis . These results might indicate that multiple DNA polymerases are involved in repair synthesis as well as in replicative synthesis . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . Comparison of the independent and combined effects of sub-chronic therapy with metformin and a stable P43220 agonist on cognitive function , hippocampal synaptic plasticity and metabolic control in high-fat fed mice . Cognitive dysfunction is more common in individuals with type 2 diabetes ( T2DM ) . Currently , glucagon-like peptide-1 ( P0C6A0 ) and metformin are important therapeutic options for patients with T2DM . However , their potential effects on cognitive function , including underlying mechanisms , are yet to be fully determined . We have compared the individual and combined effects of treatment for 20 days with ( DB00161 (8)) P0C6A0 (GluPAL) , an enzymatically stable P0C6A0 -receptor agonist , and metformin on metabolic control and aspects of learning and memory in high fat fed mice . ( DB00161 (8)) P0C6A0 (GluPAL) treatment for 20 days alone , or in combination with metformin , improved ( p < 0.05 ) the recognition index in high fat mice , indicating enhanced learning and memory . In addition , these mice exhibited a complete reversal of the deleterious effects of prolonged high-fat feeding on long-term potentiation in the hippocampal P00915 region . This was linked to reduced hippocampal levels of 8-oxoguanine ( p < 0.01 ) and glial fibriallary acidic protein ( p < 0.001 ) , indicating decreased oxidative stress and inflammation ; respectively . Expression of fundamental hippocampal genes including P42345 , P15692 , Q16620 and Q96EB6 was also increased significantly ( p < 0.001 ) by all treatments . ( DB00161 (8)) P0C6A0 (GluPAL) monotherapy , or in combination with metformin , reduced circulating glucose ( p < 0.05 ) and increased insulin ( p < 0.05 to p < 0.01 ) concentrations , as well as improving glucose tolerance ( p < 0.001 ) and glucose-stimulated insulin secretion ( p < 0.05 to p < 0.01 ) . P01308 sensitivity and measurements of energy regulation and metabolic rate were not altered . These studies highlight the neuroprotective properties of ( DB00161 (8)) P0C6A0 (GluPAL) , alone and in combination with metformin , in T2DM . Transgenic Panax ginseng inhibits the production of P01375 , P05231 , and P10145 as well as P35354 expression in human mast cells . The most well-known medicinal plant , Panax ginseng ( P. ginseng ) , contains various phytosterols and bioactive triterpene saponins ( ginsenosides ) . P37268 is a key regulatory enzyme for triterpene biosynthesis and overexpression of the squalene synthase confers the hyper-production of triterpene saponins to form transgenic ginseng . In this study , we have investigated whether and how transgenic P. ginseng modulates an inflammatory reaction in a stimulated human mast cell line , HMC-1 . It was found that transgenic P. ginseng inhibited the production of tumor necrosis factor ( P01375 ) -alpha , interleukin ( IL ) -6 , P10145 , and the expression of cyclooxygenase-2 in phorbol 12-myristate 13-acetate ( PMA ) plus calcium ionophore A23187 ( PMACI ) -stimulated HMC-1 . Additionally , we have shown that transgenic P. ginseng suppressed the intracellular calcium level induced by PMACI . These results provide new insights into the pharmacological actions of transgenic P. ginseng as a potential molecule for use in therapy in mast cell-mediated inflammatory diseases . A novel bisphosphonate inhibitor of squalene synthase combined with a statin or a nitrogenous bisphosphonate in vitro . Statins and nitrogenous bisphosphonates ( NBP ) inhibit 3-hydroxy-3-methylglutaryl-coenzyme-A reductase ( P04035 ) and farnesyl diphosphate synthase ( P14324 ) , respectively , leading to depletion of farnesyl diphosphate ( FPP ) and disruption of protein prenylation . P37268 ( P37268 ) utilizes FPP in the first committed step from the mevalonate pathway toward cholesterol biosynthesis . Herein , we have identified novel bisphosphonates as potent and specific inhibitors of P37268 , including the tetrasodium salt of 9-biphenyl-4,8-dimethyl-nona-3,7-dienyl-1,1-bisphosphonic acid ( compound 5 ) . Compound 5 reduced cholesterol biosynthesis and lead to a substantial intracellular accumulation of FPP without reducing cell viability in HepG2 cells . At high concentrations , lovastatin and zoledronate impaired protein prenylation and decreased cell viability , which limits their potential use for cholesterol depletion . When combined with lovastatin , compound 5 prevented lovastatin-induced FPP depletion and impairment of protein farnesylation . Compound 5 in combination with the NBP zoledronate completely prevented zoledronate-induced impairment of both protein farnesylation and geranylgeranylation . Cotreatment of cells with compound 5 and either lovastatin or zoledronate was able to significantly prevent the reduction of cell viability caused by lovastatin or zoledronate alone . The combination of an P37268 inhibitor with an P04035 or P14324 inhibitor provides a rational approach for reducing cholesterol synthesis while preventing nonsterol isoprenoid depletion . Four-week administration of nimesulide , a cyclooxygenase-2 inhibitor , improves endothelial dysfunction in the hindlimb vasculature of streptozotocin-induced diabetic rats . The aim of this study was to examine the effect of chronic administration of nimesulide , a cyclooxygenase-2 inhibitor , on endothelial dysfunction in streptozotocin-induced diabetic rats . Three groups of Sprague-Dawley rats ( 300-350 g , n=6 ) were used . The first group served as normoglycemic control and the second and third groups were rendered diabetic by an intraperitoneal injection of streptozotocin ( 60 mg/kg ) . The third group received the selective P35354 inhibitor , nimesulide ( 20 mg/kg/day ) , orally by gavage for 4 weeks while the second group received only drinking water and served as diabetic control . At the end of the treatment period , the rats were anesthetized with urethane ( 1.2 g/kg ) and mean arterial pressure , heart rate and hindlimb blood flow were monitored . This was followed by the injection of acetylcholine ( endothelium-dependent vasodilator , 0.1-0.8 microg/kg ) and sodium nitroprusside ( endothelium-independent vasodilator 1-4 microg/kg ) . Mean arterial pressure was significantly reduced and hindlimb vascular conductance was not significantly affected in the control diabetic group when compared to the normoglycemic control group . DB04743 treatment did not cause any significant change in any of the measured hemodynamic parameters . DB03128 and sodium nitroprusside induced dose-dependent increases in hindlimb vascular conductance in control normoglycemic rats which were attenuated in diabetic control rats . DB04743 reversed the attenuation of acetylcholine-induced increase in hindlimb vascular conductance . In conclusion , chronic administration of the selective P35354 inhibitor , nimesulide improved endothelial dysfunction in the hindlimb vasculature of streptozotocin induced diabetic rats . This suggests that P35354 products might be involved in the pathogenesis of endothelial dysfunction in streptozotocin-induced diabetic rats . Leukocyte P29965 deficiency affects the CD25(+) P01730 T cell population but does not affect atherosclerosis . Inhibition of P25942 - P29965 interactions results in a reduction of innate regulatory T cells ( Tregs ) in P25942 (-/-) mice and induces a stable plaque phenotype in atherosclerosis-prone mouse strains . Here we investigated the effects of leukocyte P29965 on the Treg population and on atherosclerosis . P01130 (-/-) mice were reconstituted with wild-type or P29965 (-/-) bone marrow ( BM ) . These BM chimeras were analysed by flow cytometry for the presence of innate Tregs ( CD45RB(low) CD25(+) P01730 ) in lymphoid organs and peripheral blood . As in P25942 (-/-) mice , the CD45RB(high):CD45RB(low) P01730 T cell ratio significantly increased and the CD25(+) P01730 (+) subpopulation significantly decreased in P01130 (-/-) mice receiving P29965 (-/-) BM compared to P01130 (-/-) mice receiving wild-type BM . However , atherosclerotic plaque progression and plaque phenotype did not change in P01130 (-/-) mice reconstituted with P29965 (-/-) BM . In conclusion , the present study shows that P25942 - P29965 interactions on leukocytes are essential for the size of the CD45RB(low) CD25(+) P01730 Treg subpopulation . Nevertheless , P29965 deficiency on hemopoietic cells did not affect atherosclerosis , implying that P29965 expressing leukocytes alone are not responsible for the stable plaque phenotype observed after total P29965 blockade . A new gene mapping resource : interspecies hybrids between Père David 's deer ( Elaphurus davidianus ) and red deer ( Cervus elaphus ) . Three male F1 hybrids between Père David 's deer and red deer were mated to red deer to produce 143 backcross calves . The pedigrees are a rare example of a fertile hybrid between evolutionarily divergent species . We examined the use of these families for genetic mapping of evolutionarily conserved ( Type I ) loci by testing for genetic linkage between five species-specific protein variants and 12 conserved DNA probes . Two probes were homologous , and the remainder syntenic , to the protein coding loci in cattle or humans . Using six restriction enzymes , each DNA probe detected one or more restriction fragments specific to Père David 's deer . Linkage analyses among the species-specific variants placed the loci into four linkage groups within which linkage between adjacent loci and gene order was supported by a LOD > 3 . The linkage groups were ( P02790 , P68871 ) - P01225 - P11117 , P00338 - P06127 - P01344 , P12645 - ( GC , ALB ) - ( P10721 , P16234 ) and P01130 - P01024 - P05230 . Southern and protein analysis of P00338 and ALB provided identical segregation data . These linkage groups were consistent with the cattle gene map and provide new information for comparing the gene maps of ruminants , humans and mice . The deer hybrids are an important new resource that can contribute to the comparative analysis of the mammalian genome .

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MEK inhibition in non-small cell lung cancer . P01116 mutations are the most common mutations in non-small cell lung cancer ( NSCLC ) with adenocarcinoma histology . P01116 mutations result in the activation of the RAF-MEK- P29323 pathway , and agents that target RAF-MEK- P29323 pathways have been investigated in P01116 mutant NSCLC . The two agents furthest in development are selumetinib and trametinib . DB08911 has greater binding for the Q02750 /2 allosteric site , and generally has superior pharmacokinetics . A randomized phase II trial of docetaxel with and without selumetinib revealed that the combination resulted numerically superior overall survival , and a statistically significant improvement in progression-free survival and objective response rate . However , a concerning rate of hospital admission , grade 3 or 4 neutropenia , and febrile neutropenia was observed with the combination . Trials have investigated MEK inhibitors as single agents and in combination with erlotinib , and the data do not support the further development . The activity of MEK inhibitors appears to be similar in patients with P01116 mutant and wild-type NSCLC suggesting P01116 mutation status is not a reliable biomarker for efficacy . It is possible that mutations of genes in addition to P01116 mutations impact the activity of MEK inhibitors , or specific subsets of P01116 mutations may be resistant or susceptible to MEK inhibition . Other potential explanations are gene amplifications , alternative RNA splicing of genes resulting in activation of their protein products , and deregulation of noncoding RNAs and consequent altered protein expression . P40933 affects serotonin system and exerts antidepressive effects through IL15Rα receptor . Contrary to the reduction of depressive-like behavior observed in several strains of cytokine receptor knockout mice , mice lacking the specific receptor for interleukin ( IL ) -15 showed increased immobility in tail suspension and modified forced swimming tests . There was also a reduction in social interactions . The hippocampus of the IL15Rα knockout mice had decreased mRNA for 5-HT(1A) , increased mRNA for 5-HT(2C) , and region-specific changes of serotonin reuptake transporter ( P31645 ) immunoreactivity . DB00472 ( the classic antidepressant DB00472 , which inhibits 5-HT(2C) and P31645 ) reduced the immobility of the IL15Rα knockout mice in comparison with their pretreatment baseline . Together with the unchanged performance of the IL15Rα knockout mice on the rotarod , this response to fluoxetine indicates that the immobility reflects depression . Wildtype mice responded to P40933 treatment with improvement of immobility induced by forced swimming , whereas the knockout mice failed to respond . Thus , the cognate P40933 receptor is necessary for the antidepressive activity of P40933 . In ex vivo studies , P40933 decreased synaptosomal uptake of 5-HT , and modulated the expression of 5-HT(2C) and P31645 in cultured neurons in a dose- and time-dependent manner . Thus , the effect of P40933 on serotonin transmission may underlie the depressive-like behavior of IL15Rα knockout mice . We speculate that P40933 is essential to maintain neurochemical homeostasis and thereby plays a role in preventing neuropsychiatric symptoms . Distribution of polyamines and their biosynthetic enzymes in intestinal adaptation . P11926 ( ODC ) and the polyamines have been shown to be important for growth processes in the intestinal mucosa . The highest activity of ODC is found in the differentiated , nonproliferating villus-tip cells rather than in the rapidly proliferating undifferentiated crypt cells . During poststarvation refeeding and lactation , we now show that increases in ODC activity paralleled the time course of mucosal hyperplasia and thymidine incorporation . Increases in ODC ( threefold ) were similar in villus and crypt cells , and the villus-crypt gradient of decreasing ODC activity ( 40:1 ) was maintained . The activity of the other polyamine biosynthetic enzyme , S-adenosylmethionine decarboxylase ( P18827 ) , was highest in the crypt cells in the basal state and increased throughout the entire villus-crypt axis during refeeding and lactation , preserving a villus-crypt gradient opposite to that of ODC . During hyperplasia , all three polyamines increased . DB01917 was highest in the villus-tip cells , paralleling ODC activity , whereas spermidine and spermine were highest in the crypt cells and paralleled the distribution of P18827 activity . Thus P18827 activity and spermidine and spermine content may play a more important role than ODC and putrescine in regulation of intestinal mucosal proliferation . It is also possible that the threefold increases in the low levels of ODC in the crypt cells are adequate to trigger cell proliferation , whereas the higher ODC levels in villus cells may represent an association with the differentiation of the enterocytes . Mechanism of inhibition of the P42262 AMPA receptor channel opening by talampanel and its enantiomer : the stereochemistry of the 4-methyl group on the diazepine ring of 2,3-benzodiazepine derivatives . Stereoselectivity of 2,3-benzodiazepine compounds provides a unique way for the design of stereoisomers as more selective and more potent inhibitors as drug candidates for treatment of the neurological diseases involving excessive activity of AMPA receptors . Here we investigate a pair of enantiomers known as DB04982 and its ( + ) counterpart about their mechanism of inhibition and selectivity toward four AMPA receptor subunits or P42261 -4 . We show that DB04982 is the eutomer with the endismic ratio being 14 for the closed-channel and 10 for the open-channel state of P42262 . Kinetic evidence supports that DB04982 is a noncompetitive inhibitor and it binds to the same site for those 2,3-benzodiazepine compounds with the C-4 methyl group on the diazepine ring . This site , which we term as the " M " site , recognizes preferentially those 2,3-benzodiazepine compounds with the C-4 methyl group being in the R configuration , as in the chemical structure of DB04982 . Given that DB04982 inhibits P42261 and P42262 , but is virtually ineffective on the P42263 and P48058 AMPA receptor subunits , we hypothesize that the " M " site(s) on P42261 and P42262 to which DB04982 binds is different from that on P42263 and P48058 . If the molecular properties of the AMPA receptors and DB04982 are used for selecting an inhibitor as a single drug candidate for controlling the activity of all AMPA receptors in vivo , DB04982 is not ideal . Our results further suggest that addition of longer acyl groups to the N-3 position should produce more potent 2,3-benzodiazepine inhibitors for the " M " site . DB00472 -induced proliferation and differentiation of neural progenitor cells isolated from rat postnatal cerebellum . Previous studies have shown that the serotonin-reuptake inhibitor ( SSRI ) fluoxetine affects neural progenitors derived from postnatal cerebellum or adult hippocampus and stimulates their proliferation . In the human cerebellum , the proliferation of cerebellar granule cells ( CGC ) continues until the 11th postnatal month and could be influenced in infants by breastfeeding-delivered SSRIs . Current information about fluoxetine effects on postnatal cerebellar neural progenitors is limited . Here we report the characterization of fluoxetine actions on rat postnatal cerebellar neural progenitors . RT-PCR and immunostaining revealed the expression of serotonin transporter ( P31645 ) , 5HT(1A) receptors , tryptophan hydroxylase ( P17752 ) , and serotonin ( 5HT ) . Protracted in vitro fluoxetine treatment increased cell proliferation and differentiation . The proliferative effects of fluoxetine , 5HT , and the selective agonist of 5HT(1A) receptors trans-8-hydroxy-2-(N-n-propyl-N-3'-iodo-2'-propenyl)aminotetralin ( 8-OH-PIPAT ) were abolished by the selective antagonist of 5HT(1A) receptors , N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridinyl)cyclohexanecarboxamide trihydrochloride ( WAY-100635 ) . Furthermore , fluoxetine-induced activation of both the DB02527 -response element-binding ( CREB ) protein and extracellular signal-regulated protein kinases ( P27361 /2 ) , which was abolished by the selective inhibitor of Q96HU1 kinase kinase ( MEK ) 1,4-diamino-2,3-dicyano-1,4-bis(2-aminophenylthio)butadiene ( U0126 ) , and increased cyclin D1 expression . All these effects were prevented by WAY-100635 . Collectively , our results demonstrate that rat postnatal cerebellum contains neural progenitors capable of proliferating and differentiating in response to fluoxetine exposure , possibly through the activation of 5HT(1A) receptors . The relevance of these findings for possible SSRI effects on the developing postnatal/infant human cerebellum should be explored . [ Thrombotic complications following the treatment of multiple myeloma with new agents ] . Patients with multiple myeloma ( MM ) are at an increased risk of venous and arterial thrombosis . The risk factors and pathomechanisms for thrombotic complications in multiple myeloma patients can be divided into the disease-related and treatment-specific risk factors . With the introduction of novel therapies , including talidomide , lenalidomide and bortezomib , the outcomes of the patients with newly diagnosed or previously treated multiple myeloma have improved , however the treatment affected the prothrombotic and anticoagulant processes . The risk of venous thromboembolism ( VTE ) in patients receiving immunomodulatory agent-based regimens ( thalidomide or lenalidomide ) , especially when used in combination with high-dose deamethasone- and/or anthracycline-based chemiotherapy is high . The proposed mechanisms for prothrombotic state include changes in P04275 , factor VIII , thrombomodulin , P25116 and P35354 epression , and some abnormalities in transcription factors and genetic risk factors . Moreover , dysregulation of anticoagulation and impairment of fibrinolysis may also contribute to hypercoagulability state . The incidence of VTE in bortezomib-containing regimens is very low . It may be due to inhibitory effect of bortezomib on platelet aggregation and NF-kappa/beta . This article presents the latest outlook upon the pathogenesis of thrombotic complications in multiple myeloma patients undergoing the therapy with new agents . Serotonin increases P27361 /2 phosphorylation in astrocytes by stimulation of P41595 and P28335 receptors . We have previously shown that fluoxetine causes P29323 (1/2) phosphorylation in cultured mouse astrocytes mediated exclusively by stimulation of 5-HT(2B) receptors ( Li et al. , 2008b ) . This raises the question whether this is also the case for serotonin ( 5-HT ) itself . In the present study serotonin was found to induce P29323 (1/2) phosphorylation by stimulation of 5-HT(2B) receptors with high affinity ( EC(50) : 20-30 pM ) , and by stimulation of 5-HT(2C) receptor with low affinity ( EC(50) : 1 microM or higher ) . P29323 (1/2) phosphorylation induced by stimulation of either 5-HT(2B) or 5-HT(2C) receptors was mediated by epidermal growth factor ( P01133 ) receptor transactivation ( Peng et al. , this issue ) , shown by the inhibitory effect of AG1478 , an inhibitor of the P01133 receptor tyrosine kinase , and DB02255 , an inhibitor of Zn-dependent metalloproteinases , and thus of 5-HT(2B) receptor-mediated P01133 receptor agonist release . It is discussed that the high potency of the 5-HT(2B)-mediated effect is consistent with literature data for binding affinity of serotonin to cloned human 5-HT(2B) receptors and with observations of low extracellular concentrations of serotonin in brain , which would allow a demonstrated moderate and modality-dependent increase in specific brain areas to activate 5-HT(2B) receptors . In contrast the relevance of the observed 5-HT(2C) receptors on astrocytes is questioned . The polymorphism IL-1beta T-31C is associated with a longer overall survival in patients with multiple myeloma undergoing auto- P09683 . Proinflammatory cytokines are suspected to play a role in the pathogenesis of multiple myeloma ( MM ) . Therefore , it is possible that inborn genetic variations leading to a modified expression of these cytokines will influence the outcome for these patients . We investigated 348 MM patients undergoing high-dose melphalan treatment followed by Auto- P09683 and examined the influence of single nucleotide polymorphisms ( SNPs ) in genes involved in the inflammatory response . We found that the polymorphism IL-1beta T-31C significantly influenced overall survival ( OS ; P=0.02 ) and that carriers of the variant C-allele had a significantly longer survival than homozygous wild-type allele TT-carriers ( relative risk 0.6 ( 95 % CI=0.5-0.9 ) ; P=0.008 ) . The polymorphisms P05231 G-174C , P22301 C592A , PPARgamma2 Pro(12)Ala , P35354 A-1195G , P35354 T8473C and P19838 ins/del did not influence the OS in this group of patients . Furthermore , homozygous carriers of the variant allele of IL-1beta T-31C were at 1.37-fold ( CI=1.05-1.80 ) increased risk of MM as compared with population-based controls ( P=0.02 ) . Our results indicate that IL-1beta is involved in the pathogenesis of MM . DB00087 for the prevention and treatment of graft-versus-host disease . DB00087 is a humanized monoclonal antibody against the P31358 antigen , which is expressed on the surface of various hematopoietic cells such as B and T lymphocytes , and has been widely used for preventing acute graft-versus-host disease ( GVHD ) in allogeneic stem cell transplantation ( P09683 ) . Administration of 100 mg alemtuzumab before transplantation has resulted in a low incidence of acute GVHD in HLA-matched and mismatched transplantation from either related or unrelated donors . However , because alemtuzumab could remain in the blood at the lympholytic level 1-2 months after transplantation , immune reconstitution was substantially delayed , leading to a high incidence of viral infection and relapse . A dose reduction of alemtuzumab was attempted in a reduced-intensity conditioning setting to facilitate immune reconstitution , and this resulted in earlier immune reconstitution , but the clinical benefits were unclear . The dose of alemtuzumab and the timing of its administration should be optimized to maximize the benefit of acute GVHD suppression and minimize the risk of infection and relapse . Another strategy to facilitate immune reconstitution and augment anti-tumor effects is donor cell infusion of T and NK cells . Although there is accumulating evidence regarding the use of alemtuzumab for acute GVHD prevention , information on the salvage treatment for steroid-refractory acute and chronic GVHD is still limited . Gene therapy-mediated delivery of targeted cytotoxins for glioma therapeutics . Restricting the cytotoxicity of anticancer agents by targeting receptors exclusively expressed on tumor cells is critical when treating infiltrative brain tumors such as glioblastoma multiforme ( GBM ) . GBMs express an P35225 receptor ( IL13Rα2 ) that differs from the physiological P24394 /IL13R receptor . We developed a regulatable adenoviral vector ( Ad.mhIL-4.TRE.mhIL-13-PE ) encoding a mutated human P35225 fused to Pseudomonas exotoxin ( mhIL-13-PE ) that specifically binds to IL13Rα2 to provide sustained expression , effective anti-GBM cytotoxicity , and minimal neurotoxicity . The therapeutic Ad also encodes mutated human P05112 that binds to the physiological P24394 /IL13R without interacting with IL13Rα2 , thus inhibiting potential binding of mhIL-13-PE to normal brain cells . Using intracranial GBM xenografts and syngeneic mouse models , we tested the Ad.mhIL-4.TRE.mhIL-13-PE and two protein formulations , hIL-13-PE used in clinical trials ( DB05305 ) and a second-generation mhIL-13-PE . DB05305 doubled median survival without eliciting long-term survival and caused severe neurotoxicity ; mhIL-13-PE led to ∼40 % long-term survival , eliciting severe neurological toxicity at the high dose tested . In contrast , Ad-mediated delivery of mhIL-13-PE led to tumor regression and long-term survival in over 70 % of the animals , without causing apparent neurotoxicity . Although DB05305 was originally developed to target GBM , when tested in a phase III trial it failed to achieve clinical endpoints and revealed neurotoxicity . Limitations of DB05305 include its short half-life , which demanded frequent or continued administration , and binding to P24394 /IL13R , present in normal brain cells . These shortcomings were overcome by our therapeutic Ad , thus representing a significant advance in the development of targeted therapeutics for GBM . Transcriptional profiles during the differentiation and maturation of monocyte-derived dendritic cells , analyzed using focused microarrays . Dendritic cells ( DC ) are professional antigen-presenting cells capable of initiating primary immune responses . They have been intensively studied and are used in both basic immunology research and clinical immunotherapy . However , the genetic pathways leading to DC differentiation and maturation remain poorly understood . Using focused microarrays with oligonucletotide probes for 120 genes encoding co-stimulatory molecules , chemokines , chemokine receptors , cytokines , cytokine receptors , TLRs , and several other related molecules , we analyzed the kinetics of gene expression for the overall differentiation process of monocytes into mature DC . In parallel , we compared the transcriptional profiles in DC maturation in the presence of LPS , P01375 or trimeric P29965 . We found similar transcriptional profiles for early immature DC and immature DC , respectively generated by culturing monocytes with GM- P04141 and P05112 for three or six days . We identified sets of common and stimuli-specific genes , the expression of which changed following stimulation with LPS , P01375 or P29965 . A dynamic analysis of the entire DC differentiation and maturation process showed that some important inflammatory and constitutive chemokines are transcribed in both immature and mature DC . The correlative expression kinetics of the gene pairs P14778 / P27930 , P40933 / Q13261 , Q9NNX6 / P13598 and Q9NNX6 / P32942 imply that they all play crucial roles in mediating DC functions . Thus , our analysis with focused microarrays shed light on the transcriptional kinetics of DC differentiation and maturation , and this method may also prove useful for identifying novel marker genes involved in DC functions . 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 . Current situation of DB01269 , DB05101 , Nimotuzumab and Zalutumumab . P00533 overexpression usually correlates with a more advanced disease stage , a poorer prognosis and a worse chemotherapy response . P00533 expression increase has been observed in many tumours . For all the aforementioned reasons , P00533 inhibition can be considered an attractive approach in cancer treatment . One strategy has been receptor inhibition of extracellular domain using monoclonal antibodies . Cetuximab is the most developed one and there is plenty information on the literature about its current status . In this review we focus on other P00533 monoclonal antibodies under clinical development . The more developed one is DB01269 . Its clinical development is taking place very quickly and it has mainly been studied in colorectal cancer showing promising results . There are also other interesting drugs such as DB05101 , Nimotuzumab and Zalutumumab . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . Cortical neurons from intrauterine growth retardation rats exhibit lower response to neurotrophin P23560 . Intrauterine growth retardation ( IUGR ) is putatively involved in the pathophysiology of schizophrenia . The animal model of IUGR induced by synthetic thromboxane A2 ( TXA2 ) is useful to clarify the effect of IUGR on pups ' brains , however , analysis at the cellular level is still needed . P23560 ( P23560 ) , which plays a role in neuronal survival and synaptic plasticity in the central nervous system ( CNS ) , may also be associated with schizophrenia . However , the possible relationship between IUGR and P23560 function remains unclear . Here , we examined how IUGR by TXA2 impacts P23560 function by using dissociated cortical neurons . We found that , although P23560 levels in cultured neurons from the cerebral cortex of low birth weight pups with IUGR were unchanged , TrkB ( P23560 receptor ) was decreased compared with control-rats . P23560 -stimulated MAPK/ P27361 /2 and PI3K/Akt pathways , which are downstream intracellular signaling pathways of TrkB , were repressed in IUGR-rat cultures . Expression of glutamate receptors such as P42261 and Q12879 was also suppressed in IUGR-rat cultures . Furthermore , in IUGR-rat cultures , anti-apoptotic protein Bcl2 was decreased and P23560 failed to prevent neurons from cell death caused by serum-deprivation . Taken together , IUGR resulted in reductions in cell viability and in synaptic function following TrkB down-regulation , which may play a role in schizophrenia-like behaviors . DB00193 and another atypical opioid meperidine have exaggerated serotonin syndrome behavioural effects , but decreased analgesic effects , in genetically deficient serotonin transporter ( P31645 ) mice . The serotonin syndrome is a potential side-effect of serotonin-enhancing drugs , including antidepressants such as selective serotonin reuptake inhibitors ( SSRIs ) and monoamine oxidase inhibitors ( MAOIs ) . We recently reported a genetic mouse model for the serotonin syndrome , as serotonin transporter ( P31645 ) -deficient mice have exaggerated serotonin syndrome behavioural responses to the MAOI tranylcypromine and the serotonin precursor 5-hydroxy-l-tryptophan ( 5-HTP ) . As numerous case reports implicate the atypical opioids tramadol and meperidine in the development of the human serotonin syndrome , we examined tramadol and meperidine as possible causative drugs in the rodent model of the serotonin syndrome in P31645 wild-type ( +/+ ) , heterozygous ( +/- ) and knockout ( -/- ) mice . Comparisons were made with P31645 mice treated with either vehicle or morphine , an opioid not implicated in the serotonin syndrome in humans . Here we show that tramadol and meperidine , but not morphine , induce serotonin syndrome-like behaviours in mice , and we show that this response is exaggerated in mice lacking one or two copies of P31645 . The exaggerated response to tramadol in P31645 -/- mice was blocked by pretreatment with the P08908 antagonist WAY 100635 . Further , we show that morphine- , meperidine- and tramadol-induced analgesia is markedly decreased in P31645 -/- mice . These studies suggest that caution seems warranted in prescribing or not warning patients receiving SSRIs or MAOIs that dangerous side-effects may occur during concurrent use of tramadol and similar agents . These findings suggest that it is conceivable that there might be increased vulnerability in individuals with P31645 polymorphisms that may reduce P31645 by more than 50 % , the level in P31645 +/- mice . Molecular biology of gallbladder cancer : potential clinical implications . Gallbladder cancer ( GBC ) is a common malignancy of the biliary tract and involves the changes in multiple oncogenes and multiple genetic genes . Since over the past decade there has been an advance in the knowledge of the genetic basis of cancer , mainly as a result of the rapid progression of molecular technology ; however , conventional therapeutic approaches have not had much impact on the course of this aggressive neoplasm . Knowledge of the molecular biology of GBC is rapidly growing . Genetic alterations in GBC include adenosine triphosphate-binding cassette transporter Q9H221 , membrane-bound enzyme ADAM-17 of multi-functional gene family , and other genes including p53 , P35354 , Q01831 , and RASSF1A . The advances in molecular biology have potential implications for the detection of this disease , using Synuclein-gamma , P18827 , glycoprotein 72 ( TAG-72 ) , tumor endothelial marker 8 protein ( Q9H6X2 ) and P01375 . The use of these molecular diagnostic methods is of clinical importance for the gene replacement therapy , genetic prodrug activation therapy , and antisense immunology technology for the treatment of malignancy . The author reviewed recent publications on PubMed , and summarized molecular biology of GBC , with an emphasis on features of potential clinical implications for diagnosis and management . [ Effects of chronic fluoxetine treatment on catalepsy and immune response in mice genetically predisposed to freezing reaction : the role of P08908 and 5- Q13049 receptors and tph2 and P31645 genes ] . ASC/Icg ( Antidepressant Sensitive Catalepsy ) mouse strain selected for high predisposition to pinch-induced catalepsy is characterized by depressive-like behavior and impaired immune response . Chronic treatment with SSRI fluoxetine attenuated catalepsy manifestation and normalized a decreased number of rosette-forming cells ( P41440 ) in spleen in ASC mice . Chronic fluoxetine administration had no effect on catalepsy and P41440 number in mice of parental cataleptic CBA/Lac strain . DB00472 failed to alter P08908 receptor functional activity in mice of both strains and diminished 5- Q13049 receptor functional activity in CBA but not in ASC mice . No effect on cortical P08908 and 5- Q13049 receptor mRNA levels and on P08908 receptor , tph2 ( tryptophan hydroxylase-2 ) and P31645 ( serotonin transporter ) mesencephalic gene expression was observed in ASC mice . Other possible serotonergic mechanisms of fluoxetine effect on catalepsy and immune response in mice with depressive-like state are discussed . P11511 inhibitors and cyclooxygenase-2 ( P35354 ) inhibitors in endometriosis : new questions -- old answers ? The medical treatment of endometriosis needs to be optimized . Therapeutic management strategies for endometriosis-associated pain or recurrent disease are primarily aimed at downregulating ovarian function or antagonizing the effect of estrogen in ectopic endometrial implants . In this context , basic research is providing important results for the development of new , specific treatment modalities . P11511 overexpression has recently been detected in endometriotic tissue . P11511 ( p450arom ) is responsible for converting C19 androgens into estrogen in several types of human tissue . P11511 activity causes local estrogen biosynthesis , which , in turn , stimulates prostaglandin E2 production by upregulating cyclooxygenase-2 ( P35354 ) . Thus , a positive feedback cycle develops between the two systems . Another abnormality in endometriosis , the deficient 17beta-hydroxysteroiddehydrogenase type II ( 17beta-HSD-Type-II ) expression , impairs the inactivation of estradiol to estrone . In contrast to the eutopic endometrium , these molecular aberrations increase the amount of local estradiol and prostaglandin E2 in endometriosis . In several human cell lines , prostaglandin and estrogen concentrations are associated with proliferation , migration , angiogenesis , apoptosis resistance and even invasiveness . Consequently , aromatase and P35354 are thought to be promising new therapeutic targets . Thus , specific aromatase inhibitors ( e.g. DB01006 / DB01006 , DB01217 /Arimidex or Exemestan/Aromasin ) or selective P35354 inhibitors ( e.g. Celecoxib/ DB00482 , DB00533 /Vioxx , DB00580 /Bextra ) are of great interest and should be studied in clinical trials in premenopausal woman with endometriosis to expand the spectrum of currently available treatment options . Inhibition of P04275 -mediated platelet activation and thrombosis by the anti- P04275 A1-domain aptamer DB05202 . BACKGROUND : P04275 ( P04275 ) has a role in both hemostasis and thrombosis . Platelets adhere to damaged arteries by interactions between the P04275 A1-domain and glycoprotein Ib receptors under conditions of high shear . This initial platelet binding event stimulates platelet activation , recruitment , and activation of the clotting cascade , promoting thrombus formation . OBJECTIVE : To characterize the inhibitory activity of a P04275 inhibitory aptamer . METHODS : Using in vitro selection , aptamer stabilization , and conjugation to a 20-kDa poly ( ethylene glycol ) , we generated a nuclease-resistant aptamer , DB05202 , that binds to the P04275 A1-domain with high affinity ( K(D) approximately 2 nM ) . The aptamer was assessed for inhibition of P04275 -induced platelet aggregation . In vitro inhibition of platelet adhesion was assessed on collagen-coated slides and injured pig aortic segments . In vivo activity was assessed in a cynomolgus monkey carotid electrical injury thrombosis model . RESULTS AND CONCLUSION : DB05202 inhibited botrocetin-induced platelet aggregation ( IC90 approximately 300 nM ) and shear force-induced platelet aggregation ( IC95 approximately 400 nM ) . It reduced adhesion of platelets to collagen-coated matrices and formation of platelet thrombi on denuded porcine arteries . DB05202 also inhibited the formation of occlusive thrombi in cynomolgus monkeys . We have discovered a novel anti- P04275 aptamer that could have therapeutic use as an anti- P04275 agent in the setting of P04275 -mediated thrombosis .

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MH_train_1360

interacts_with DB00422?

multiple_choice

Neuregulin and dopamine modulation of hippocampal gamma oscillations is dependent on dopamine D4 receptors . The neuregulin/ErbB signaling network is genetically associated with schizophrenia and modulates hippocampal γ oscillations -- a type of neuronal network activity important for higher brain processes and altered in psychiatric disorders . Because neuregulin-1 ( Q99988 ) dramatically increases extracellular dopamine levels in the hippocampus , we investigated the relationship between DB04223 /ErbB and dopamine signaling in hippocampal γ oscillations . Using agonists for different D1- and D2-type dopamine receptors , we found that the D4 receptor ( D4R ) agonist PD168077 , but not D1/D5 and D2/D3 agonists , increases γ oscillation power , and its effect is blocked by the highly specific D4R antagonist L-745,870 . Using double in situ hybridization and immunofluorescence histochemistry , we show that hippocampal D4R mRNA and protein are more highly expressed in Q99259 -positive GABAergic interneurons , many of which express the Q99988 receptor ErbB4 . Importantly , D4 and ErbB4 receptors are coexpressed in parvalbumin-positive basket cells that are critical for γ oscillations . Last , we report that D4R activation is essential for the effects of Q99988 on network activity because L-745,870 and the atypical antipsychotic clozapine dramatically reduce the Q99988 -induced increase in γ oscillation power . This unique link between D4R and ErbB4 signaling on γ oscillation power , and their coexpression in parvalbumin-expressing interneurons , suggests a cellular mechanism that may be compromised in different psychiatric disorders affecting cognitive control . These findings are important given the association of a P21917 polymorphism with alterations in attention , working memory , and γ oscillations , and suggest potential benefits of D4R modulators for targeting cognitive deficits . Identification and localization of DNA alteration in Chinese hamster ovary cell mutants ( Urd- ) defective in the first three enzymes of de novo pyrimidine synthesis . In animals , the first three enzymatic steps of de novo pyrimidine synthesis , carbamyl phosphate synthetase , aspartate transcarbamylase , and dihydroorotase , comprise the multifunctional protein known as the P27708 . Mutants of Chinese hamster ovary cells ( CHO- P04264 , pro- ) deficient in P27708 activities require uridine for growth and are designated Urd-A mutants . To examine further the nature of the genetic alterations in Urd-A mutants and revertants , we have performed a detailed Southern blot hybridization analysis of DNA from wild-type , Urd-A , and revertant cells using as hybridization probes cDNAs complementary to CAD mRNA isolated from Syrian hamster . This has allowed us to identify an apparent alteration in the CAD gene in DNA from Urd-A cells . This alteration is in a region of the gene which appears to correspond to the region of the protein which is hypersensitive to proteases and which seems to be altered in the mutants . Only one of the two CAD alleles present appears to be altered in this way . Study of certain revertants of Urd-A strongly suggests that in some cases reversion has occurred by amplification of the mutant CAD allele . Determination of glutamate decarboxylase by high-performance liquid chromatography . An improved method for the determination of glutamate decarboxylase ( Q99259 ) activity is described . The enzyme was evaluated by incubation with glutamic acid ( L- DB00142 ) in the presence of pyridoxal 5'-phosphate ( PLP ) : the gamma-aminobutyric acid ( GABA ) formed was derivatized to PTC-GABA ; the latter was subsequently separated and assayed by isocratic HPLC ( LiChrospher RP-18 column ; isocratic elution with pH 5.8 acetate buffer in acetonitrile-water ) with UV absorbance detection at 254 nm . The method described is a sensitive , reproducible and specific assay useful for following variations of Q99259 activity in vitro ; this assay was subsequently used for the evaluation of Q99259 activity variations after irradiation with low doses of He-Ne laser radiation . Accelerated and long-term hematopoietic engraftment in mice transplanted with ex vivo expanded bone marrow . Using a murine experimental model , we investigated whether ex vivo expansion of BM grafts under P08700 / P05231 stimulation accelerates the early hematopoietic recovery of recipients of BM transplants . To facilitate the ex vivo expansion of hematopoietic progenitors , BM was first enriched in proliferatively active hematopoietic stem cells by a single treatment with 5-fluorouracil ( 5FU ) 4 days prior to the BM harvest . The results showed that the number of CFU-GM and CFU- P28222 progenitors in the graft was significantly increased ( 56-fold and 14-fold , respectively ) , as a result of a 3 day incubation in the presence of P08700 and P05231 . Daily analysis of animals transplanted with 5 x 10(4) BM cells , either freshly harvested or expanded for 3 days , showed that the expanded grafts consistently allowed a faster hematopoietic recovery of recipients . Differences between both groups of transplanted animals were most evident when the number of either femoral or splenic CFU-GMs were compared , with increases close to 70-fold at the fifth day of engraftment being observed . Similarly , mice transplanted with expanded grafts showed a hastened recovery in the cellularity of both organs that was most significant during the second week following transplantation , with maximal increases of 15 and 40-fold in the BM and spleen , respectively . Differences in peripheral leukocyte numbers between both groups of recipients were much less remarkable than those observed in the hematopoietic organs , although from the nadir period to the 11th day post-transplantation differences ranging from twofold to sixfold were apparent , consistent with a higher rate of mouse survival. ( ABSTRACT TRUNCATED AT 250 WORDS ) MnSOD drives neuroendocrine differentiation , androgen independence , and cell survival in prostate cancer cells . An increase in neuroendocrine ( NE ) cell number has been associated with progression of prostate tumor , one of the most frequent cancers among Western males . We previously reported that mitochondrial manganese superoxide dismutase ( MnSOD ) increases during the NE differentiation process . The goal of this study was to find whether MnSOD up-regulation is enough to induce NE differentiation . Several human prostate cancer LNCaP cell clones stably overexpressing MnSOD were characterized and two were selected ( MnSOD-S4 and MnSOD- P28222 ) . MnSOD overexpression induces NE morphological features as well as coexpression of the NE marker synaptophysin . Both MnSOD clones exhibit lower superoxide levels and higher H(2)O(2) levels . MnSOD-overexpressing cells show higher proliferation rates in complete medium , but in steroid-free medium MnSOD- P28222 cells are still capable of proliferation . MnSOD up-regulation decreases androgen receptor and prevents its nuclear translocation . MnSOD also induces up-regulation of Bcl-2 and prevents docetaxel- , etoposide- , or P01375 -induced cell death . Finally , MnSOD-overexpressing cells enhance growth of androgen-independent PC-3 cells but reduce growth of androgen-dependent cells . These results indicate that redox modulation caused by MnSOD overexpression explains most NE-like features , including morphological changes , NE marker expression , androgen independence , inhibition of apoptosis , and enhancement of cell growth . Many of these events can be associated with the androgen dependent-independent transition during prostate cancer progression . Development of peptidomimetic ligands of Pro- DB00149 - DB00145 -NH(2) as allosteric modulators of the dopamine D(2) receptor . A variety of stable , small-molecule peptidomimetic ligands have been developed to elucidate the mechanism by which the neuropeptide Pro- DB00149 - DB00145 -NH(2) ( P00747 ) modulates dopaminergic neurotransmission . Photoaffinity labeling ligands based upon P00747 peptidomimetics have been used to establish that P00747 binds to the P14416 at a site that is different from the orthosteric site , thus making P00747 and its peptidomimetics allosteric modulators of the dopamine receptor . Through the design , synthesis and pharmacological evaluation of conformationally constrained peptidomimetics containing lactam , bicyclic , and spiro-bicyclic scaffolds , support was provided for the hypothesis that the bioactive conformation of P00747 is a type II β-turn . In addition , studies with peptidomimetics designed to mimic either a type VI β-turn or polyproline II helix conformation yielded molecules that were able to modulate dopamine receptors because of their ability to place the carboxamide NH(2) pharmacophore in the same topological space as that seen in the type II β-turn . Extensive studies with the spiro-bicyclic P00747 peptidomimetics also established that both positive and negative modes of modulation were possible for the same series of peptidomimetics simply as a result of minor differences in the stereochemistry about the bridgehead carbon within the scaffold . This information was used to transform existing positive modulators into negative modulators , which demonstrated that small structural changes in the spiro-bicyclic dopamine receptor modulators are capable of causing major changes in the modulatory activity of P00747 peptidomimetics . Prion protein stimulates tissue-type plasminogen activator-mediated plasmin generation via a lysine-binding site on kringle 2 . Recombinant human prion-protein ( PrP23-231 ) stimulates plasminogen activation by tissue-type plasminogen activator ( t-PA ) . The stimulatory activity is conserved in the N-terminal fragment ( PrP23-110 ) . It has further been shown by others that P04156 (c) binds to kringle-domains of plasminogen . We compared the stimulatory activity of recombinant PrP23-231 and PrP23-110 on plasminogen activation catalyzed by t-PA , urokinase ( u-PA ) , streptokinase and Desmodus salivary plasminogen activator ( DSPAalpha1 ) . As these plasminogen activators are distinct , with respect to their kringle domains we studied their binding to immobilized PrP23-110 . P00747 activation was measured in a chromogenic assay in vitro and binding studies were carried out using surface plasmon resonance technology . We found that recombinant full-length prion protein , PrP23-231 , and PrP23-110 specifically stimulate t-PA mediated plasminogen activation . Two hundred nanomoles per liter of PrP23-110 stimulated 1.8 nmol L(-1) t-PA 48-fold , 180 nmol L(-1) DB04925 (alpha1) 2.5-fold , 1.8 nmol L(-1) u-PA 1.1-fold , and 1.8 nmol L(-1) streptokinase 1.8-fold . Our data show no specific binding for streptokinase . In contrast all plasminogen activators carrying a kringle domain bound to PrP23-110 . We further studied the effect of lysine on binding to PrP23-110 and on plasminogen activation by DB04925 (alpha1) or t-PA . DB00123 decreased both the binding of t-PA to PrP23-110 and the stimulation of plasmin generation by t-PA . Both binding and plasminogen activation of DB04925 (alpha1) were not influenced by the presence of lysine . All plasminogen activators tested bearing kringle domains bind to PrP23-110 . Binding to PrP23-110 is not sufficient for stimulation of plasmin generation . Thus the lysine-binding site of kringle 2 that is unique to t-PA appears to mediate the specific stimulation of plasminogen activation by the cellular prion protein . Positive effects of methylphenidate on hyperactivity are moderated by monoaminergic gene variants in children with autism spectrum disorders . Methylphenidate ( DB00422 ) reduces hyperactive-impulsive symptoms common in children with autism spectrum disorders ( ASDs ) , however , response and tolerability varies widely . We hypothesized monoaminergic gene variants may moderate DB00422 effects in P51689 , as in typically developing children with attention-deficit/hyperactivity disorder . Genotype data were available for 64 children with P51689 and hyperactivity who were exposed to DB00422 during a 1-week safety/tolerability lead-in phase and 58 who went on to be randomized to placebo and three doses of DB00422 during a 4-week blinded , crossover study . Outcome measures included the Clinical Global Impression-Improvement ( CGI-I ) scale and the Aberrant Behavior Checklist ( ABC-hyperactivity index ) . A total of 14 subjects discontinued the study because of DB00422 side effects . Subjects were genotyped for variants in P21728 - P21918 , P08913 , Q01959 , P31645 , P21397 and P27338 , and P21964 . Forty-nine percent of the sample met positive responder criteria . In this modest but relatively homogeneous sample , significant differences by P21728 ( P=0.006 ) , P08913 ( P < 0.02 ) , P21964 ( P < 0.04 ) , P35462 ( P < 0.05 ) , P21917 ( P < 0.05 ) , Q01959 ( P < 0.05 ) and P31645 ( P < 0.05 ) genotypes were found for responders versus non-responders . Variants in P14416 ( P < 0.001 ) and P35462 ( P < 0.04 ) were associated with tolerability in the 14 subjects who discontinued the trial . For this first DB00422 pharmacogenetic study in children with P51689 , multiple monoaminergic gene variants may help explain individual differences in DB00422 's efficacy and tolerability . Identification of an amino acid residue important for binding of methiothepin and sumatriptan to the human 5-HT(1B) receptor . Site-directed mutagenesis of the human P28222 receptor was performed to investigate the role of the amino acid residues cysteine 326 and tryptophan 327 in transmembrane region VI and aspartic acid 352 in transmembrane region VII in ligand binding . Binding studies were performed with the antagonist radioligand [3H]GR125743 on mutant and wild-type receptors stably expressed in Chinese hamster ovary cells ( CHO ) - P04264 cells . Substitution of tryptophan 327 by alanine resulted in decreased affinities of all ligands tested . The most prominent changes in affinity were observed for the antagonist methiothepin and the antimigraine drug sumatriptan , which were reduced approximately 300- and 60-fold , respectively . Nevertheless , the affinity of 5-HT remained the same . Replacement of the aspartic acid 352 by alanine reduced high-affinity binding of 5-HT . Substitution of cysteine 326 by alanine had minor effects on ligand binding . Some of these results agree with the results from mutagenesis studies of the corresponding amino acids in other receptors . However , some notable differences also emerge showing that functional roles of individual amino acid residues must be tested experimentally in each receptor subtype . Serial changes in serum vitamin P04264 , triglyceride , cholesterol , osteocalcin and 25-hydroxyvitamin D3 in patients after hip replacement for fractured neck of femur or osteoarthritis . Serum vitamin P04264 concentrations were measured at presentation ( just before surgery ) and then at weekly intervals for 3 weeks in two groups of elderly patients requiring either hemiarthroplasty for fractured neck of femur ( FON , n = 13 ) or total hip replacement for osteoarthritis of the hip ( OA , n = 16 ) . In comparison with healthy elderly volunteers ( n = 25 ) , serum vitamin P04264 concentrations were significantly lower in both groups at presentation , and fell significantly within 24 h after surgery to concentrations approaching non-detectable , subsequently returning to pre-operative values within 3 weeks . Serum vitamin P04264 tended to be lower in the fracture group both before and after operation , although calculation of a vitamin P04264 -triglyceride ratio reduced the apparent difference as triglyceride concentrations were lower in the fracture group . P02818 concentrations were similar and fell significantly after operation in both groups , returning to pre-operative levels within 7 days . No differences in the two forms of osteocalcin ( carboxylated and undercarboxylated ) were observed either before or after operation in either group . DB00146 concentrations were not significantly different between the two groups at any time . DB01022 status may be lower than desirable in certain groups of the elderly population , and supplementation should be considered as prophylactic therapy . No significant association between genetic variants in 7 candidate genes and response to methylphenidate treatment in adult patients with ADHD . Results from pharmacogenetic investigations of methylphenidate ( DB00422 ) response in patients with ADHD are still inconsistent , especially among adults . This study investigates the role of genetic variants ( P31645 , P28222 , Q8IWU9 , P09172 , P21917 , P21964 , and P60880 ) in the response to DB00422 in a sample of 164 adults . Genes were chosen owing to previous evidence for an influence in ADHD susceptibility . No significant differences in allele or genotype frequencies between DB00422 responders and nonresponders were detected . In conclusion , our findings do not support an effect of these genes in the pharmacogenetics of DB00422 among adults with ADHD . DB05651 , a novel isotype-selective histone deacetylase inhibitor , has broad spectrum antitumor activity in vitro and in vivo . Nonselective inhibitors of human histone deacetylases ( HDAC ) are known to have antitumor activity in mice in vivo , and several of them are under clinical investigation . The first of these , DB02546 ( DB02546 ) , has been approved for treatment of cutaneous T-cell lymphoma . Questions remain concerning which HDAC isotype(s) are the best to target for anticancer activity and whether increased efficacy and safety will result with an isotype-selective HDAC inhibitor . We have developed an isotype-selective HDAC inhibitor , DB05651 , which potently targets human Q13547 but also has inhibitory activity against Q92769 , O15379 , and Q96DB2 in vitro . In intact cells , DB05651 inhibited only a fraction of the total HDAC activity and showed long-lasting inhibitory activity even upon drug removal . DB05651 induced hyperacetylation of histones , selectively induced apoptosis , and caused cell cycle blockade in various human cancer cell lines in a dose-dependent manner . DB05651 exhibited potent and selective antiproliferative activities against a broad spectrum of human cancer cell lines in vitro , and HDAC inhibitory activity was required for these effects . In vivo , DB05651 significantly inhibited growth of human tumor xenografts in nude mice in a dose-dependent manner and the antitumor activity correlated with induction of histone acetylation in tumors . Our findings suggest that the isotype-selective HDAC inhibition by DB05651 is sufficient for antitumor activity in vivo and that further clinical investigation is warranted . Inhibition of the renin-angiotensin system improves physiological outcomes in mice with mild or severe cancer cachexia . Cancer cachexia describes the progressive skeletal muscle wasting and weakness associated with many cancers . Cachexia reduces mobility and quality of life and accounts for 20-30 % of all cancer-related deaths . Activation of the renin-angiotensin system causes skeletal muscle wasting and weakness . We tested the hypothesis that treatment with the angiotensin converting enzyme ( P12821 ) inhibitor , perindopril , would enhance whole body and skeletal muscle function in cachectic mice bearing Colon-26 ( C-26 ) tumors . CD2F1 mice received a subcutaneous injection of phosphate buffered saline or C-26 tumor cells inducing either a mild or severe cachexia . The following day , one cohort of C-26 mice began receiving perindopril in their drinking water ( 4 mg kg(-1) day(-1) ) for 21 days . In mild and severe cachexia , perindopril increased measures of whole body function ( grip strength and rotarod ) and reduced fatigue in isolated contracting diaphragm muscle strips ( p < 0.05 ) . In severely cachectic mice , perindopril reduced tumor growth , improved locomotor activity and reduced fatigue of tibialis anterior muscles in situ ( p < 0.05 ) , which was associated with increased oxidative enzyme capacity ( succinate deyhydrogenase , p < 0.05 ) . DB00790 attenuated the increase in Q969Q1 and P05231 mRNA expression and enhanced Akt phosphorylation in severely cachectic mice but neither body nor muscle mass was increased . These findings support the therapeutic potential of P12821 inhibition for enhancing whole body function and reducing fatigue of respiratory muscles in early and late stage cancer cachexia and should be confirmed in future clinical trials . Since P12821 inhibition alone did not enhance body or muscle mass , co-treatment with an anabolic agent may be required to address these aspects of cancer cachexia . Effects of histone deacetylase inhibitors on amygdaloid histone acetylation and neuropeptide Y expression : a role in anxiety-like and alcohol-drinking behaviours . Recent studies have demonstrated the involvement of epigenetic mechanisms in psychiatric disorders , including alcoholism . Here , we investigated the effects of histone deacetylase ( HDAC ) inhibitor , trichostatin A ( P32119 ) on amygdaloid HDAC-induced histone deacetylation and neuropeptide Y ( P01303 ) expression and on anxiety-like and alcohol-drinking behaviours in alcohol-preferring ( P ) and -non-preferring ( NP ) rats . It was found that P rats displayed higher anxiety-like and alcohol-drinking behaviours , higher amygdaloid nuclear , but not cytosolic , HDAC activity , which was associated with increased Q92769 protein levels and deficits in histone acetylation and P01303 expression in the central ( CeA ) and medial nucleus of amygdala ( MeA ) , as compared to NP rats . P32119 treatment attenuated the anxiety-like and alcohol-drinking behaviours , with concomitant reductions in amygdaloid nuclear , but not cytosolic HDAC activity , and Q92769 , but not P56524 , protein levels in the CeA and MeA of P rats , without effect in NP rats . P32119 treatment also increased global histone acetylation ( H3- P35527 and H4-K8 ) and P01303 expression in the CeA and MeA of P , but not in NP rats . Histone H3 acetylation within the P01303 promoter was also innately lower in the amygdala of P rats compared with NP rats ; which was normalized by P32119 treatment . Voluntary ethanol intake in P , but not NP rats , produced anxiolytic effects and decreased the Q92769 levels and increased histone acetylation in the CeA and MeA . These results suggest that higher Q92769 expression-related deficits in histone acetylation may be involved in lower P01303 expression in the amygdala of P rats , and operative in controlling anxiety-like and alcohol-drinking behaviours . Characterization of DB05005 -B , an orally bioavailable antagonist of the P51686 chemokine receptor , for treatment of inflammatory bowel disease . The chemokine system represents a diverse group of G protein-coupled receptors responsible for orchestrating cell recruitment under both homeostatic and inflammatory conditions . Chemokine receptor 9 ( P51686 ) is a chemokine receptor known to be central for migration of immune cells into the intestine . Its only ligand , O15444 , is expressed at the mucosal surface of the intestine and is known to be elevated in intestinal inflammation . To date , there are no reports of small-molecule antagonists targeting P51686 . We report , for the first time , the discovery of a small molecule , DB05005 -B , which is an orally bioavailable , selective , and potent antagonist of human P51686 . DB05005 -B inhibited P51686 -mediated Ca(2+) mobilization and chemotaxis on Molt-4 cells with IC(50) values of 5.4 and 3.4 nM , respectively . In the presence of 100 % human serum , DB05005 -B inhibited P51686 -mediated chemotaxis with an IC(50) of 33 nM , and the addition of α1-acid glycoprotein did not affect its potency . DB05005 -B inhibited chemotaxis of primary P51686 -expressing cells to O15444 with an IC(50) of 6.8 nM . DB05005 -B was an equipotent inhibitor of O15444 -directed chemotaxis of both splice forms of P51686 ( CCR9A and CCR9B ) with IC(50) values of 2.8 and 2.6 nM , respectively . DB05005 -B also inhibited mouse and rat P51686 -mediated chemotaxis . Inhibition of P51686 with DB05005 -B results in normalization of Crohn 's disease such as histopathology associated with the P01375 (ΔARE) mice . Analysis of the plasma level of drug associated with this improvement provides an understanding of the pharmacokinetic/pharmacodynamic relationship for P51686 antagonists in the treatment of intestinal inflammation . Discovery and validation of methylation markers for endometrial cancer . The prognosis of endometrial cancer is strongly associated with stage at diagnosis , suggesting that early detection may reduce mortality . Women who are diagnosed with endometrial carcinoma often have a lengthy history of vaginal bleeding , which offers an opportunity for early diagnosis and curative treatment . We performed DNA methylation profiling on population-based endometrial cancers to identify early detection biomarkers and replicated top candidates in two independent studies . We compared DNA methylation values of 1,500 probes representing 807 genes in 148 population-based endometrial carcinoma samples and 23 benign endometrial tissues . Markers were replicated in another set of 69 carcinomas and 40 benign tissues profiled on the same platform . Further replication was conducted in The Cancer Genome Atlas and in prospectively collected endometrial brushings from women with and without endometrial carcinomas . We identified 114 CpG sites showing methylation differences with p values of ≤ 10(-7) between endometrial carcinoma and normal endometrium . Eight genes ( P18509 , Q99929 , Q9Y278 , P28222 , MME , P01303 and O00570 ) were selected for further replication . Age-adjusted odds ratios for endometrial cancer ranged from 3.44 ( 95 % -CI : 1.33-8.91 ) for Q99929 to 18.61 ( 95 % -CI : 5.50-62.97 ) for P28222 . An area under the curve ( AUC ) of 0.93 was achieved for discriminating carcinoma from benign endometrium . Replication in The Cancer Genome Atlas and in endometrial brushings from an independent study confirmed the candidate markers . This study demonstrates that methylation markers may be used to evaluate women with abnormal vaginal bleeding to distinguish women with endometrial carcinoma from the majority of women without malignancy . DB02424 attenuates 3‑nitropropionic acid‑induced apoptosis and JNK activation through the expression of HSP 70 in striatal cells . Although selective striatal cell death is a characteristic hallmark in the pathogenesis of Huntington 's disease ( HD ) , the underlying mechanism of striatal susceptibility remains to be clarified . Heat shock proteins ( HSPs ) have been reported to suppress the aggregate formation of mutant huntingtin and concurrent striatal cell death . In a previous study , we observed that heat shock transcription factor 1 ( Q00613 ) , a major transcription factor of HSPs , significantly attenuated 3‑nitropropionic acid (3NP)‑induced reactive oxygen species ( ROS ) production and apoptosis through the expression of HSP 70 in striatal cells . To investigate the differential roles of HSPs in 3NP‑induced striatal cell death , the effect of geldanamycin ( GA ) , an P08238 inhibitor , was examined in 3NP‑stimulated striatal cells . GA significantly attenuated 3NP‑induced striatal apoptosis and ROS production with an increased expression of HSP 70 . Triptolide ( TL ) , an HSP 70 inhibitor , abolished GA‑mediated protective effects in 3NP‑stimulated striatal cells . To understand the underlying mechanism by which GA‑mediated HSP 70 protects striatal cells against 3NP stimulation , the involvement of various signaling pathways was examined . GA significantly attenuated 3NP‑induced c‑Jun N‑terminal kinase ( JNK ) phosphorylation and subsequent c‑Jun phosphorylation in striatal cells . Taken together , the present study demonstrated that GA exhibits protective properties against 3NP‑induced apoptosis and JNK activation via the induction of HSP 70 in striatal cells , suggesting that expression of HSP 70 may be a valuable therapeutic target in the treatment of HD . The cloning and reintroduction into animal cells of a functional CAD gene , a dominant amplifiable genetic marker . Rodent cells resistant to DB03459 , a specific inhibitor of the aspartate transcarbamylase activity of the multifunctional P27708 , overproduce CAD as a result of amplification of the CAD gene . We cloned a functional CAD gene from Syrian hamster cells using a cosmid vector . Two independently isolated cosmids containing CAD genes have inserts 40 and 45 kb long . We introduced the cloned genes into CAD-deficient Chinese hamster ovary ( CHO ) cell mutants by fusing them with protoplasts of Escherichia coli containing the cosmids . We also introduced the cloned genes into wild-type CHO cells by selecting cells that became resistant to high concentrations of DB03459 following protoplast fusion . The transformants of the mutant and wild-type CHO cells contain multiple active copies of the donated Syrian hamster CAD genes . The cloned genes in three independent transformants are integrated into host-cell chromosomes at single locations identified by in situ hybridization . In two of these transformants , the genes are located in one X chromosome or in a chromosome resembling the X . In the third case , the genes are located in a small metacentric or rearranged chromosome . Cloning , expression , and characterization of chicken transforming growth factor beta 4 . Transforming growth factor beta 4 ( TGF-beta 4 ) is unique to avian species , though its roles in vivo have not yet been well established . In this paper we describe the expression and partial characterization of recombinant chicken TGF-beta 4 . By using a GC-rich PCR system in a modified 5'RACE methodology we generated the 5'-end of cDNA sequence encoding the TGF-beta 4 precursor , which was in-frame cloned into pcDNA3.1/V5- DB00117 -TOPO and transfected into the Chinese hamster ovary cell line ( CHO- P04264 ) . A cell line stably expressing TGF-beta 4 precursor protein was established from CHO- P04264 cells . Acid-activated mature TGF-beta 4 inhibited the growth of mink lung epithelial ( Mv1Lu ) cell line . TGF-beta 4 also stimulated the expression of type I procollagen and enhanced heat shock protein 47 ( Hsp47 ) expression in chicken tendon fibroblasts . Hsp47 expression by TGF beta 4 is likely regulated through activation of heat shock transcription factor 1 ( Q00613 ) . Because the presence of TGF-beta 1 has not been documented in avian cells and our data show that TGF-beta 4 elicits biological activities in chicken tendon cells , which closely parallel that of TGF-beta 1 , we propose that TGF-beta 4 plays roles in avian species similar to what TGF-beta 1 plays in mammalian species . Stimulation of luteinizing hormone secretion by N-methyl-D,L-aspartic acid in the adult male guinea-pig : incomplete blockade by gonadotropin-releasing hormone receptor antagonism . Stimulation of luteinizing hormone ( LH ) secretion by N-methyl-D,L-aspartic acid ( Q13145 ) , reported for several mammalian species , is generally accepted to be mediated through stimulation of hypothalamic gonadotropin-releasing hormone ( DB00644 ) release . In view of a previously reported unexpected inhibitory action of Q13145 on DB00644 release from hypothalamic explant of intact male guinea-pigs , the aim of the present study was to assess the in vivo effects of Q13145 in the adult male guinea-pig . In the gonadally intact male , Q13145 ( 5 mg/animal ) elicited a robust LH secretion , which was blocked by the N-methyl-D-asparte-receptor antagonist DL-2-amino-5-phosphonovaleric acid ( AP-5,12 mg/animal ) . In the castrated male , Q13145 elicited only a marginal and inconsistent LH secretion . DB00050 ( DB00456 ) , a P30968 antagonist , administered intracardiacally 1 min or 45 min preceding bolus injection of Q13145 significantly reduced the LH response to Q13145 in the intact male . Surprisingly , following P30968 blockade with DB00456 , there still was a substantial residual serum LH response to Q13145 , while DB00456 completely abolished the serum LH response to high dose ( 1 microg or 10 microg ) guinea-pig DB00644 ( gpGnRH ) . These results indicate that Q13145 stimulates LH secretion in the gonadally intact male guinea-pig in vivo and that this effect is mediated in part through gpGnRH-independent mechanisms . Increased inducible nitric oxide synthase in lung carcinoma of smokers . BACKGROUND : Cigarette smoking is well known to play an important role in the development of lung cancer . Inducible nitric oxide synthase ( P35228 ) can either promote or inhibit cell proliferation and growth , which makes its role in the development of malignant tumors controversial . The relation between cigarette smoking and P35228 in human lung cancer is unknown . METHODS : The study examined the levels of P35228 /NO in nonsmall-cell lung cancer ( NSCLC ) tissues of smokers and nonsmokers and in NSCLC cells ( NCI-H23 ) treated by 4-(N-Methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone ( NNK ) , a potent tobacco-specific carcinogen . RESULTS : The level of P35228 /NO was significantly higher in lung cancer tissues of smokers than that of nonsmokers . Unlike P35228 /NO , the activity of caspase-3 was reduced in the former compared with the latter . The expression of the cleaved caspase-3 was deceased in NCI-H23 cells treated with S-Nitroso-N-acetylpenicillamine ( P60880 ) , an NO donor , whereas treatment with NG-methyl-L-arginine ( Q13145 ) , an NO inhibitor , caused an increase in cleaved caspase-3 . Consistent with the change in caspase-3 , P60880 treatment inhibited cell death induced by UCN01 , a potent cell death-inducer . Q13145 treatment greatly enhanced the sensitivity of the cells to UCN01 . Further , the cells treated by NNK showed an increase in P35228 protein , accompanied by an elevation of cell proliferation . CONCLUSIONS : The study demonstrates that cigarette smoking promotes the level of P35228 /NO but suppresses the activity of caspase-3 , which may lead to the proliferation and growth of lung cancer cells .

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interacts_with DB01259?

multiple_choice

Reverse-phase protein array for prediction of patients at low risk of developing bone metastasis from breast cancer . BACKGROUND : A biomarker that predicts bone metastasis based on a protein laboratory assay has not been demonstrated . Reverse-phase protein array ( RPPA ) enables quantification of total and phosphorylated proteins , providing information about their functional status . The aim of this study was to identify bone-metastasis-related markers in patients with primary breast cancer using RPPA analysis . PATIENTS AND METHODS : Tumor samples were obtained from 169 patients with primary invasive breast carcinoma who underwent surgery . The patients were categorized by whether they developed breast cancer bone metastasis ( BCBM ) during follow-up . Clinical characteristics and protein expression by RPPA were compared and verified by leave-one-out cross-validation . RESULTS : Lymph node status ( p = .023 ) and expression level of 22 proteins by RPPA were significantly correlated with BCBM in logistic regression analysis . These variables were used to build a logistic regression model . After filtering the variables through a stepwise algorithm , the final model , consisting of 8 proteins and lymph node status , had sensitivity of 30.0 % , specificity of 90.5 % , positive predictive value of 30.0 % , and negative predictive value of 90.5 % in the cross-validation . Most of the identified proteins were associated with cell cycle or signal transduction ( P24941 , P38936 , Rb1 , Src , phosphorylated-ribosomal S6 kinase , P04626 , Q9H165 , and P35749 ) . CONCLUSION : Our validated model , in which the primary tumor is tested with RPPA , can predict patients who are at low risk of developing BCBM and thus who likely would not benefit from receiving a bisphosphonate in the adjuvant setting . Clinical trials excluding these patients have the potential to clarify the benefit of bisphosphonates in the adjuvant setting . Delta(9)- DB00470 enhances MCF-7 cell proliferation via cannabinoid receptor-independent signaling . We recently reported that Delta(9)-tetrahydrocannabinol ( Delta(9)-THC ) has the ability to stimulate the proliferation of human breast carcinoma MCF-7 cells . However , the mechanism of action remains to be clarified . The present study focused on the relationship between receptor expression and the effects of Delta(9)-THC on cell proliferation . RT-PCR analysis demonstrated that there was no detectable expression of CB receptors in MCF-7 cells . In accordance with this , no effects of cannabinoid 1/2 ( P21554 /2 ) receptor antagonists and pertussis toxin on cell proliferation were observed . Although MCF-7 cell proliferation is suggested to be suppressed by Delta(9)-THC in the presence of CB receptors , it was revealed that Delta(9)-THC could exert upregulation of living cells in the absence of the receptors . Interestingly , Delta(9)-THC upregulated human epithelial growth factor receptor type 2 ( P04626 ) expression , which is known to be a predictive factor of human breast cancer and is able to stimulate cancer cells as well as MCF-7 cells . DB00970 -treatment interfered with the upregulation of P04626 and cell proliferation by cannabinoid . Taken together , these studies suggest that , in the absence of CB receptors , Delta(9)-THC can stimulate the proliferation of MCF-7 cells by modulating , at least in part , P04626 transcription . Targeted treatment of advanced and metastaticbreast cancer with lapatinib . Improved molecular understanding of breast cancer in recent years has led to the discovery of important drug targets such as HER-2 and P00533 . DB01259 is a potent dual inhibitor of HER-2 and P00533 . Preclinical and phase I studies have shown activity with lapatinib in a number of cancers , including breast cancer , and the drug is well tolerated . The main known drug interactions are with paclitaxel and irinotecan . The most significant side-effects of lapatinib are diarrhea and adverse skin events . Rates of cardiotoxicity compare favorably with trastuzumab , a monoclonal antibody against HER-2 . This paper focuses on lapatinib in advanced and metastatic breast cancer , which remains an important therapeutic challenge . Phase II and III studies show activity as monotherapy , and in combination with chemotherapy or hormonal agents . Results from these studies suggest that the main benefit from lapatinib is in the HER-2 positive breast cancer population . Combinations of lapatinib and trastuzumab are also being studied and show encouraging results , particularly in trastuzumab-refractory metastatic breast cancer . DB01259 may have a specific role in treating HER-2 positive CNS metastases . The role of lapatinib as neoadjuvant therapy and in early breast cancer is also being evaluated . The novel DB01221 receptor antagonist , 2-hydroxy-5-(2,3,5,6-tetrafluoro-4-trifluoromethyl-benzylamino)-benzoic acid , is a gating modifier in cultured mouse cortical neurons . Neu2000 [ P04626 , 2-hydroxy-5-(2,3,5,6-tetrafluoro-4-trifluoromethyl-benzylamino)-benzoic acid ] , a derivative of sulfasalazine , attenuates DB01221 -induced neuronal toxicity . Here we investigated the effects of P04626 on the DB01221 receptor ( NMDAR ) using whole-cell patch clamp technique to determine the molecular mechanisms underlying its neuroprotective role . P04626 reversibly suppressed DB01221 responses in an uncompetitive manner with fast binding kinetics . Its inhibition of NMDAR activity depended on both the concentration and the use of agonist but not on the membrane potential . P04626 accelerated DB01221 desensitization without affecting the binding affinity of NMDAR for its agonists and stabilized the closed state of NMDAR . Therefore , P04626 should effectively alleviate disorders that are a result of glutamate excitoxicity with fewer side effects because it is a low-affinity gating modifier that antagonizes NMDAR in an uncompetitive manner . Moreover , in the presence of ifenprodil ( an Q13224 antagonist ) but not DB00238 -AAM077 [ ( R ) - [ ( S ) -1-(4-bromo-phenyl)-ethylamino ] -(2,3-dioxo-1,2,3,4-tetrahydro-quinoxalin-5-yl)-methyl ] -phosphonic acid , an Q12879 antagonist ] , the extent of P04626 block was decreased , suggesting that P04626 is an Q13224 -specific antagonist . SR147778 , a P21554 cannabinoid receptor antagonist , suppresses ethanol preference in chronically alcoholized Wistar rats . This study investigated the effect of the new P21554 cannabinoid receptor antagonist , SR147778 , on ethanol preference in chronically alcoholized Wistar rats . In study 1 , SR147778 , at doses of 0.3 , 1 , or 10 mg/kg/day ( mg/kg/d ) intraperitonealy ( ip ) , was administered during chronic pulmonary ethanol intoxication for 30 days . The rats were then exposed to a two-bottle choice ( ethanol 10 % v/v vs. water ) for at least 30 days . Neither 0.3 nor 1 mg/kg/d had any effect on ethanol preference . In contrast , the high dose induced a significant transient increase in ethanol intake between days 6 and 10 . In study 2 , SR147778 , at doses of 0.3 , 1 , or 10 mg/kg/d ip , was administered during the free-choice period after chronic alcoholization . Both ethanol preference and intake were significantly reduced only for 1 and 10 mg/kg/d . These results reinforce the hypothesis that the cannabinoid P21554 receptor is part of the neural substrate mediating alcohol intake and the motivational properties of alcohol . When these results are compared with those obtained with SR141716 ( DB06155 ) on ethanol preference , we observed that ( 1 ) coadministration of 10 mg/kg/d SR147778 during chronic alcoholization induced a shorter transient increase of ethanol intake than DB06155 and ( 2 ) SR147778 treatment during the free-choice period at doses of 1 and 10 mg/kg/d decreased ethanol intake more dramatically than SR141716 which , furthermore , continued for the duration of the free choice . Clinical development of eniluracil : current status . DB03516 is a potent inactivator of dihydropyrimidine dehydrogenase ( Q12882 ) , which is the first enzyme in the degradative pathway of systemically administered 5-fluorouracil ( DB00544 ) . Two completely oral regimens of eniluracil plus DB00544 are being evaluated in clinical trials : ( 1 ) a chronic schedule with both agents administered P55957 in a 10:1 ratio for 28 days of a 5-week course , and ( 2 ) a 5-day schedule of eniluracil once daily on days 1 through 7 and DB00544 once daily on days 2 through 6 . The clinical development of eniluracil is being pursued in several tumor types , including colorectal cancer , breast cancer , and pancreatic cancer . Response rates achieved in a phase II study of the chronic schedule of oral eniluracil/ DB00544 in patients with colorectal cancer compare favorably with those obtained in trials of intravenous DB00544 and leucovorin , while results from other trials are awaited . Safety analysis for the 28-day schedule has revealed a low incidence of severe toxicities , particularly as compared with standard DB00544 regimens . An LXR agonist promotes glioblastoma cell death through inhibition of an P00533 /AKT/ P36956 / P01130 -dependent pathway . Glioblastoma ( GBM ) is the most common malignant primary brain tumor of adults and one of the most lethal of all cancers . P00533 ( P00533 ) mutations ( EGFRvIII ) and phosphoinositide 3-kinase ( PI3K ) hyperactivation are common in GBM , promoting tumor growth and survival , including through sterol regulatory element-binding protein 1 ( P36956 ) -dependent lipogenesis . The role of cholesterol metabolism in GBM pathogenesis , its association with P00533 /PI3K signaling , and its potential therapeutic targetability are unknown . In our investigation , studies of GBM cell lines , xenograft models , and GBM clinical samples , including those from patients treated with the P00533 tyrosine kinase inhibitor lapatinib , uncovered an EGFRvIII-activated , PI3K/ P36956 -dependent tumor survival pathway through the low-density lipoprotein receptor ( P01130 ) . Targeting P01130 with the liver X receptor ( LXR ) agonist GW3965 caused inducible degrader of P01130 ( Q8WY64 ) -mediated P01130 degradation and increased expression of the O95477 cholesterol efflux transporter , potently promoting tumor cell death in an in vivo GBM model . These results show that EGFRvIII can promote tumor survival through PI3K/ P36956 -dependent upregulation of P01130 and suggest a role for LXR agonists in the treatment of GBM patients . Regional expression of prostaglandin E2 and F2alpha receptors in human myometrium , amnion , and choriodecidua with advancing gestation and labor . The change from uterine quiescence to enhanced contractile activity may be due to the differential expression of prostaglandin receptors within the myometrium and fetal membranes , in a temporal and topographically distinct manner . To address this question , we determined the localization and expression of the DB00917 receptor subtypes ( P34995 -4 ) and the PGF2alpha receptor ( P43088 ) in paired upper and lower segment myometrium , amnion , and choriodecidual samples throughout human pregnancy , with and without labor . All receptor subtypes were found throughout the muscle layers in both the upper and lower uterine segments , colocalizing with alpha smooth muscle actin . A change in intracellular localization was observed at term labor , where P34995 and P35408 were predominately associated with the nucleus . Minimal changes in the expression of the DB00917 and PGF2alpha receptor subtypes were observed with gestational age , labor , or between the upper and lower myometrial segments . Receptor expression in maternal and fetal tissues differed between the receptor subtypes ; P34995 and P35408 were predominately expressed in the fetal membranes , PTGER2 was greatest in the myometrium , whereas P43115 and P43088 were similarly expressed in the myometrium and fetal membranes . Myometrial activation through the prostaglandin receptors is perhaps more subtle and may be mediated by a balance between one or several of the prostaglandin receptor subtypes together with other known contraction associated proteins . Lack of coordination in receptor expression between the myometrium and fetal membranes may indicate different regulatory mechanisms between these tissues , or it may suggest a function for these receptors in the amnion and choriodecidua that is independent of that seen in the myometrium . A clinical trial with chimeric monoclonal antibody DB05304 and low dose interleukin-2 pulsing scheme for advanced renal cell carcinoma . PURPOSE : DB05304 is a chimeric monoclonal antibody that binds to carbonic anhydrase IX( Q16790 /MN) , which is present on greater than 95 % of RCCs of the clear cell subtype . The suggested working mechanism of DB05304 is by ADCC . Because the number of activated ADCC effector cells can be increased by a low dose interleukin-2 pulsing schedule , a multicenter study was initiated to investigate whether DB05304 combined with LD- P60568 could lead to an improved clinical outcome in patients with progressive RCC . MATERIALS AND METHODS : A total of 35 patients with progressive clear cell RCC received weekly infusions of DB05304 for 11 weeks combined with a daily LD- P60568 regimen . Patients were monitored longitudinally for ADCC capacity . Radiological assessment of metastatic lesions was performed at week 16 and regularly until disease progression . RESULTS : A durable clinical benefit was achieved in 8 of 35 patients ( 23 % ) , including 3 with a partial response and 5 with stabilization at 24 weeks or greater . Mean survival was 22 months . In general treatment was well tolerated with little toxicity . The number of effector cells increased during treatment but lytic capacity per cell did not increase . ADCC and clinical outcome did not appear to correlate . CONCLUSIONS : DB05304 combined with LD- P60568 in patients with metastatic RCC is safe and well tolerated . With a substantial clinical benefit and a median survival of 22 months in patients with metastatic RCC who have progressive disease at study entry combination therapy showed increased overall survival compared to DB05304 monotherapy . Survival was at least similar to that of currently used cytokine regimens but with a favorable toxicity profile . Effects of an Engineered Anti- P04626 Antibody chA21 on Invasion of Human Ovarian Carcinoma Cell In Vitro . OBJECTIVE : HER-2 plays an important role in the development and progression of ovarian carcinoma . A number of monoclonal antibodies ( MAbs ) and engineered antibody fragments ( such as scFvs ) against the subdomain II or IV of HER-2 extracellular domain ( O95905 ) have been developed . We investigated the effect of chA21 , an engineered anti-HER-2 antibody that bind primarily to subdomain I , on ovarian carcinoma cell invasion in vitro , and explored its possible mechanisms . METHODS : Growth inhibition of SK-OV-3 cells was assessed using a Methyl thiazolyl tetrazolium ( MTT ) assay . The invasion ability of SK-OV-3 was determined by a Transwell invasion assay . The expression of matrix metalloproteinase-2 ( P08253 ) and its tissue inhibitors ( P16035 ) was detected by immunocytochemical staining , and the expression of p38 and the phosphorylation of p38 were assayed by both immunocytochemistry and Western blot . RESULTS : After treatment with chA21 , the invasion of human ovarian cancer SK-OV-3 cells was inhibited in dose- and time-dependent manners . Simultaneously the expression of p38 , phospho-p38 , P08253 and the P08253 / P16035 ratio decreased , while P16035 expression increased . Additionally , the decrease in phospho-p38 was much greater than that of p38 . CONCLUSION : chA21 may inhibit SK-OV-3 cell invasion via the signal transduction pathway involving P08253 , P16035 , p38 and the activation of p38MAPK . Binding affinities for sulfonamide inhibitors with matrix metalloproteinase-2 using a linear response method . Due to their involvement in many pathological conditions , matrix metalloproteinases ( MMPs ) , are very attractive therapeutic targets . Our study focuses on one of them , P08253 , which is involved in tumor progression and metastasis . Recently , the solution structure of the catalytic domain of P08253 complexed with a hydroxamic acid inhibitor ( DB01630 ) was published by Feng et al . Using the Hanessian group published binding affinity data and the structure published by Feng as a basis , we have built a binding affinity model by targeting the S(2) ' pocket of the enzyme with a set of nine alpha-N-sulfonylamino hydroxamic acid derivatives . Two binding geometries of each ligand have been generated corresponding to two binding modes denoted A and B , respectively , of which the first one is targeting the S(2) ' pocket and the second one the S(1) pocket . For the binding affinity model developed for mode A the computed activities show a rmsd of 0.583 kcal/mol as compared with the experimental data , and a correlation coefficient r(2) of 0.779 , while in the case of the binding mode B a rmsd of 0.834 kcal/mol and correlation coefficient r(2) of 0.500 , respectively , were obtained . In conclusion , our data suggest a higher probability for the DB00120 (76) gated S(2) ' open form pocket to accommodate the substituent alpha versus the wide solvent exposed S(1) subsite , probability which some research groups could have overlooked due to extensive use in their calculations of non revealing S(2) ' pocket open state crystallographic structures instead of NMR ones . Attenuation of anti-tuberculosis therapy induced hepatotoxicity by Spirulina fusiformis , a candidate food supplement . Therapy using Isoniazid ( DB00951 ) and DB01045 ( Q9HBH0 ) leads to induction of hepatotoxicity in some individuals undergoing anti-tuberculosis treatment . In this study , we assessed the effect of Spirulina fusiformis on DB00951 and Q9HBH0 induced hepatotoxicity in rats compared with hepatoprotective drug Silymarin . Induction of hepatotoxicity was measured by changes in the liver marker enzymes ( aspartate transaminase , alanine transaminase , and alkaline phosphatase ) . The antioxidant status was also analyzed in liver tissue homogenate and plasma by measurement of superoxide dismutase , catalase , glutathione-S-transferase , glutathione reductase , and lipid peroxidation levels . We also aimed to study the binding and interactions of the transcription factors Pregnane X Receptor ( O75469 ) and Farnesoid X Receptor ( Q96RI1 ) with DB00951 , Q9HBH0 , and representative active compounds of Spirulina fusiformis by in silico methods . The administration of DB00951 and Q9HBH0 resulted in significant ( p < 0.05 ) decrease in the antioxidant levels and total protein levels . There was also a significant ( p < 0.05 ) increase in the levels of liver marker enzymes . Spirulina fusiformis was seen to protect the parameters from significant changes upon challenge with DB00951 and Q9HBH0 in a dose-dependent manner . This was corroborated by histological examination of the liver . The results of the in silico analyses further support the wet lab results . Therapeutic potential of highly cytotoxic natural killer cells for gastric cancer . To develop more effective therapies for patients with advanced gastric cancer , we examined the potential of ex vivo expanded natural killer ( NK ) cells . We assessed the expression of ligands for NK Group 2 Member D ( P26718 , an important NK activation molecule ) in primary tumors from 102 patients with gastric cancer by immunohistochemistry and determined their prognostic value . We then examined the in vitro and in vivo cytotoxicity of NK cells from healthy donors and patients with gastric cancer . The cytotoxicity of resting and of interleukin ( IL ) -2-activated NK cells was compared to that of NK cells expanded for 7 days by coculture with the K562-mb15-4.1BBL cell line . As a result , the expression of P26718 ligands in primary tumors was correlated with favorable presenting features and outcomes , suggesting that gastric cancer may be sensitive to NK cell cytotoxicity . Although resting NK cells showed minimal cytotoxicity against gastric cancer cells , K562-mb15-4.1BBL-expanded NK cells were highly cytotoxic and significantly more powerful than P60568 -activated NK cells . Cytotoxicity was correlated with P26718 ligand expression and could be modulated by mitogen-activated protein kinase and AKT- P19957 kinase inhibitors . The cytotoxicity of expanded NK cells against P04626 -positive gastric cancer cells could be increased by Herceptin and further augmented by DB01259 . Finally , expanded NK cells exhibited strong antitumor activity in immunodeficient mice engrafted with a gastric cancer cell line . In conclusion , gastric cancer tumors express P26718 ligands and are highly susceptible to killing by NK cells stimulated by K562-mb15-4.1BBL . These results provide a strong rationale for clinical testing of these NK cells in patients and suggest their use to augment the effects of antibody therapy . DB01259 -mediated cyclooxygenase-2 expression via epidermal growth factor receptor/ Q15717 interaction enhances the aggressiveness of triple-negative breast cancer cells . DB01259 , a dual epidermal growth factor receptor ( P00533 ) /human epidermal growth factor receptor 2 ( P04626 ) kinase inhibitor , showed clinical benefits in advanced P04626 -positive breast cancer patients . Because some triple-negative breast cancers ( TNBCs ) frequently overexpress P00533 , the antitumor activity of lapatinib in such diseases was also tested . However , the results showed a worse event-free survival rate . It remains unknown whether and how lapatinib elicits the aggressiveness of such cancer cells . In this study , our results demonstrated that lapatinib facilitated axillary and lung metastases of triple-negative MDA-MB-231 breast cancer cells without affecting their viability , leading to worse survival in orthotopic xenograft mice . The lapatinib-increased motility was attributed by the elevation of P00533 through the downregulation of microRNA-7 and by the subsequent overexpression of cyclooxygenase-2 ( P35354 ) . Strikingly , independent of its kinase activity , the elevated P00533 at least partly stabilized P35354 expression by enhancing the binding of Q15717 to P35354 mRNA . Our results suggest that lapatinib may increase the migration and invasion of MDA-MB-231 cells by upregulating P00533 and P35354 through the downregulation of microRNA-7 , providing a potential explanation for the worse clinical outcome of TNBC patients who receive lapatinib-based treatment . These findings also shed new light on the molecular mechanism of P35354 mRNA stabilization by P00533 in a kinase-independent manner . scFv-mediated delivery of truncated P55957 suppresses P04626 -positive osteosarcoma growth and metastasis . Osteosarcoma is the most common primary malignant bone tumor , with high rates of metastasis . Here , we examined the expression of human epidermal growth factor receptor-2 ( HER-2 ) in osteosarcoma cell lines with different metastatic potential , finding that the expression was correlated with metastasis of implanted tumors . We then introduced an expression vector encoding the e23sFv-PEA II-Bid O00548 -60 gene , composed of a P04626 -specific single-chain antibody fused with domain II of Pseudomonas exotoxin A ( PEA ) and the carboxy end of truncated active Bid . We demonstrated that the e23sFv-PEA II-Bid O00548 -60 molecule selectively recognized and killed P04626 -overexpressing osteosarcoma cells in vitro . Subsequently , we introduced the e23sFv-PEA II-bid O00548 -60 gene into BALB/c athymic mice bearing P04626 -positive osteosarcomas using i.m. injections of liposome-encapsulated vector . Expression of the e23sFv-PEA II-Bid O00548 -60 gene suppressed tumor growth , significantly prolonged animal survival and inhibited metastasis , thereby suggesting it may represent a competitive approach to treating P04626 /neu-positive osteosarcoma . Nearly Complete Response of Brain Metastases from P04626 Overexpressing Breast Cancer with DB01259 and DB01101 after Whole Brain Irradiation . DB00072 treatment does not prevent intracranial seeding and is largely ineffective for established central nervous system metastasis in P04626 overexpressing breast cancer patients . Combination therapy of lapatinib and capecitabine may be an effective treatment option for brain metastasis of P04626 -positive breast cancer . We report a patient with breast cancer overexpressing HER-2 where brain metastases were successfully treated with radiation and a combination of lapatinib and capecitabine . On the hepatic mechanism of HDL assembly by the O95477 /apoA-I pathway . The mechanism for the assembly of HDL with cellular lipid by O95477 and helical apolipoprotein was investigated in hepatocytes . Both HepG2 cells and mouse primary culture hepatocytes produced HDL with apolipoprotein A-I ( apoA-I ) whether endogenously synthesized or exogenously provided . DB01599 , an O95477 inactivator , inhibited these reactions , as well as the reversible binding of apoA-I to HepG2 . Primary cultured hepatocytes of O95477 -deficient mice also lacked HDL production regardless of the presence of exogenous apoA-I . HepG2 cells secreted apoA-I into the medium even when O95477 was inactivated by probucol , but it was all in a free form as HDL production was inhibited . When a lipid-free apoA-I-specific monoclonal antibody , 725-1E2 , was present in the culture medium , production of HDL was suppressed , whether with endogenous or exogenously added apoA-I , and the antibody did not influence HDL already produced by HepG2 cells . We conclude that the main mechanism for HDL assembly by endogenous apoA-I in HepG2 cells is an autocrine-like reaction in which apoA-I is secreted and then interacts with cellular O95477 to generate HDL . Novel pyrrolyllactone and pyrrolyllactam indolinones as potent cyclin-dependent kinase 2 inhibitors . P12004 -dependent kinases ( CDKs ) are essential in the control of cell cycle progression . Inhibition of CDKs represents a new approach for pharmacological intervention in the treatment of a variety of proliferative diseases , especially cancer . Based on the crystal structure of P24941 in complex with an imidazole indolinone compound 1 ( DB03428 ) , lead optimization through modeling , synthesis , and SAR studies has led to the discovery of a novel series of pyrrolyllactone and pyrrolyllactam indolinones as potent P24941 inhibitors . P35354 inhibitor treatment enhances photodynamic therapy-mediated tumor response . Photodynamic therapy ( PDT ) continues to be used in the treatment of solid tumors . Clinical results are promising , but the therapy has not been optimized , and tumor recurrences can occur . Recently , it has been shown that inhibitors of cyclooxygenase ( P36551 ) -2 can be effective in combination with conventional chemotherapy and radiation therapy . In the current study , we examined the parameters of PDT-mediated activation of P35354 expression . We also examined the tumoricidal effectiveness of combining PDT with the selective P35354 inhibitor NS-398 . PDT induced the transcriptional activation of P35354 . Prolonged expression of P35354 protein was observed in PDT-treated mouse sarcoma and carcinoma cell lines , whereas P23219 was not inducible by PDT . Prostaglandin ( PG ) E2 synthesis was also increased in PDT-treated cells , and DB00917 levels were attenuated in cells coincubated with NS-398 , indicating that PDT induced the expression of biologically active P35354 . Both porphyrin- and chlorin-based photosensitizers were able to elicit PDT-mediated P35354 expression . P35354 was also elevated in radiation-induced fibrosarcoma ( Q9HBH0 ) tumors after treatment with PDT . We also observed that systemic administration of NS-398 decreased PDT induction of both DB00917 and vascular endothelial growth factor in treated Q9HBH0 tumors . Additionally , we demonstrated that NS-398 enhanced PDT responsiveness in Q9HBH0 tumors without increasing toxicity to normal tissue . These results provide strong evidence that combination procedures involving selective P35354 inhibitors may improve the therapeutic effectiveness of PDT . P00533 pathway gene expressions and biological response of glioblastoma multiforme cell lines to erlotinib . BACKGROUND : Erlotinib , an epidermal growth factor receptor ( P00533 ) tyrosine kinase inhibitor , exerts highly variable antiproliferative effects on human glioblastoma multiforme ( GBM ) cells in vitro and in vivo . As these effects are independent of P00533 baseline expression levels , more complex genetic signatures may form the molecular basis of the erlotinib-sensitive and erlotinib-resistant GBM phenotypes . The aim of the current study was to determine which genes within the P00533 signaling pathway are candidates for mediating the cellular response of human GBM towards erlotinib . MATERIALS AND METHODS : Complementary (c)RNAs from cell lines selected to represent the sensitive , intermediately responsive and resistant phenotypes , respectively , were hybridized to CodeLink Human Whole Genome Bioarrays . RESULTS : Expression analysis of the prospectively selected 244 genes whose products constitute the P00533 signaling pathway identified five genes the expression of which significantly correlated with phenotype . Functional annotation analysis revealed one ( STATI ) and two ( Q9NWM8 , P63000 ) genes conclusively associated with sensitivity and resistance to erlotinib , respectively . Moreover , two additional genes ( P35408 , MYC ) were unexpectedly found to be associated with sensitivity . The gene expressions were confirmed by quantitative polymerase chain reaction . CONCLUSION : Five genes within the P00533 signaling pathway may modulate GBM response to erlotinib , which further emphasizes the importance of this pathway for the biology of GBM . Temperature dependence of N-methyl-D-aspartate receptor channels and N-methyl-D-aspartate receptor excitatory postsynaptic currents . N-methyl-d-aspartate ( DB01221 ) receptors ( NMDARs ) are highly expressed in the CNS and mediate the slow component of excitatory transmission . The present study was aimed at characterizing the temperature dependence of the kinetic properties of native NMDARs , with special emphasis on the deactivation of synaptic NMDARs . We used patch-clamp recordings to study synaptic NMDARs at layer II/III pyramidal neurons of the rat cortex , recombinant Q05586 / Q13224 receptors expressed in human embryonic kidney ( HEK293 ) cells , and NMDARs in cultured hippocampal neurons . We found that time constants characterizing the deactivation of NMDAR-mediated excitatory postsynaptic currents ( EPSCs ) were similar to those of the deactivation of responses to a brief application of glutamate recorded under conditions of low NMDAR desensitization ( whole-cell recording from cultured hippocampal neurons ) . In contrast , the deactivation of NMDAR-mediated responses exhibiting a high degree of desensitization ( outside-out recording ) was substantially faster than that of synaptic DB01221 receptors . The time constants characterizing the deactivation of synaptic NMDARs and native NMDARs activated by exogenous glutamate application were only weakly temperature sensitive ( Q(10)=1.7-2.2 ) , in contrast to those of recombinant Q05586 / Q13224 receptors , which are highly temperature sensitive ( Q(10)=2.7-3.7 ) . DB08954 reduced the amplitude of NMDAR-mediated EPSCs by approximately 50 % but had no effect on the time course of deactivation . Analysis of Q05586 / Q13224 responses indicated that the double exponential time course of deactivation reflects mainly agonist dissociation and receptor desensitization . We conclude that the temperature dependences of native and recombinant NMDAR are different ; in addition , we contribute to a better understanding of the molecular mechanism that controls the time course of NMDAR-mediated EPSCs .

MH_train_1362

MH_train_1362

MH_train_1362

interacts_with DB00864?

multiple_choice

O14965 promotes inflammation and tumorigenesis in mice and human gastric neoplasia . BACKGROUND & AIMS : Chronic inflammation contributes to the pathogenesis of gastric tumorigenesis . The aurora kinase A ( O14965 ) gene is frequently amplified and overexpressed in gastrointestinal cancers . We investigated the roles of O14965 in inflammation and gastric tumorigenesis . METHODS : We used quantitative real-time reverse transcription polymerase chain reaction , immunofluorescence , immunohistochemistry , luciferase reporter , immunoblot , co-immunoprecipitation , and in vitro kinase assays to analyze AGS and MKN28 gastric cancer cells . We also analyzed Tff1(-/-) mice , growth of tumor xenografts , and human tissues . RESULTS : We correlated increased expression of O14965 with increased levels of tumor necrosis factor-α and inflammation in the gastric mucosa of Tff1(-/-) mice ( r = 0.62 ; P = .0001 ) . DB05220 , an investigational small-molecule selective inhibitor of O14965 , reduced nuclear staining of nuclear factor-κB ( NF-κB ) p65 in human gastric cancer samples and mouse epithelial cells , suppressed NF-κB reporter activity , and reduced expression of NF-κB target genes that regulate inflammation and cell survival . Inhibition of O14965 also reduced growth of xenograft tumors from human gastric cancer cells in mice and reversed the development of gastric tumors in Tff1(-/-) mice . O14965 was found to regulate NF-κB activity by binding directly and phosphorylating IκBα in cells . Premalignant and malignant lesions from the gastric mucosa of patients had increased levels of O14965 protein and nuclear NF-κB , compared with healthy gastric tissue . CONCLUSIONS : In analyses of gastric cancer cell lines , human tissue samples , and mouse models , we found O14965 to be up-regulated during chronic inflammation to promote activation of NF-κB and tumorigenesis . O14965 inhibitors might be developed as therapeutic agents for gastric cancer . Novel roles of Akt and P42345 in suppressing TGF-beta/ P36897 -mediated P84022 activation . P01308 -like growth factor-I inhibits transforming growth factor-beta ( TGF-beta ) signaling by blocking activation of P84022 ( S3 ) , via a phosphatidylinositol 3-kinase ( PI3K ) /Akt-dependent pathway . Here we provide the first report that the kinase activity of Akt is necessary for its ability to suppress many TGF-beta responses , including S3 activation and induction of apoptosis . Wild-type and myristoylated Akts ( Akt(WT) and Akt(Myr) ) suppress TGF-beta-induced phospho-activation of S3 but not Q15796 ( S2 ) , whereas kinase-dead Akt1 ( Akt1K179M ) or dominant-negative PI3K enhances TGF-beta-induced phospho-activation of both S2 and S3 . Using siRNA , rapamycin ( Rap ) , and adenoviral expression for P62942 -resistant and constitutively active P36897 ( P36897 ) , we demonstrate that mammalian target of Rap ( P42345 ) mediates Akt1 suppression of phospho-activation of S3 . These and further data on Akt1-S3 binding do not support a recently proposed model that Akt blocks S3 activation through physical interaction and sequestration of S3 from TGF-beta receptors . We propose a novel model whereby Akt suppresses activation of S3 in an Akt kinase-dependent manner through P42345 , a likely route for loss of tumor suppression by TGF-beta in cancers . The specific Q14318 inhibitor N-(N',N'-dimethylcarboxamidomethyl)cycloheximide has potent neuroprotective and neurotrophic properties in brain ischemia . FK506 and FK506-derived inhibitors of the FK506-binding protein ( FKBP ) -type peptidylprolyl cis/trans-isomerases ( PPIase ) display potent neuroprotective and neuroregenerative properties in various neurodegeneration models , showing the importance of neuroimmunophilins as targets for the treatment of acute and chronic neurodegenerative diseases . However , the PPIase activity targeted by active site-directed ligands remains unknown so far . Here we show that neurotrophic FKBP ligands , such as GPI1046 and N-[methyl(ethoxycarbonyl)]cycloheximide , inhibit the calmodulin/Ca(2+) ( P62158 /Ca(2+) ) -regulated Q14318 with up to 80-fold higher affinity than P62942 . In contrast , the non-neurotrophic rapamycin inhibits Q14318 . P62158 /Ca(2+) 500-fold less affine than other neuroimmunophillins . In the context of the high expression of Q14318 in neuroblastoma cells , these data suggest that Q14318 . P62158 /Ca(2+) inhibition can mediate neurotrophic properties of FKBP ligands . The Q14318 -specific cycloheximide derivative , N-(N',N'-dimethylcarboxamidomethyl)cycloheximide ( DM-CHX ) was synthesized and used in a rat model of transient focal cerebral ischemia . Accordingly , DM-CHX caused neuronal protection as well as neural stem cell proliferation and neuronal differentiation at a dosage of 27.2 mug/kg . These effects were still dominant , if DM-CHX was applied 2-6 h post-insult . In parallel , sustained motor behavior deficits of diseased animals were improved by drug administration , revealing a potential therapeutic relevance . Thus , our results demonstrate that Q14318 inhibition by DM-CHX regulates neuronal cell death and proliferation , providing a promising strategy for the treatment of acute and/or chronic neurodegenerative diseases . A mechanism for the synergistic antimalarial action of atovaquone and proguanil . A combination of atovaquone and proguanil has been found to be quite effective in treating malaria , with little evidence of the emergence of resistance when atovaquone was used as a single agent . We have examined possible mechanisms for the synergy between these two drugs . While proguanil by itself had no effect on electron transport or mitochondrial membrane potential ( DeltaPsim ) , it significantly enhanced the ability of atovaquone to collapse DeltaPsim when used in combination . This enhancement was observed at pharmacologically achievable doses . DB01131 acted as a biguanide rather than as its metabolite cycloguanil ( a parasite dihydrofolate reductase [ P00374 ] inhibitor ) to enhance the atovaquone effect ; another P00374 inhibitor , pyrimethamine , also had no enhancing effect . DB01131 -mediated enhancement was specific for atovaquone , since the effects of other mitochondrial electron transport inhibitors , such as myxothiazole and antimycin , were not altered by inclusion of proguanil . Surprisingly , proguanil did not enhance the ability of atovaquone to inhibit mitochondrial electron transport in malaria parasites . These results suggest that proguanil in its prodrug form acts in synergy with atovaquone by lowering the effective concentration at which atovaquone collapses DeltaPsim in malaria parasites . This could explain the paradoxical success of the atovaquone-proguanil combination even in regions where proguanil alone is ineffective due to resistance . The results also suggest that the atovaquone-proguanil combination may act as a site-specific uncoupler of parasite mitochondria in a selective manner . Anti-allergic function and regulatory mechanisms of KR62980 in allergen-induced airway inflammation . The ligand-activated transcription factor , peroxisome proliferator-activated receptor ( Q07869 )gamma , and its ligands inhibit pro-inflammatory cytokine production by immune cells , thus exerting anti-inflammatory activity . As a non-thiazolidinedione PPARgamma ligand , KR62980 has anti-diabetic and anti-adipogenic activities , but its anti-inflammatory function has yet to be characterized . In this study , we investigated the functions and mechanisms of KR62980 in the activation and differentiation of P01730 + T helper ( Th ) cells by comparing its effects with those of a thiazolidinedione PPARgamma ligand , rosiglitazone . KR62980 dose-dependently and significantly suppressed TCR-triggered Th cell proliferation by suppressing P60568 /IL-2Ralpha-mediated signaling . Both KR62980 and rosiglitazone suppressed IFNgamma production in a dose-dependent manner , whereas P05112 gene expression was specifically suppressed by only KR62980 . In addition , sustained KR62980 treatment diminished Th2 cytokine production by inhibiting c-Maf expression . In vivo administration of KR62980 in a model of allergic asthma significantly attenuated eotaxin-induced eosinophil infiltration , allergic cytokine production and collagen deposition in the lung . KR62980 also decreased goblet cell hyperplasia in the airway and mucous cell metaplasia in nasal epithelium , concurrent with decreases of allergic Th2 cytokines and Q16552 in the draining lymph node . In conclusion , a novel PPARgamma ligand , KR62980 , suppresses in vitro Th2 cell differentiation and attenuates in vivo OVA-induced airway inflammation , suggesting a beneficial role for KR62980 in the treatment of allergic asthma and allergic rhinitis . Generation of Epstein-Barr virus-specific cytotoxic T lymphocytes resistant to the immunosuppressive drug tacrolimus ( FK506 ) . Adoptive transfer of autologous Epstein-Barr virus-specific cytotoxic T lymphocytes ( EBV-CTLs ) to solid organ transplant ( SOT ) recipients has been shown safe and effective for the treatment of EBV-associated posttransplantation lymphoproliferative disorders ( PTLDs ) . SOT recipients , however , require the continuous administration of immunosuppressive drugs to prevent graft rejection , and these agents may significantly limit the long-term persistence of transferred EBV-CTLs , precluding their use as prophylaxis . DB00864 ( FK506 ) is one of the most widely used immunosuppressive agents in SOT recipients , and its immunosuppressive effects are largely dependent on its interaction with the 12-kDa FK506-binding protein ( P62942 ) . We have knocked down the expression of P62942 in EBV-CTLs using a specific small interfering RNA ( siRNA ) stably expressed from a retroviral vector and found that P62942 -silenced EBV-CTLs are FK506 resistant . These cells continue to expand in the presence of the drug without measurable impairment of their antigen specificity or cytotoxic activity . We confirmed their FK506 resistance and anti-PTLD activity in vivo using a xenogenic mouse model , suggesting that the proposed strategy may be of value to enhance EBV-specific immune surveillance in patients at high risk of PTLD after transplantation . Human epidermal Langerhans ' cells are targets for the immunosuppressive macrolide tacrolimus ( FK506 ) . BACKGROUND : The immunosuppressive macrolide tacrolimus ( FK506 ) has been shown to inhibit allergic contact dermatitis in animal models as well as in human beings . More recently , successful treatment of atopic dermatitis with an ointment containing tacrolimus has been reported . OBJECTIVES : We explored the effects of this compound on epidermal Langerhans ' cells ( LCs ) , which are known to play an important pathophysiologic role in inflammatory skin diseases . METHODS : The expression of the intracellular FK506 binding protein ( P62942 ) was monitored on freshly isolated and cultured epidermal LCs . Phenotyping and functional exploration of LCs treated with different concentrations of tacrolimus and beta-methasone valerate ( betaMv ) were performed . RESULTS : P62942 is expressed in freshly isolated LCs but is lost while they are maturating into mature dendritic cells . DB00864 inhibited the expression of IL-2R ( CD25 ) and of the costimulatory molecules P33681 ( P33681 .1 ) and P25942 . Expression of MHC class I and II was also affected , whereas P42081 ( P33681 .2 ) expression was not altered . In contrast , betaMv strongly increased the expression of CD25 . Paradoxically , while decreasing P25942 and MHC class I expression , betaMv significantly increased the expression of MHC class II , P33681 , and P42081 on cultured LCs but impaired their allostimulatory activity . DB00864 was about 100 times more potent than betaMv at inhibiting LC stimulatory function . CONCLUSION : DB00864 can exert immunopharmacologic alterations on LCs , which may account , at least in part , for the therapeutic effect of this compound in eczematous skin diseases . A yeast sensor of ligand binding . We describe a biosensor that reports the binding of small-molecule ligands to proteins as changes in growth of temperature-sensitive yeast . The yeast strains lack dihydrofolate reductase ( P00374 ) and are complemented by mouse P00374 containing a ligand-binding domain inserted in a flexible loop . Yeast strains expressing two ligand-binding domain fusions , P62942 - P00374 and estrogen receptor-alpha ( ERalpha ) - P00374 , show increased growth in the presence of their corresponding ligands . We used this sensor to identify mutations in residues of ERalpha important for ligand binding , as well as mutations generally affecting protein activity or expression . We also tested the sensor against a chemical array to identify ligands that bind to P62942 or ERalpha . The ERalpha sensor was able to discriminate among estrogen analogs , showing different degrees of growth for the analogs that correlated with their relative binding affinities ( RBAs ) . This growth assay provides a simple and inexpensive method to select novel ligands and ligand-binding domains . Interaction of tacrolimus(FK506) and its metabolites with FKBP and calcineurin . DB00864 (FK506) is a strong immuno-suppressant and shows its activity through inhibiting P60568 mRNA transcription by forming pentameric complex with intracellular receptor ( FK506 binding protein 12 kDa or P62942 ) , Ca2+ , calmodulin , and calcineurin . Here , we report the binding activity to P62942 , the pentameric complex formation and Con-A response inhibiting activities of 7 metabolites . C15-demethylated metabolite(M-3) needed higher quantity to compete in Con-A assay and in pentamer formation assay , although it binds more strongly to P62942 . The result suggests that the ability to form a pentameric complex is not a two step reaction with the first binding to P62942 , but a single step reaction by components for the pentamer formation . Endometriosis is sustained by tumour necrosis factor-alpha . Endometriosis is a common gynaecological disorder causing pain , infertility , and emotional distress . Evidence presented here suggests that abnormal production of tumour necrosis factor-alpha ( P01375 ) is required for the establishment and maintenance of endometriosis and also is responsible for the associated infertility through its effect on sperm motility and function and oocyte development . DB00065 , which blocks P01375 function , could be used in the treatment of endometriosis to reverse the above effects . P22303 antagonist potentiated insulin action in fed but not fasted state . The glucose disposal effect of insulin is doubled in response to a meal . This meal-induced insulin sensitization results from insulin acting on the liver , in the presence of a permissive hepatic parasympathetic feeding signal and elevated hepatic glutathione ( DB00143 ) , to release hepatic insulin-sensitizing substance ( HISS ) , a hormone that acts selectively on skeletal muscle to stimulate insulin-mediated glucose uptake . Blockade of the parasympathetic feeding signal to the liver , either through surgical denervation or atropine-mediated antagonism of hepatic muscarinic receptors , eliminates the HISS response , resulting in HISS-dependent insulin resistance ( HDIR ) and decreasing the response to insulin by approximately 55 % in the fed state . P01308 action in Sprague-Dawley rats , as determined with a rapidly sampled , transient euglycemic clamp in response to insulin ( 50 mU/kg ) , is decreased in a dose-dependent manner by atropine . In this study , we have used the ED75 atropine-induced model of HDIR . After a submaximal dose of atropine , potentiation of the remaining parasympathetic effect with the acetylcholinesterase antagonist neostigmine significantly restored postprandial insulin sensitization in a dose-dependent manner with peak effect at 0.1 microg/kg/min . DB01400 reversed the insulin resistance induced by partial fasting and partial muscarinic inhibition ( hepatic DB00143 levels are at fed levels ) , but not that induced by surgical hepatic denervation ( DB00143 normal , no nerve signal ) or 24-h fasting ( low DB00143 ) . No potentiation of the response to insulin by neostigmine occurred in normal , fed rats . The data suggest the use of either direct or indirectly acting cholinergic agonists for the treatment of impaired postprandial insulin sensitization . Inhibition of p38alpha MAPK disrupts the pathological loop of proinflammatory factor production in the myelodysplastic syndrome bone marrow microenvironment . Myelodysplastic syndromes ( P43034 ) are common causes of ineffective hematopoiesis and cytopenias in the elderly . Various myelosuppressive and proinflammatory cytokines have been implicated in the high rates of apoptosis and hematopoietic suppression seen in P43034 . We have previously shown that p38 MAPK is overactivated in P43034 hematopoietic progenitors , which led to current clinical studies of the selective p38alpha inhibitor , DB05412 , in this disease . We now demonstrate that the myelosuppressive cytokines TNFalpha and IL-1beta are secreted by bone marrow ( BM ) cells in a p38 MAPK-dependent manner . Their secretion is stimulated by paracrine interactions between BM stromal and mononuclear cells and cytokine induction correlates with P28906 + stem cell apoptosis in an inflammation-simulated in vitro bone marrow microenvironment . Treatment with DB05412 inhibits P01375 secretion in primary P43034 bone marrow cells and protects cytogenetically normal progenitors from apoptosis ex vivo . Furthermore , p38 inhibition diminishes the expression of TNFalpha or IL-1beta-induced proinflammatory chemokines in BM stromal cells . These data indicate that p38 inhibition has anti-inflammatory effects on the bone marrow microenvironment that complements its cytoprotective effect on progenitor survival . These findings support clinical investigation of p38alpha as a potential therapeutic target in P43034 and other related diseases characterised by inflammatory bone marrow failure . Acute ethanol preexposure promotes liver regeneration after partial hepatectomy in mice by activating P05091 . It is known that chronic ethanol significantly impairs liver regeneration . However , the effect of acute ethanol exposure on liver regeneration remains largely unknown . To address this question , C57Bl6/J mice were exposed to acute ethanol ( 6 g/kg intragastrically ) for 3 days , and partial hepatectomy ( PHx ) was performed 24 h after the last dose . Surprisingly , acute ethanol preexposure promoted liver regeneration . This effect of ethanol did not correlate with changes in expression of cell cycle regulatory genes ( e.g. , cyclin D1 , P38936 , and p27 ) but did correlate with protection against the effect of PHx on indices of impaired lipid and carbohydrate metabolism . DB00898 preexposure protected against inhibition of the oxidant-sensitive mitochondrial enzyme , aconitase . The activity of aldehyde dehydrogenase 2 ( P05091 ) was significantly increased by ethanol preexposure . The effect of ethanol was blocked by inhibiting ( DB02115 ) and was mimicked by activating ( Alda-1 ) P05091 . Lipid peroxides are also substrates for P05091 ; indeed , alcohol preexposure blunted the increase in lipid peroxidation ( 4OH-nonenal adducts ) caused by PHx . Taken together , these data suggest that acute preoperative ethanol exposure " preconditions " the liver to respond more rapidly to regenerate after PHx by activating mitochondrial P05091 , which prevents oxidative stress in this compartment . Comparative transcriptional network modeling of three Q07869 -α/γ co-agonists reveals distinct metabolic gene signatures in primary human hepatocytes . AIMS : To compare the molecular and biologic signatures of a balanced dual peroxisome proliferator-activated receptor ( Q07869 ) -α/γ agonist , aleglitazar , with tesaglitazar ( a dual Q07869 -α/γ agonist ) or a combination of pioglitazone ( Pio ; Q07869 -γ agonist ) and fenofibrate ( Feno ; Q07869 -α agonist ) in human hepatocytes . METHODS AND RESULTS : Gene expression microarray profiles were obtained from primary human hepatocytes treated with EC(50)-aligned low , medium and high concentrations of the three treatments . A systems biology approach , Causal Network Modeling , was used to model the data to infer upstream molecular mechanisms that may explain the observed changes in gene expression . DB08915 , tesaglitazar and Pio/Feno each induced unique transcriptional signatures , despite comparable core Q07869 signaling . Although all treatments inferred qualitatively similar Q07869 -α signaling , aleglitazar was inferred to have greater effects on high- and low-density lipoprotein cholesterol levels than tesaglitazar and Pio/Feno , due to a greater number of gene expression changes in pathways related to high-density and low-density lipoprotein metabolism . Distinct transcriptional and biologic signatures were also inferred for stress responses , which appeared to be less affected by aleglitazar than the comparators . In particular , Pio/Feno was inferred to increase Q16236 activity , a key component of the stress response pathway , while aleglitazar had no significant effect . All treatments were inferred to decrease proliferative signaling . CONCLUSIONS : DB08915 induces transcriptional signatures related to lipid parameters and stress responses that are unique from other dual Q07869 -α/γ treatments . This may underlie observed favorable changes in lipid profiles in animal and clinical studies with aleglitazar and suggests a differentiated gene profile compared with other dual Q07869 -α/γ agonist treatments . The epidermal growth factor system in Caenorhabditis elegans . The single known epidermal growth factor-like growth factor and single epidermal growth factor receptor in Caenorhabditis elegans mediate two types of processes , each via a distinct signal transduction pathway . Several instances of cell fate specification during organogenesis require the DB01367 - Q96HU1 kinase pathway , as well as multiple nuclear factors . By contrast , appropriate myoepithelial contractions during ovulation involve IP3-mediated signal transduction . Positive modulators of the DB01367 pathway include Q8IVT5 , Q09428 -8 , phosphatase PP2A , and a zinc cation diffusion facilitator . Negative regulators of the DB01367 pathway include homologs of P22681 , P20936 -1 , ACK , and Q96HU1 kinase phosphatase , while negative regulators of the IP3 pathway are enzymes that modify IP3 . In addition to its stimulation of DB01367 activity , the P62993 homolog SEM-5 acts negatively on both signaling pathways , as does the Ack-related kinase O14965 . Pharmacology of recombinant low-voltage activated calcium channels . Several types of voltage- or ligand-activated calcium channels contribute to the excitability of neuronal cells . Low-voltage-activated ( LVA ) , T-type calcium channels are characterised by relatively negative threshold of activation and therefore they can generate low-threshold spikes , which are essential for burst firing . At least three different proteins form T-type calcium current in neurons : Ca(v)3.1 , Ca(v)3.2 and Q9P0X4 . Expression of these proteins in various brain regions is complementary . Individual channel types could be distinguished by different sensitivity towards inorganic cations . This inhibition can contribute to the toxicity of some heavy metals . Selective inhibition of T-type calcium channels by organic blockers may have clinical importance in some forms of epilepsy . DB01388 inhibits the expressed Ca(v2)3.1 , Ca(v)3.2 and Q9P0X4 channels in nanomolar concentrations with Q9P0X4 channel having lowest affinity . The sensitivity of the expressed Ca(v)3.1 channel to the antiepileptic drugs , valproate and ethosuximide , is low . Ca(v)3.1 channel is moderately sensitive to phenytoin . The Ca(v)3.2 channel is sensitive to ethosuximide , amlodipine and amiloride . All three LVA calcium channels are moderately sensitive to active metabolites of methosuximide , i.e. alpha-methyl-alpha-phenylsuccinimide . Several neuroleptics inhibit all three LVA channels in clinically relevant concentrations . All three channels are also inhibited by the endogenous cannabinoid anandamide . A high affinity peptide blocker for these Ca channels is the scorpion toxin kurtoxin which inhibits the Ca(v)3.1 and Ca(v)3.2 , but not the Q9P0X4 channel in nanomolar concentrations . DB06690 selectively inhibits the Ca(v)3.2 , but not the Ca(v)3.1 channel . The Ca(v)3.2 , but not the Ca(v)3.1 channel is potentiated by stimulation of Ca(2+)/ P62158 -dependent protein kinase . Initial trials of anti- P33681 monoclonal antibody ( DB04901 ) therapy for patients with relapsed or refractory follicular lymphoma . DB00864 ( FK506 ) increases neuronal expression of P20936 -43 and improves functional recovery after spinal cord injury in rats . DB00864 ( FK506 ) , a widely used immunosuppressant drug , has neurite-promoting activity in cultured PC12 cells and peripheral neurons . The present study investigated whether tacrolimus affects the expression of the neuronal growth-associated protein , P20936 -43 , as well as functional recovery after photothrombotic spinal cord injury in the rat . In injured animals receiving tacrolimus , the number of neurons expressing P20936 -43 mRNA and protein approximately doubled compared to that in injured animals receiving vehicle alone . This increase in P20936 -43-positive cells was paralleled by a significant improvement in neurological function evaluated by open-field and inclined plane tests . Another P62942 ligand ( V-10,367 ) had similar effects on P20936 -43 expression and functional outcome , indicating that the observed effects of tacrolimus do not involve inhibition of the phosphatase calcineurin . Thus , tacrolimus , a drug which is already approved for use in humans , as well as other P62942 ligands which do not inhibit calcineurin , could potentially enhance functional outcome after CNS injury in humans . P62942 , the 12-kDa FK506-binding protein , is a physiologic regulator of the cell cycle . P62942 , the 12-kDa FK506-binding protein , is a ubiquitous abundant protein that acts as a receptor for the immunosuppressant drug FK506 , binds tightly to intracellular calcium release channels and to the transforming growth factor beta ( TGF-beta ) type I receptor . We now demonstrate that cells from P62942 -deficient ( P62942 (-/-) ) mice manifest cell cycle arrest in G(1) phase and that these cells can be rescued by P62942 transfection . This arrest is mediated by marked augmentation of P38936 ( P38936 /CIP1) levels , which can not be further augmented by TGF-beta1 . The P38936 up-regulation and cell cycle arrest derive from the overactivity of TGF-beta receptor signaling , which is normally inhibited by P62942 . Cell cycle arrest is prevented by transfection with a dominant-negative TGF-beta receptor construct . TGF-beta receptor signaling to gene expression can be mediated by SMAD , p38 , and P29323 / Q96HU1 kinase ( extracellular signal-regulated kinase/mitogen-activated protein kinase ) pathways . SMAD signaling is down-regulated in P62942 (-/-) cells . Inhibition of P29323 / Q96HU1 kinase fails to affect P38936 up-regulation . By contrast , activated phosphorylated p38 is markedly augmented in P62942 (-/-) cells and the P38936 up-regulation is prevented by an inhibitor of p38 . Thus , P62942 is a physiologic regulator of cell cycle acting by normally down-regulating TGF-beta receptor signaling . Signaling by proinflammatory cytokines : oligomerization of TRAF2 and Q9Y4K3 is sufficient for JNK and IKK activation and target gene induction via an amino-terminal effector domain . Interleukin-1 ( IL-1 ) and tumor necrosis factor ( P01375 ) stimulate transcription factors AP-1 and NF-kappaB through activation of the Q96HU1 kinases JNK and p38 and the O15111 ( IKK ) , respectively . The P01375 and IL-1 signals are transduced through TRAF2 and Q9Y4K3 , respectively . Overexpressed TRAF2 or Q9Y4K3 activate JNK , p38 , or IKK in the absence of extracellular stimulation . By replacing the carboxy-terminal TRAF domain of TRAF2 and Q9Y4K3 with repeats of the immunophilin P62942 , we demonstrate that their effector domains are composed of their amino-terminal Zn and RING fingers . Oligomerization of the TRAF2 effector domain results in specific binding to Q13233 , a protein kinase capable of JNK , p38 , and IKK activation , and induction of P01375 and IL-1 responsive genes . P01375 also enhances the binding of native TRAF2 to Q13233 and stimulates the kinase activity of the latter . Thus , P01375 and IL-1 signaling is based on oligomerization of TRAF2 and Q9Y4K3 leading to activation of effector kinases .

MH_train_1363

MH_train_1363

MH_train_1363

interacts_with DB06271?

multiple_choice

Secreted phosphoprotein-24 kDa ( Spp24 ) attenuates P12643 -stimulated Smad 1/5 phosphorylation and alkaline phosphatase induction and was purified in a protective complex with alpha2 -Macroglobulins From Serum . Secreted phosphoprotein-24 kDa ( Spp24 ) binds cytokines of the bone morphogenetic protein/transforming growth factor-β ( BMP/TGFβ ) superfamily and is one of the most abundant serum phosphoproteins synthesized by the liver . Little is known about how Spp24 binding affects BMP signal transduction and osteoblastic differentiation or how this labile protein is transported from the liver to remote tissues , such as bone . When Spp24 was administered to W-20-17 DB05914 with rhBMP-2 , short-term Q15797 /5 phosphorylation was inhibited , intermediate-term alkaline phosphatase ( ALP ) induction was blunted , and long-term mineralization was unaffected . This supports the hypothesis that Spp24 proteolysis restricts the duration of its regulatory effects , but offers no insight into how Spp24 is transported intact from the liver to bone . When Spp24 was immunopurified from serum and subjected to native PAGE and Western blotting , a high molecular weight band of > 500 kDa was found . Under reducing SDS-PAGE , a 24 kDa band corresponding to monomeric Spp24 was liberated , suggesting that Spp24 is bound to a complex linked by disulfide bonds . However , such a complex can not be disrupted by 60 mM DB00974 under non-reducing condition or in purification buffers containing 600 mM NaCl and 0.1 % Tween-20 at pH 2.7-8.5 . LC-MS/MS analysis of affinity-purified , non-reducing SDS-PAGE separated , and trypsin digested bands showed that the Spp24 was present in a complex with three α(2) -macroglobulins ( α(2) -macroglobulin [ α(2) M ] , pregnancy zone protein [ P20742 ] and complement P01024 [ P01024 ] ) , as well as ceruloplasmin and the protease inhibitor anti-thrombin III ( P01008 ) , which may protect Spp24 from proteolysis . The polyglutamine neurodegenerative protein ataxin 3 regulates aggresome formation . The polyglutamine-containing neurodegenerative protein ataxin 3 ( P01008 ) has deubiquitylating activity and binds ubiquitin chains with a preference for chains of four or more ubiquitins . Here we characterize the deubiquitylating activity of P01008 in vitro and show it trims/edits K48-linked ubiquitin chains . P01008 also edits polyubiquitylated (125)I-lysozyme and decreases its degradation by proteasomes . Cellular studies show that endogenous P01008 colocalizes with aggresomes and preaggresome particles of the misfolded cystic fibrosis transmembrane regulator ( P13569 ) mutant CFTRDeltaF508 and associates with histone deacetylase 6 and dynein , proteins required for aggresome formation and transport of misfolded protein . Small interfering RNA knockdown of P01008 greatly reduces aggresomes formed by CFTRDeltaF508 , demonstrating a critical role of P01008 in this process . Wild-type P01008 restores aggresome formation ; however , P01008 with mutations in the active site or ubiquitin interacting motifs can not restore aggresome formation in P01008 knockdown cells . These same mutations decrease the association of P01008 and dynein . These data indicate that the deubiquitylating activity of P01008 and its ubiquitin interacting motifs as well play essential roles in CFTRDeltaF508 aggresome formation . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . Brominated cyclodipeptides from the marine sponge Geodia barretti as selective 5-HT ligands . The brominated cyclodipeptides barettin ( cyclo[(6-bromo-8-entryptophan)arginine] ) and 8,9-dihydrobarettin ( cyclo[(6-bromotryptophan)arginine] ) isolated from the marine sponge Geodia barretti have previously been shown to inhibit settlement of barnacle larvae in a dose-dependent manner in concentrations ranging from 0.5 to 25 microM . To further establish the molecular target and mode of action of these compounds , we investigated their affinity to human serotonin receptors . The tryptophan residue in the barettins resembles that of endogenous serotonin [ 5-hydroxytryptamine ] . A selection of human serotonin receptors , including representatives from all subfamilies ( 1-7 ) , were transfected into P29320 -293 cells . Barettin selectively interacted with the serotonin receptors 5- Q13049 , P28335 , and Q13639 at concentrations close to that of endogenous serotonin , with the corresponding Ki values being 1.93 , 0.34 , and 1.91 microM , respectively . 8,9-Dihydrobarettin interacted exclusively with the P28335 receptor with a Ki value of 4.63 microM ; it failed to show affinity to 5- Q13049 and Q13639 , indicating that the double bond between the tryptophan and arginine residue plays an important role in the interaction with the receptor proteins . Genetic polymorphism and activities of human lung alcohol and aldehyde dehydrogenases : implications for ethanol metabolism and cytotoxicity . DB00898 dehydrogenase ( DB00067 ) and aldehyde dehydrogenase ( ALDH ) exhibit genetic polymorphism and tissue specificity . DB00067 and ALDH isozyme phenotypes from 39 surgical Chinese lung specimens were identified by agarose isoelectric focusing . The identity of the lung beta-ADHs was further demonstrated by their characteristic pH-activity profiles for ethanol oxidation , Km values for NAD and ethanol , and inhibition by DB01213 or 1,10-phenanthroline . The beta 2 allele , coding for beta 2 polypeptide , was found to be predominant in the lung specimens studied . The DB00067 activities in the lungs with the homozygous phenotype P00325 2-2 ( exhibiting beta 2 beta 2 ) and P00325 1-1 ( exhibiting beta 1 beta 1 ) and the heterozygous phenotype P00325 P35326 ( exhibiting beta 2 beta 2 , beta 2 beta 1 , and beta 1 beta 1 ) were determined to be 999 +/- 77 , 48 +/- 17 , and 494 +/- 61 nmol/min/g tissue , respectively . Fifty-one percent of the specimens studied lacked the P05091 activity band on the isoelectric focusing gels . The activities in the lung tissues with the P05091 -active phenotype and the inactive phenotype were determined to be 30 +/- 3 and 17 +/- 1 nmol/min/g tissue , respectively . These findings indicate that human pulmonary ethanol-metabolizing activities differ significantly with respect to genetic polymorphism at both the P00325 and the P05091 loci . The results suggest that individuals with high Vmax beta 2- DB00067 and deficient in low-Km mitochondrial P05091 , accounting for approximately 45 % of the Chinese population , may end up with acetaldehyde accumulation during alcohol consumption , rendering them vulnerable to tissue injury caused by this highly reactive and toxic metabolite . P23560 and basic fibroblast growth factor downregulate DB01221 receptor function in cerebellar granule cells . Evidence has accumulated to suggest that the DB01221 glutamate receptor subtype plays an important role in neuronal degeneration evoked by hypoxia , ischemia , or trauma . Cerebellar granule cells in culture are vulnerable to DB01221 -induced neuronal excitotoxicity . In these cells , brain-derived neurotrophic factor ( P23560 ) and basic fibroblast growth factor ( P09038 ) prevent the excitotoxic effect of DB01221 . However , little is known about the molecular mechanisms underlying the protective properties of these trophic factors . Using cultured rat cerebellar granule cells , we investigated whether P23560 and P09038 prevent DB01221 toxicity by downregulating DB01221 receptor function . Western blot and RNase protection analyses were used to determine the expression of the various DB01221 receptor subunits ( Q9UHB4 , Q12879 , Q13224 , and Q14957 ) after P23560 or P09038 treatment . P09038 and P23560 elicited a time-dependent decrease in the expression of Q12879 and Q14957 subunits . Because DB01221 receptor activation leads to increased intracellular Ca2+ concentration ( [Ca2+]i ) , we studied the effect of the P23560 - and P09038 -induced reduction in Q12879 and Q14957 synthesis on the DB01221 -evoked Ca2+ responses by single-cell fura-2 fluorescence ratio imaging . P23560 and P09038 reduced the DB01221 -mediated [Ca2+]i increase with a time dependency that correlates with their ability to decrease Q12879 and Q14957 subunit expression , suggesting that these trophic factors also induce a functional downregulation of the DB01221 receptor . Because sustained [Ca2+]i is believed to be causally related to neuronal injury , we suggest that P23560 and P09038 may protect cerebellar granule cells against excitotoxicity by altering the DB01221 receptor-Ca2+ signaling via a downregulation of DB01221 receptor subunit expression . Mapping of Abll within a conserved linkage group on distal mouse chromosome 1 syntenic with human chromosome 1 using an interspecific cross . A human Abelson related gene ( P42684 ) cDNA clone was used to detect restriction fragment length polymorphisms ( RFLPs ) on mouse Southern blots . Abll was mapped to mouse chromosome 1 by analysis of segregation with other distal chromosome 1 genetic polymorphisms by using a panel of DNAs from [ ( C3H/HeJ-gld/gld x Mus spretus ) F1 x C3H/HeJ-gld/gld ] interspecific backcross mice . The data indicate the following gene order : (centromere)- P08575 -6.5 cM-Lamb- P35326 cM-Abll-2 cM-At-3 . The results extend the analysis of a large conserved linkage group spanning nearly 30 cM on distal mouse chromosome 1 syntenic with human chromosome 1q21-32 . Within this linkage group similar relative positions have been characterized in both species for P04003 , REN , P08575 , LAMB2 , P42684 , P01008 , P02652 , and P02549 . Viscoelastic sensing of conformational changes in plasminogen induced upon binding of low molecular weight compounds . P00747 is a precursor to the fibrinolytic enzyme plasmin and is known to undergo large conformational changes when subjected to low molecular lysine analogues such as tranexamic acid ( TA ) or ε-amino-n-caproic acid ( DB00513 ) . Here , we demonstrate how well-controlled surface immobilization of biotinylated plasminogen allows for monitoring of the interaction between TA and DB00513 with plasminogen . The interaction was studied by the quartz crystal microbalance with dissipation monitoring ( QCM-D ) technique as well as by surface plasmon resonance ( SPR ) based sensing . QCM-D measures changes in acoustically coupled mass ( by detection of changes in the resonance frequency of the crystal , Δf ) and is sensitive to changes in mass adsorbed on the sensor surface including how liquid medium is associated with this material . Through the dissipation factor ( i.e. , changes in the energy dissipation of the crystal oscillation , ΔD ) , QCM-D is also sensitive to the viscoelastic properties of material adsorbed to the sensor surface . Upon binding of TA or DB00513 , changes in the plasminogen structure were recorded as distinct , although small , ΔD responses which were used to determine affinity constants . By comparing native and truncated plasminogen , we conclude that the observed dissipation shifts were caused by conformational changes in the proteins leading to changes in the viscoelastic properties of the protein layer on the surface . These results demonstrate a novel application of the QCM-D technique , paving the way for a whole new approach to screening of this target for novel lead structures . DB08810 protects against ethanol-induced gastric mucosal injury in rats : role of 5-hydroxytryptamine , prostaglandins and sulfhydryl compounds . This study was designed to determine the gastroprotective properties of cinitapride ( CNT ) , a novel prokinetic benzamide derivative agonist of Q13639 and 5-HT1 receptors and 5-HT2 antagonist , on mucosal injury produced by 50 % ( v/v ) ethanol . Results were compared with those for 5-hydroxytryptamine ( 5-HT : 10 mg kg-1 ) . The possible involvements of gastric mucus secretion , endogenous prostaglandins ( PGs ) and sulfhydryl compounds ( SH ) in the protection mediated by CNT were also examined . Intraperitoneal administration of CNT ( 0.50 and 1 mg kg-1 ) , 30 min before ethanol , significantly prevented gastric ulceration and increased the hexosamine content of gastric mucus . CNT ( 1 mg kg-1 ) also produced a significant increase in gastric mucosal levels of DB00917 , but did not induce any significant changes in SH values . On the contrary , pretreatment with 5-HT worsened ethanol-induced erosions , however , did not affect gastric mucus secretion , glycoprotein content or DB00917 levels , although the non-protein SH fraction was significantly decreased . The present results demonstrate that the gastroprotective effects of CNT could be partly explained by a complex PG dependent mechanism . We suggest that 5-HT dependent mechanisms through 5-HT2 receptor blockade and 5-HT1 receptor activation could be also involved . Antithrombin III concentrate in the acute phase of thermal injury . BACKGROUND : Thermal injury disrupts homeostasis by inducing subclinical disseminated intravascular coagulation , fibrinolysis. and an acquired deficiency of Antithrombin III ( P01008 ) , a natural anticoagulant . As a result , thermally injured patients have a high incidence of hypercoagulability and thrombosis . OBJECTIVE : P01008 ( Human ) concentrate was given to a thermally injured patient to evaluate safety , and dosage requirements in this setting . DESIGN : The patient was a 40 yr old male with a 68 % total burn surface area , right femoral comminuted fracture , and P01031 - P13671 subluxation sustained in a vehicular crash . He received nine infusions of AT III ( H ) concentrate ( 100-50 u/kg ) within the first four days of injury . RESULT : The P01008 plasma level increased from 45 % on admission ( normal = 100+/-20 % ) to 120+/-25 % in the next four days . During the 64 day hospitalization , there were 11 grafting procedures with an estimated blood loss ( EBL ) /procedure : 1140 cc ; and EBL/grafted surface area ratio : 0.6 cc cm2 . The average time to healing of the meshed autograft was 6.4 days . CONCLUSION : P01008 ( H ) concentrate can be safely utilized in the acute phase of thermal injury : no excessive bleeding or prolongation of wound healing was documented . Activation of chloride secretion by isoflavone genistein in endometrial epithelial cells . BACKGROUND/AIM : DB01645 , the most active isoflavone found primarily in soybeans , alters ion transport functions in intestinal and airway epithelia . The present study aims to investigate the acute effects and mechanisms of action of genistein in immortalized porcine endometrial epithelial cells . METHODS : Ussing chamber technique was used for transepithelial electrical measurements . RESULTS : DB01645 increased short-circuit currents ( Isc ) which were inhibited by glibenclamide , P16860 , CFTRinh-172 , DIDS or bumetanide , but not amiloride . In experiments with amphotericin B-permeabilized monolayers , genistein activated the apical Cl- current and barium-sensitive basolateral K+ current while inhibiting the apical K+ current . DB01645 failed to increase the Isc in the presence of forskolin or DB07954 , but did increase the Isc in UTP . Pretreatment with genistein also abolished the increase in the Isc when induced by forskolin , DB07954 or UTP . However , Ca2+-chelating BAPTA-AM did not affect the genistein-induced increase in the Isc . The genistein-stimulated Isc was reduced by tyrosine kinase inhibitors , tyrphostin A23 or AG490 . However , vanadate , a tyrosine phosphatase inhibitor , failed to inhibit the genistein response . P03372 antagonist ICI182,780 did not alter the genistein’s action . CONCLUSION : The soy isoflavone , genistein , stimulates Cl- secretion in endometrial epithelial cells possibly via a direct activation of P13569 which appears to be modulated through a tyrosine kinase-dependent pathway . The present findings may be of benefit for the therapeutic application of genistein in the treatment of electrolyte transport disorders in the epithelia . P23560 released during neuropathic pain potentiates DB01221 receptors in primary afferent terminals . DB01221 receptors in primary afferent terminals can contribute to hyperalgesia by increasing neurotransmitter release . In rats and mice , we found that the ability of intrathecal DB01221 to induce neurokinin 1 receptor ( P25103 ) internalization ( a measure of DB05875 release ) required a previous injection of P23560 . Selective knock-down of DB01221 receptors in primary afferents decreased DB01221 -induced P25103 internalization , confirming the presynaptic location of these receptors . The effect of P23560 was mediated by tropomyosin-related kinase B ( trkB ) receptors and not p75 neurotrophin receptors ( p75(NTR) ) , because it was not produced by proBDNF and was inhibited by the trkB antagonist Q14201 -12 but not by the p75(NTR) inhibitor TAT-Pep5 . These effects are probably mediated through the truncated form of the trkB receptor as there is little expression of full-length trkB in dorsal root ganglion ( Q86YR7 ) neurons . Src family kinase inhibitors blocked the effect of P23560 , suggesting that trkB receptors promote the activation of these DB01221 receptors by Src family kinase phosphorylation . Western blots of cultured Q86YR7 neurons revealed that P23560 increased DB00135 (1472) phosphorylation of the Q13224 subunit of the DB01221 receptor , known to have a potentiating effect . Patch-clamp recordings showed that P23560 , but not proBDNF , increased DB01221 receptor currents in cultured Q86YR7 neurons . DB01221 -induced P25103 internalization was also enabled in a neuropathic pain model or by activating dorsal horn microglia with lipopolysaccharide . These effects were decreased by a P23560 scavenger , a trkB receptor antagonist and a Src family kinase inhibitor , indicating that P23560 released by microglia potentiates DB01221 receptors in primary afferents during neuropathic pain . Q13444 is an adhesion receptor for platelet P08514 -IIIa and induces platelet activation . Cell adhesion and proteolytic matrix degradation are central processes in atherosclerosis . Being a member of the family of ADAMs ( " a disintegrin and metalloproteinase " ) , metargidin ( Q13444 ) combines a metalloproteinase domain and an RGD aminoacid sequence . We studied the potential role of Q13444 as an adhesion receptor on endothelial cells and interactions between platelets and Q13444 with respect to platelet adhesion , activation and thrombus formation . Q13444 was found to be expressed on cultured endothelial cells ( HUVEC ) . Platelet adhesion to immobilized recombinant Q13444 was effectively enhanced under both static and high shear rate conditions reaching the maximum level of adhesion to fibrinogen . Consistently , platelet adhesion onto Q13444 overexpressing endothelial cells was significantly increased . Adhesion to Q13444 was reduced by blockade of P08514 -IIIa using neutralizing anti-alpha(IIb)beta3 mAbs ( DB00054 , 2G12 ) , but not by anti-alpha(v)beta3 ( LM609 ) . Soluble Q13444 binds to activated but not to resting P08514 -IIIa . Moreover , platelets adherent to Q13444 additionally attracted platelets under high shear rates indicating an initial role of platelet- Q13444 interactions for thrombus formation . Furthermore , incubation of platelets with soluble Q13444 showed a dose-dependent increase in secretion of CD62P and P29965 . Q13444 is expressed on endothelial cells and can serve as an adhesion receptor for platelets via P08514 -IIIa binding . Platelet adhesion to Q13444 leads to platelet activation , secretion and promotes thrombus formation . Thus , Q13444 may represent a novel target for antithrombotic strategies in cardiovascular pathologies . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP . Expression of ras proto-oncogenes in the Dunning R3327 rat prostatic adenocarcinoma system . Steady-state levels of c-Ha-ras mRNA were measured in eight sublines of the Dunning R3327 rat prostatic adenocarcinoma . As a control , normal dorsal prostate tissue was studied . Increased expression of c-Ha-ras is associated with tumor progression in one lineage of the Dunning R3327 system ( H to AT1 to P24752 -Lu and P24752 -Ly-Lu ) . Here ras mRNA increases as the tumor advances from androgen dependence and a high degree of differentiation to an anaplastic aneuploid phenotype with high metastatic potential . However , in the other Dunning lineage ( H to HI to HI-F to P01008 ) , expression of c-Ha-ras is variable and does not correlate with tumor progression . Immunocytochemistry showed that levels of the c-Ha-ras P38936 protein paralleled steady-state mRNA levels in variants . Transfection assays , using NIH/3T3 cells , suggested that the ras loci were not activated in the R3327 tumors . Levels of P01116 mRNA were also measured in the Dunning tumors ; these did not correlate with tumor progression in either lineage . Expression of N-ras mRNA was not detected in the Dunning tumors . Reduced fibrin deposition and intravascular thrombosis in hDAF transgenic pig hearts perfused with tirofiban . BACKGROUND : Solid organ xenograft rejection is associated with vascular injury resulting at least in part in platelet activation , and rejected xenografts invariably demonstrate intravascular thrombosis and interstitial hemorrhage . Complement activation plays a prominent role in platelet-endothelial interaction . We tested the effects of platelet P08514 /IIIa inhibitor tirofiban during perfusion of hDAF pig hearts . METHODS : Using a working-heart model , nontransgenic and hDAF pig hearts were perfused with tirofiban or human blood only . Myocardial damage was determined by hemodynamic parameters ( cardiac output , stroke work index ) and creatine phosphokinase . Further monitoring included the assessment of complement factors ( P01024 , C4 ) , platelets , fibrinogen , P01008 , and graft histology . RESULTS : Tirofiban increased cardiac output ( CO ) and stroke work index ( SWI ) of nontransgenic pig hearts and improved superior CO and SWI of hDAF pig hearts . Although perfusion time of nontransgenic pig hearts was prolonged by tirofiban ( 196+/-65 min vs. 162+/-122 min ) , a similar effect in hDAF pig hearts ( 218+/-116 min vs. 222+/-30 min ) could not be demonstrated . Tirofiban reduced consumption of P01024 and C4 independently from hDAF . Depletion of fibrinogen was equally diminished by tirofiban and hDAF ; the combination of both agents obtained no further reduction . P01008 consumption was most effectively inhibited by this combination . Intravascular fibrin deposition was reduced by tirofiban and hDAF , but particularly by the combination of the two agents . CONCLUSIONS : Improvement of heart performance and reduction of myocardial damage and intravascular thrombosis confirm a role of the P08514 /IIIa inhibitor tirofiban for the prevention of hDAF pig heart rejection and xenograft function . Targeted cytotoxic analog of luteinizing hormone-releasing hormone ( P01148 ) , AEZS-108 ( AN-152 ) , inhibits the growth of DU-145 human castration-resistant prostate cancer in vivo and in vitro through elevating P38936 and ROS levels . Management of castration-resistant prostate cancer ( CRPC ) is challenging due to lack of efficacious therapy . DB00044 -releasing hormone analogs appear to act directly on cells based on the P01148 receptors on human prostate adenocarcinoma cells . We explored anticancer activity of a cytotoxic analog of P01148 , AEZS-108 consisting of P01148 agonist linked to doxorubicin . Nude mice bearing DU-145 tumors were used to compare antitumor effects of AEZS-108 with its individual constituents or their unconjugated combination . The tumor growth inhibition of conjugate was greatest among treatment groups ( 90.5 % inhibition vs. 41 % by [D-Lys(6)] P01148 +DOX ) . The presence of P01148 receptors on DU-145 cells was confirmed by immunocytochemistry . In vitro , AEZS-108 significantly inhibited cell proliferation ( 61.2 % inhibition ) and elevated apoptosis rates ( by 46 % ) . By the detection of the inherent doxorubicin fluorescence , unconjugated doxorubicin was seen in the nucleus ; the conjugate was perinuclear and at cell membrane . Autophagy , visualized by GFP-tagged p62 reporter , was increased by AEZS-108 ( 7.9-fold vs. 5.3-fold by DOX+[D-Lys(6)] P01148 . AEZS-108 more effectively increased reactive oxygen species ( ROS , 2-fold vs. 1.4-fold by DOX+[D-Lys(6)] P01148 ) and levels of the apoptotic regulator P38936 in vivo and in vitro . We demonstrate robust inhibitory effects of the targeted cytotoxic P01148 analog AEZS-108 on P22888 positive castration-resistant prostate cancer cells . Assessment of partially deoxygenated deoxynojirimycin derivatives as glucosylceramide synthase inhibitors . Q16739 ( Q16739 ) 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 ) 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 -hydroxyl of the deoxynojirimycin core is important for Q16739 inhibition . Here we show that P13671 -OH appears of less important , which may set guidelines for the development of Q16739 inhibitors that have less affinity ( in comparison with DB00419 ) for other glycoprocessing enzymes , in particular those hydrolases that act on glucosylceramide . Effect of pulsed estrogen therapy on hemostatic markers in comparison with oral estrogen regimen in postmenopausal women . BACKGROUND/AIMS : Hormone replacement therapy ( HRT ) is associated with an increased risk of thromboembolism dependent on the type of HRT ; therefore , we compared therapy effects of intranasal with oral estrogens on coagulation and fibrinolysis markers in postmenopausal women . METHODS : A randomized study in which 29 healthy hysterectomized women received intranasal 17beta-estradiol or oral estrogens for 3 months . RESULTS : There were no significant differences in the baseline characteristics between groups . Those women receiving intranasal estradiol showed a mild increment in plasminogen activator inhibitor-1 ( P05121 -I ) ( from 6.8 +/- 3.5 to 9.6 +/- 3.9 U/ml , p < 0.01 ) ; however , fibrinogen , factor VII-tissue factor complex ( VIIa-rTF ) , antithrombin III ( P01008 ) , protein C ( PC ) activity , protein S ( PS ) activity , plasminogen ( P00747 ) , and tissue-type plasminogen activator antigen ( t-PA ) were unchanged . In contrast , oral unopposed estrogens elevated t-PA ( from 4.9 +/- 2.9 to 9.6 +/- 5.1 ng/ml , p < 0.01 ) in parallel with a decrement in P05121 -I ( from 5.2 +/- 4.0 to 2.7 +/- 1.7 U/ml , p < 0.05 ) and VIIa-rTF ( from 201.2 +/- 181.0 to 140.6 +/- 108.7 mU/ml , p < 0.05 ) . DB09222 , P01008 , PC , PS , and P00747 were unchanged . CONCLUSIONS : Nasal 17beta-estradiol had no effect on the coagulation markers , except a moderate increment in P05121 . In contrast , oral estrogens elicited a decrement in both VIIa-rTF and P05121 ; however , those changes did not surpass normal limits . Identification of antithrombin-modulating genes . Role of O95461 , a gene encoding a bifunctional glycosyltransferase , in the secretion of proteins ? The haemostatic relevance of antithrombin together with the low genetic variability of P01008 , and the high heritability of plasma levels encourage the search for modulating genes . We used a hypothesis-free approach to identify these genes , evaluating associations between plasma antithrombin and 307,984 polymorphisms in the GAIT study ( 352 individuals from 21 Spanish families ) . Despite no SNP reaching the genome wide significance threshold , we verified milder positive associations in 307 blood donors from a different cohort . This validation study suggested O95461 , a gene encoding a protein with xylosyltransferase and glucuronyltransferase activities that forms heparin-like linear polysaccharides , as a potential modulator of antithrombin based on the significant association of one SNPs , rs762057 , with anti-FXa activity , particularly after adjustment for age , sex and P01008 rs2227589 genotype , all factors influencing antithrombin levels ( p = 0.02 ) . Additional results sustained this association . O95461 silencing inHepG2 and P29320 -EBNA cells did not affect P01008 mRNA levels but significantly reduced the secretion of antithrombin with moderate intracellular retention . Milder effects were observed on α1-antitrypsin , prothrombin and transferrin . Our study suggests O95461 as the first known modifier of plasma antithrombin , and proposes a new role for O95461 in modulating extracellular secretion of certain glycoproteins . DB06693 enhances osteogenesis in murine embryonic stem cells . Embryonic stem ( ES ) cells have the capacity to differentiate into various cell types in vitro . In this study , we show that retinoic acid is important for the commitment of ES cells into osteoblasts . Culturing retinoic acid treated ES cells in the presence of the osteogenic supplements ascorbic acid and beta-glycerophosphate resulted in the expression of several osteoblast marker genes , osteocalcin , alkaline phosphatase , and osteopontin . However , there was only a slight amount of mineralized matrix secretion . Addition of bone morphogenic protein-2 or compactin , a drug of the statin family of P04035 inhibitors , resulted in a greatly enhanced formation of bone nodules . DB06693 did not modify the expression of osteogenic markers , but at the late stage of differentiation promoted an increase in P12643 expression . These results establish ES-cell derived osteogenesis as an effective model system to study the molecular mechanisms by which the statin compactin promotes osteoblastic differentiation and bone nodule formation .

MH_train_1364

MH_train_1364

MH_train_1364

interacts_with DB09053?

multiple_choice

DB09053 inhibits P11274 and NF-κB signaling and reduces tumor proliferation in tissue-resident cells of patients with CLL . Chronic lymphocytic leukemia ( CLL ) cells depend on microenvironmental factors for proliferation and survival . In particular , tissue-resident CLL cells show prominent activation of both B-cell receptor ( P11274 ) and NF-κB pathways . We evaluated the in vivo effects of ibrutinib , a Q06187 ( Q06187 ) inhibitor on tumor cell activation and proliferation in the blood , lymph node , and bone marrow of patients with CLL . Applying validated pathway-specific gene signatures , we detected a rapid and sustained downregulation of P11274 and NF-κB signaling in CLL cells from both the peripheral blood and tissue compartments during ibrutinib treatment . DB09053 reduced phosphorylation of PLCγ2 and P29323 and decreased nuclear protein expression of NF-κB p50 . DB09053 significantly decreased tumor proliferation and expression of surface activation markers Q07108 and P42081 , independent of prognostic factors such as IGHV mutational status , chromosome 17p deletion , or prior treatment history . Interestingly , stronger inhibition of P11274 signaling in lymph node resident CLL cells after one dose of ibrutinib was associated with a higher rate of nodal response at the end of cycle 2 . Together , these data validate on-target effects of Q06187 inhibition in the tissue compartments and demonstrate that ibrutinib effectively inhibits pathways that promote tumor cell activation and proliferation in vivo . This study is registered at www.clinicaltrials.gov as # NCT01500733 . Inhibitors of Q06187 and Q08881 : state of the new drugs for cancer , autoimmunity and inflammatory diseases . Q06187 and Q08881 are cytoplasmic tyrosine kinases of crucial importance for B and T cell development , with loss-of-function mutations causing X-linked agammaglobulinemia and susceptibility to severe , frequently lethal , Epstein-Barr virus infection , respectively . Over the last few years , considerable efforts have been made in order to develop small-molecule inhibitors for these kinases to treat lymphocyte malignancies , autoimmunity or allergy/hypersensitivity . The rationale is that even if complete lack of Q06187 or Q08881 during development causes severe immunodeficiency , inactivation after birth may result in a less severe phenotype . Moreover , therapy can be transient or only partially block the activity of Q06187 or Q08881 . Furthermore , a drug-induced B cell deficiency is treatable by gamma globulin substitution therapy . The newly developed Q06187 inhibitor P05154 -32765 , recently renamed DB09053 , has already entered several clinical trials for various forms of non-Hodgkin lymphoma as well as for multiple myeloma . Experimental animal studies have demonstrated highly promising treatment effects also in autoimmunity . Q08881 inhibitors are still under the early developmental phase , but it can be expected that such drugs will also become very useful . In this study , we present Q06187 and Q08881 with their signalling pathways and review the development of the corresponding inhibitors . UV-induced but P04637 independent apoptotic death in CHO. P04264 cells is promoted by M phase inhibitors . Toll-like receptor signaling is impaired in dendritic cells from patients with X-linked agammaglobulinemia . Bruton 's tyrosine kinase ( Q06187 ) , which is defective in patients with X-linked agammaglobulinemia ( XLA ) , is expressed not only in B cells but also in monocytes and dendritic cells ( DCs ) . DCs play a crucial role in the innate immune response against infections by sensing pathogens through Toll-like receptors ( TLRs ) . However , it is not known whether Q06187 deficiency in XLA might impair TLR-mediated signaling in DCs , which are susceptible to various infections . The phenotypic maturation and cytokine production mediated by TLRs were examined in monocyte-derived DC from XLA patients and normal controls . The TLR expression in DCs was analyzed by flow cytometry . TLR-mediated signaling in DCs was evaluated for the phenotypic maturation based on Q01151 expression and production of cytokines , such as P01375 , P05231 and IL-12p70 . TLR levels in DCs were similar between XLA and controls . O60603 , O00206 and Q9NYK1 /8 ligands elicited less phenotypic maturation of DCs from XLA patients than normal controls based on Q01151 expression . Stimulation with O60603 , O00206 and Q9NYK1 /8 ligands , as well as O15455 ligand , resulted in significantly lower production of P01375 , but neither P05231 nor IL-12p70 , by DCs from XLA patients in comparison to normal controls . These findings suggest that Q06187 may thus be required for TLR signaling in DCs . The impaired TLR signaling in DCs may therefore be partly responsible for the occurrence of severe infections with bacteria and some viruses in XLA patients . The sulphydryl containing P12821 inhibitor Zofenoprilat protects coronary endothelium from Doxorubicin-induced apoptosis . Pediatric and adult cancer patients , following the use of the antitumor drug Doxorubicin develop cardiotoxicity . Pharmacological protection of microvascular endothelium might produce a double benefit : ( i ) reduction of myocardial toxicity ( the primary target of Doxorubicin action ) and ( ii ) maintenance of the vascular functionality for the adequate delivery of chemotherapeutics to tumor cells . This study was aimed to evaluate the mechanisms responsible of the protective effects of the angiotensin converting enzyme inhibitor ( ACEI ) Zofenoprilat against the toxic effects exerted by Doxorubicin on coronary microvascular endothelium . We found that exposure of endothelial cells to Doxorubicin ( 0.1-1μM range ) impaired cell survival by promoting their apoptosis . P27361 /2 related p53 activation , but not reactive oxygen species , was responsible for Doxorubicin induced caspase-3 cleavage . P04637 mediated-apoptosis and impairment of survival were reverted by treatment with Zofenoprilat . The previously described PI-3K/ P29474 /endogenous fibroblast growth factor signaling was not involved in the protective effect , which , instead , could be ascribed to cystathionine gamma lyase dependent availability of H2S from Zofenoprilat . Furthermore , considering the tumor environment , the treatment of endothelial/tumor co-cultures with Zofenoprilat did not affect the antitumor efficacy of Doxorubicin . In conclusion the ACEI Zofenoprilat exerts a protective effect on Doxorubicin induced endothelial damage , without affecting its antitumor efficacy . Thus , sulfhydryl containing ACEI may be a useful therapy for Doxorubicin-induced cardiotoxicity . Partial least squares based gene expression analysis in renal failure . BACKGROUND : Preventive and therapeutic options for renal failure are still limited . Gene expression profile analysis is powerful in the identification of biological differences between end stage renal failure patients and healthy controls . Previous studies mainly used variance/regression analysis without considering various biological , environmental factors . The purpose of this study is to investigate the gene expression difference between end stage renal failure patients and healthy controls with partial least squares ( PLS ) based analysis . METHODS : With gene expression data from the Gene Expression Omnibus database , we performed PLS analysis to identify differentially expressed genes . Enrichment and network analyses were also carried out to capture the molecular signatures of renal failure . RESULTS : We acquired 573 differentially expressed genes . Pathway and Gene Ontology items enrichment analysis revealed over-representation of dysregulated genes in various biological processes . Network analysis identified seven hub genes with degrees higher than 10 , including Q86VP6 , P24941 , P04637 , Q9HCE7 , P62258 , Q07955 , and Q04206 . Proteins encoded by P24941 , P04637 , and Q04206 have been associated with the progression of renal failure in previous studies . CONCLUSIONS : Our findings shed light on expression character of renal failure patients with the hope to offer potential targets for future therapeutic studies . VIRTUAL SLIDES : The virtual slide(s) for this article can be found here : http://www.diagnosticpathology.diagnomx.eu/vs/1450799302127207 . Effects of atrial and brain natriuretic peptides upon cyclic GMP levels , potassium transport , and receptor binding in rat astrocytes . The ability of atrial natriuretic peptide ( P01160 ) and brain natriuretic peptide ( DB04899 ) to alter cyclic GMP levels and NaKCl cotransport in rat neocortical astrocytes was determined . At concentrations of 10(-9)-10(-6) M , rat ANP99-126 ( rANF ) , rat ANP102-126 ( auriculin B ) , and rat ANP103-126 ( atriopeptin III ) stimulated 6- to 100-fold increases in cyclic GMP levels . Porcine DB04899 ( pBNP ) and rat DB04899 ( rBNP ) were 20 % -90 % as effective as rANF over most of this concentration range , although 10(-6) M pBNP produced a greater effect than rANF . NaKCl cotransport as measured by bumetanide-sensitive 86Rb+ influx was not altered by exposure of astrocytes to 10(-6)M rANF , pBNP , or rBNP . Both pBNP and rBNP , as well as rat ANP103-123 ( atriopeptin I ) and des [ gl18 , ser19 , gly20 , leu21 , gly22 ] ANF4-23-NH2 ( C-ANF4-23 ) strongly competed for specific 125I-rANF binding sites in astrocyte membranes with affinities ranging from 0.03 to 0.4 nM , suggesting that virtually all binding sites measured at subnanomolar concentrations of 125I-rANF were of the P17342 ( ANF-R2 ) receptor subtype . These receptors are thought to serve a clearance function ( Maack et al. : Science 238:675-678 , 1987 ) and may be linked to a guanylate cyclase activity that is chemically and pharmacologically distinct from that coupled to P16066 ( ANF- Q96GN5 ) receptors ( Féthiere et al. : Mol Pharmacol 35:584-592 , 1989 ) . P01160 receptors on astrocytes may function in limiting the access of P01160 and DB04899 to neurons involved in body fluid and cardiovascular regulation . Multiple arrhythmic syndromes in a newborn , owing to a novel mutation in Q14524 . BACKGROUND : Mutations in the Q14524 gene have been linked to Brugada syndrome ( BrS ) , conduction disease , Long QT syndrome ( LQT3 ) , atrial fibrillation ( AF ) , and to pre- and neonatal ventricular arrhythmias . OBJECTIVE : The objective of this study is to characterize a novel mutation in Na(v)1.5 found in a newborn with fetal chaotic atrial tachycardia , post-partum intraventricular conduction delay , and QT interval prolongation . METHODS : Genomic DNA was isolated and all exons and intron borders of 15 ion-channel genes were sequenced , revealing a novel missense mutation ( Q270K ) in Q14524 . Na(v)1.5 wild type ( WT ) and Q270K were expressed in CHO- P04264 with and without the Na(v)β1 subunit . Results . Patch-clamp analysis showed ∼40 % reduction in peak sodium channel current ( I(Na) ) density for Q270K compared with WT . Fast and slow decay of I(Na) were significantly slower in Q270K . Steady-state activation and inactivation of Q270K channels were shifted to positive potentials , and window current was increased . The tetrodotoxin-sensitive late I(Na) was increased almost 3-fold compared with WT channels . DB00243 reduced late I(Na) in WT and Q270K channels , while exerting minimal effects on peak I(Na) . CONCLUSION : The Q270K mutation in Q14524 reduces peak I(Na) while augmenting late I(Na) , and may thus underlie the development of atrial tachycardia , intraventricular conduction delay , and QT interval prolongation in an infant . miR-7 inhibits glioblastoma growth by simultaneously interfering with the PI3K/ Q06187 and Raf/MEK/ P29323 pathways . P00533 ( P00533 ) signaling regulates glioblastoma cell proliferation , survival , migration and invasion and plays a key role in tumor progression . We show that microRNA-7 ( miR-7 ) is a common regulator of the phosphoinositide-3-kinase ( PI3K ) / Q06187 and Raf/mitogen-activated protein kinase kinase ( MEK ) /extracellular signal-regulated kinase ( P29323 ) pathways , both of which are launched by P00533 through its two direct targets , the transcription factors PI3K and P04049 , respectively . Enforced expression of miR-7 markedly decreased expression of PI3K , phosphorylated Akt , P04049 , phosphorylated Q02750 /2 , and cyclin D1 , as well as slightly reduced expression of P00533 . Forced expression of PI3K or P04049 transcripts lacking the 3'-untranslated region ( 3'-UTR ) partially reversed the effects of miR-7 on cell growth inhibition and cell cycle arrest in glioma cells . Additionally , transient expression of miR-7 in glioblastoma cells strongly inhibited in vivo glioblastoma xenograft growth . We conclude that miR-7 is a potential tumor suppressor in glioblastoma that acts by targeting multiple oncogenes related to the downstream pathway of P00533 and may serve as a novel therapeutic target for malignant gliomas . The human organic anion transporter 2 gene is transactivated by hepatocyte nuclear factor-4 alpha and suppressed by bile acids . The human organic anion transporter 2 ( Q9Y694 , Q9Y694 ) mediates the sodium-independent uptake of numerous drugs , including cephalosporins , salicylates , dicarboxylates , and prostaglandins , and is mainly expressed in hepatocytes . Because the regulation of Q9Y694 expression is poorly understood , we characterized cis-acting elements in the 5'-flanking region that regulate Q9Y694 transcription . A consensus binding motif for the hepatocyte nuclear factor-4 alpha ( HNF-4 alpha ) , arranged as a direct repeat ( DR ) -1 , is located at nucleotides -329/-317 relative to the transcription initiation site . This element specifically binds HNF-4 alpha in electrophoretic mobility shift assays . A luciferase-linked Q9Y694 promoter fragment containing the HNF-4 alpha binding site was transactivated upon cotransfection of an HNF-4 alpha expression vector in Huh7 cells , whereas site-directed mutagenesis of the P04229 element abolished activation by HNF-4 alpha . Short interfering RNAs inhibiting endogenous HNF-4 alpha expression markedly reduced endogenous expression of Q9Y694 in Huh7 cells . Because HNF-4 alpha is a known target for bile acid-mediated repression of gene transcription , we studied whether chenodeoxycholic acid ( DB06777 ) suppresses Q9Y694 gene expression by inhibiting HNF-4 alpha-mediated transactivation . Treatment of Huh7 cells with DB06777 or the synthetic farnesoid X receptor ( Q96RI1 ) agonist GW4064 decreased mRNA and protein levels and also nuclear binding activity of HNF-4 alpha . The Q96RI1 -inducible transcriptional repressor small heterodimer partner inhibited transactivation of Q9Y694 promoter constructs and of endogenous Q9Y694 expression by HNF-4 alpha . We conclude that the Q9Y694 gene is critically dependent on HNF-4 alpha and that bile acids repress the Q9Y694 gene by inhibiting HNF-4 alpha . Hepatic uptake of Q9Y694 substrates may thus be decreased in disease conditions associated with elevated intracellular levels of bile acids . " Accomodated " pig endothelial cells promote nitric oxide-dependent Th-2 cytokine responses from human T cells . BACKGROUND : Cardiac and renal allo- and xenografts can become naturally resistant to vascular rejection . Understanding this process of " accommodation " would enhance our understanding of vascular inflammatory responses and have implications for immune manipulation and tolerance induction . A feature of these grafts is infiltration by leukocytes secreting a Th-2 pattern of cytokines . METHODS : HLA- P04229 -transfected , immortalized porcine endothelial cells ( IPEC ) were incubated with polyclonal human immunoglobulin G ( IgG ) for 6 days before incubation with purified human P01730 + T cells . RESULTS : IgG-incubated IPEC stimulated a normal proliferative response from alloreactive T cells . However , interferon ( IFN ) -gamma levels were significantly reduced , whereas interleukin ( IL ) -5 and P22301 were maintained at levels equivalent to those stimulated by control IPEC . Cognate interaction between T cells and IPEC was not required for this effect , because IgG-incubated , MHC-class II-negative IPEC caused reduced P01579 secretion during a response to human Epstein-Barr virus-transformed B cells . Experiments with the nitric oxide ( NO ) donor , (z)-1-2- [ 2-Aminoethyl ) -N-(2-ammonioethyl)amino ] diazen-1-ium-1,2-diolate ( DETA-NO ) , and the NO synthase inhibitor , NG-monomethyl-L-arginine.monoacetate ( L-NMMA ) , showed that NO released by the IgG-incubated IPEC was actively involved in the development of this phenotype . CONCLUSIONS : These data suggest a novel , IgG-mediated , NO-dependent mechanism by which endothelial cells ( EC ) influence T cell responsiveness and that the Th-2 cytokine skewing seen in " accommodated " grafts may be a secondary phenomenon , resulting from the T-EC interactions . Antisense oligonucleotides : target validation and development of systemically delivered therapeutic nanoparticles . Antisense oligonucleotides ( ASO ) against specific molecular targets ( e.g. , Bcl-2 and P04049 ) are important reagents in cancer biology and therapy . Phosphorothioate modification of the ASO backbone has resulted in an increased stability of ASO in vivo without compromising , in general , their target selectivity . Although the power of antisense technology remains unsurpassed , dose-limiting side effects of modified ASO and inadequate penetration into the tumor tissue have necessitated further improvements in ASO chemistry and delivery systems . Oligonucleotide delivery systems may increase stability of the unmodified or minimally modified ASO in plasma , enhance uptake of ASO by tumor tissue , and offer an improved therapy response . Here , we provide an overview of ASO design and in vivo delivery systems , and focus on preclinical validation of a liposomal nanoparticle containing minimally modified raf antisense oligodeoxynucleotide ( DB04973 ) . Intact rafAON ( 15-mer ) is present in plasma and in normal and tumor tissues of athymic mice systemically treated with DB04973 . P04049 expression is decreased in normal and tumor tissues of DB04973 -treated mice . Therapeutic benefit of a combination of DB04973 and radiation or an anticancer drug exceeds radiation or drug alone against human prostate , breast , and pancreatic tumors grown in athymic mice . Further improvements in ASO chemistry and nanoparticles are promising avenues in antisense therapy of cancer . DB01780 signaling reveals 14-3-3 protein function as a novel step in left-right patterning during amphibian embryogenesis . To gain insight into the molecular mechanisms underlying the control of morphogenetic signals by H+ flux during embryogenesis , we tested DB01780 -A ( FC ) , a compound produced by the fungus Fusicoccum amygdali Del . In plant cells , FC complexes with 14-3-3 proteins to activate H+ pumping across the plasma membrane . It has long been thought that FC acts on higher plants only ; here , we show that exposing frog embryos to FC during early development specifically results in randomization of the asymmetry of the left-right ( LR ) axis ( heterotaxia ) . Biochemical and molecular-genetic evidence is presented that 14-3-3-family proteins are an obligate component of Xenopus FC receptors and that perturbation of 14-3-3 protein function results in heterotaxia . The subcellular localization of 14-3-3 mRNAs and proteins reveals novel cytoplasmic destinations , and a left-right asymmetry at the first cell division . Using gain-of-function and loss-of-function experiments , we show that P62258 protein is likely to be an endogenous and extremely early aspect of LR patterning . These data highlight a striking conservation of signaling pathways across kingdoms , suggest common mechanisms of polarity establishment between C. elegans and vertebrate embryos , and uncover a novel entry point into the pathway of left-right asymmetry determination . Novel phenolic antioxidants as multifunctional inhibitors of inducible P19320 expression for use in atherosclerosis . A series of novel phenolic compounds has been discovered as potent inhibitors of P01375 -inducible expression of vascular cell adhesion molecule-1 ( P19320 ) with concurrent antioxidant and lipid-modulating properties . Optimization of these multifunctional agents led to the identification of 3a ( DB05399 ) as a clinical candidate with demonstrated efficacies in animal models of atherosclerosis and hyperlipidemia . Potent and selective inhibition of human nitric oxide synthases . Inhibition by non-amino acid isothioureas . DB02539 was a potent competitive inhibitor of human nitric oxide synthase ( NOS ) , with Ki values of 17 , 36 , and 29 nM for the inducible ( i ) , endothelial ( e ) , and neuronal ( n ) isozymes , respectively . Unlike some potent inhibitors of NOS , no time dependence was observed . DB02539 was not a detectable substrate for P29474 . DB02539 was also a potent inhibitor of mouse P35228 ( Ki value of 5.2 nM ) , and its binding perturbed the spectrum of P35228 consistent with its altering the environment of the bound heme . The optimum binding of S-ethyl- and S-isopropylisothiourea relative to 70 other analogs suggested that these alkyl substitutions fit into a small hydrophobic pocket . Most isothioureas were 2-6-fold selective for the human P35228 ( Ki for P35228 versus Ki for P29474 ) , with one being 19-fold selective . The cyclized mimics of S-ethylisothiourea , 2-NH2-thiazoline , and 2-NH2-thiazole , were also competitive inhibitors of human NOS . A third structural class of inhibitors , bisisothioureas , were , in general , the most selective in their inhibition of human P35228 . S,S'-(1,3-Phenylenebis(1,2-ethanediyl))bisisothiourea was 190-fold selective ( Ki value of 0.047 microM against P35228 versus 9.0 microM against P29474 ) . These results demonstrate that potent and selective inhibition of human NOS isozymes is achievable . P40763 repressed Q93009 expression is crucial for colon cancer development . P05231 ( P05231 ) induced P40763 activation is viewed as crucial for multiple tumor growth and metastasis , including colon cancer . However , the molecular mechanisms remain largely unexplored . Here , we show that expression of ubiquitin-specific protease 7 ( Q93009 ) , a deubiquitylating enzyme , is decreased in P40763 -positive tumors . P05231 administration or transfection of a constitutively activated P40763 in SW480 cells also repressed USP mRNA expression . Using luciferase reporter and ChIP assay , we found that P40763 bound to the promoter region of Q93009 and inhibited its activity through recruiting Q13547 . As a result of the decline of Q93009 expression , endogenous P04637 protein level was decreased . Thus , our results suggest a previously unknown P40763 - Q93009 - P04637 molecular network controlling colon cancer development . [ DB09053 : A new drug of B-cell malignancies ] . DB09053 ( Imbruvica® ) is a first-in-class , orally administered once-daily , that inhibits B-cell antigen receptor signaling downstream of Bruton 's tyrosine kinase ( Q06187 ) . DB09053 has been approved in USA in February 2014 and in France in October 2014 for the treatment of patients with relapsed/refractory mantle cell lymphoma ( Q8WXI8 ) or chronic lymphocytic leukaemia ( CLL ) and for the treatment of patients with CLL and a chromosome 17 deletion ( del 17p ) or P04637 mutation . In clinical studies , ibrutinib induced an impressive overall response rate ( 68 % ) in patients with relapsed/refractory Q8WXI8 ( phase II study ) . In CLL , ibrutinib has shown to significantly improve progression-free survival , response rate and overall survival in patients with relapsed/refractory CLL , including in those with del 17p . DB09053 had an acceptable tolerability profile . Less than 10 % of patients discontinued their treatment because of adverse events . Results are pending in other B-cell lymphomas subtypes such as in diffuse large B-cell lymphoma and in follicular lymphoma . An approval extension has already been enregistered for Waldenström disease in USA in January 2015 . Given its efficacy and tolerability , ibrutinib is an emerging treatment option for patients with B-cell malignancies . DB09053 treatment ameliorates murine chronic graft-versus-host disease . Chronic graft-versus-host disease ( cGVHD ) is a life-threatening impediment to allogeneic hematopoietic stem cell transplantation , and current therapies do not completely prevent and/or treat cGVHD . P01730 + T cells and B cells mediate cGVHD ; therefore , targeting these populations may inhibit cGVHD pathogenesis . DB09053 is an FDA-approved irreversible inhibitor of Bruton 's tyrosine kinase ( Q06187 ) and P60568 inducible T cell kinase ( Q08881 ) that targets Th2 cells and B cells and produces durable remissions in B cell malignancies with minimal toxicity . Here , we evaluated whether ibrutinib could reverse established cGVHD in 2 complementary murine models , a model interrogating T cell-driven sclerodermatous cGVHD and an alloantibody-driven multiorgan system cGVHD model that induces bronchiolar obliterans ( BO ) . In the T cell-mediated sclerodermatous cGVHD model , ibrutinib treatment delayed progression , improved survival , and ameliorated clinical and pathological manifestations . In the alloantibody-driven cGVHD model , ibrutinib treatment restored pulmonary function and reduced germinal center reactions and tissue immunoglobulin deposition . Animals lacking Q06187 and Q08881 did not develop cGVHD , indicating that these molecules are critical to cGVHD development . Furthermore , ibrutinib treatment reduced activation of T and B cells from patients with active cGVHD . Our data demonstrate that B cells and T cells drive cGVHD and suggest that ibrutinib has potential as a therapeutic agent , warranting consideration for cGVHD clinical trials . DB09210 defines a new binding site for allosteric modulators of alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid ( AMPA ) receptors . Glutamate receptors are the most prevalent excitatory neurotransmitter receptors in the vertebrate central nervous system and are important potential drug targets for cognitive enhancement and the treatment of schizophrenia . Allosteric modulators of AMPA receptors promote dimerization by binding to a dimer interface and reducing desensitization and deactivation . The pyrrolidine allosteric modulators , piracetam and aniracetam , were among the first of this class of drugs to be discovered . We have determined the structure of the ligand binding domain of the AMPA receptor subtypes P42262 and P42263 with piracetam and a corresponding structure of P42263 with aniracetam . Both drugs bind to P42262 and P42263 in a very similar manner , suggesting little subunit specificity . However , the binding sites for piracetam and aniracetam differ considerably . DB04599 binds to a symmetrical site at the center of the dimer interface . DB09210 binds to multiple sites along the dimer interface with low occupation , one of which is a unique binding site for potential allosteric modulators . This new site may be of importance in the design of new allosteric regulators . Mapping of 13 horse genes by fluorescence in-situ hybridization ( Q5TCZ1 ) and somatic cell hybrid analysis . We report fluorescence in-situ hybridization ( Q5TCZ1 ) and somatic cell hybrid mapping data for 13 different horse genes ( P01160 , P06729 , P10909 , P54108 , P05093 , P02679 , P18510 , P22301 , P45452 , PRM1 , P35354 , P01375 and P04637 ) . Primers for PCR amplification of intronic or untranslated regions were designed from horse-specific DNA or mRNA sequences in GenBank . Two different horse bacterial artificial chromosome ( BAC ) libraries were screened with PCR for clones containing these 13 Type I loci , nine of which were found in the libraries . BAC clones were used as probes in dual colour Q5TCZ1 to confirm their precise chromosomal origin . The remaining four genes were mapped in a somatic cell hybrid panel . All chromosomal assignments except one were in agreement with human-horse ZOO- Q5TCZ1 data and revealed new and more detailed information on the equine comparative map . P10909 was mapped by synteny to ECA2 while human-horse ZOO- Q5TCZ1 data predicted that P10909 would be located on ECA9 . The assignment of P18510 permitted analysis of gene order conservation between HSA2 and ECA15 , which identified that an event of inversion had occurred during the evolution of these two homologous chromosomes . Short and long access to cocaine self-administration activates tyrosine phosphatase P54829 and attenuates GluN expression but differentially regulates GluA expression in the prefrontal cortex . RATIONALE : Dephosphorylation of extracellular signal-regulated kinase ( P29323 ) and cyclic AMP response element binding protein ( CREB ) in the dorsomedial prefrontal cortex ( dmPFC ) at the end of short access ( ShA ) cocaine self-administration is implicated in cocaine seeking . However , what receptors and phosphatases mediate this effect and whether P29323 /CREB and related phospho-proteins in the dmPFC react similarly during early withdrawal from long access ( LgA ) cocaine self-administration are unknown . OBJECTIVES : The effects of ShA vs. LgA cocaine self-administration on the phosphorylation of protein phosphatase 2A ( PP2A ) and striatal-enriched protein tyrosine phosphatase ( P54829 ) , as well as GluN and GluA receptor subtype expression in the dmPFC during early withdrawal , were compared . METHODS : Rats self-administered cocaine or received saline during 2- or 6-h daily sessions for 10-11 days . Two hours after the final session , the dmPFC was dissected out and processed for immunoblotting . RESULTS : Similar to previous findings after ShA cocaine , phospho- P29323 and phospho-CREB in the dmPFC were decreased after LgA cocaine . Cocaine elevated phospho-PP2A ( deactivation ) and decreased phospho- P54829 ( activation ) in both ShA and LgA cocaine rats . Q05586 , Q13224 , and phospho- Q13224 Tyr1472 in the dmPFC were decreased after ShA and LgA cocaine . Further , a significant reduction of P42262 , P42261 , and phospho- P42261 Ser845 was found only in LgA rats . CONCLUSIONS : Activation of phospho- P54829 may underlie P29323 and CREB dephosphorylation in the dmPFC as well as internalization and degradation of GluN complexes during early withdrawal from both ShA and LgA cocaine self-administration , whereas differential alteration of AMPA receptor subunits after ShA and LgA cocaine self-administration depends on cocaine intake . Discovery of ( 2E ) -3-{2-butyl-1-[2-(diethylamino)ethyl]-1H-benzimidazol-5-yl}-N-hydroxyacrylamide ( DB05223 ) , an orally active histone deacetylase inhibitor with a superior preclinical profile . A series of 3-(1,2-disubstituted-1H-benzimidazol-5-yl)-N-hydroxyacrylamides ( 1 ) were designed and synthesized as HDAC inhibitors . Extensive SARs have been established for in vitro potency ( Q13547 enzyme and COLO 205 cellular IC(50) ) , liver microsomal stability ( t(1/2) ) , cytochrome P450 inhibitory ( 3A4 IC(50) ) , and clogP , among others . These parameters were fine-tuned by carefully adjusting the substituents at positions 1 and 2 of the benzimidazole ring . After comprehensive in vitro and in vivo profiling of the selected compounds , DB05223 ( 3 ) was identified as a preclinical development candidate . 3 is a potent pan-HDAC inhibitor with excellent druglike properties , is highly efficacious in in vivo tumor models ( HCT-116 , PC-3 , A2780 , MV4-11 , Ramos ) , and has high and dose-proportional oral exposures and very good ADME , safety , and pharmaceutical properties . When orally dosed to tumor-bearing mice , 3 is enriched in tumor tissue which may contribute to its potent antitumor activity and prolonged duration of action . 3 is currently being tested in phase I and phase II clinical trials .

MH_train_1365

MH_train_1365

MH_train_1365

interacts_with DB06589?

multiple_choice

DB00139 dehydrogenase gene mutations are strongly associated with paraganglioma of the organ of Zuckerkandl . Organ of Zuckerkandl paragangliomas ( PGLs ) are rare neuroendocrine tumors that are derived from chromaffin cells located around the origin of the inferior mesenteric artery extending to the level of the aortic bifurcation . Mutations in the genes encoding succinate dehydrogenase subunits ( SDH ) B , C , and D ( SDHx ) have been associated with PGLs , but their contribution to PGLs of the organ of Zuckerkandl PGLs is not known . We aimed to describe the clinical presentation of patients with PGLs of the organ of Zuckerkandl and investigate the prevalence of SDHx mutations and other genetic defects among them . The clinical characteristics of 14 patients with PGL of the organ of Zuckerkandl were analyzed retrospectively ; their DNA was tested for SDHx mutations and deletions . Eleven out of 14 ( 79 % ) patients with PGLs of the organ of Zuckerkandl were found to have mutations in the P21912 ( 9 ) or O14521 ( 2 ) genes ; one patient was found to have the Carney-Stratakis syndrome ( CSS ) , and his PGL was discovered during surgery for gastrointestinal stromal tumor . Our results show that SDHx mutations are prevalent in pediatric and adult PGLs of the organ of Zuckerkandl . Patients with PGLs of the organ of Zuckerkandl should be screened for SDHx mutations and the CSS ; in addition , asymptomatic carriers of an SDHx mutation among the relatives of affected patients may benefit from tumor screening for early PGL detection . New analogs of vitamin D3 . Calcitriol , the most active metabolite of vitamin D , controls parathyroid gland growth and suppresses the synthesis and secretion of parathyroid hormone ( PTH ) . However , because of its potent effects on intestinal calcium absorption and bone mobilization , calcitriol treatment can induce hypercalcemia , often precluding its use at therapeutic doses . Hyperphosphatemia is also a persistent problem among patients undergoing chronic hemodialysis and can be aggravated by therapeutic doses of calcitriol . Several pharmaceutical companies were able to modify the side-chain of the 1,25(OH)2D3 , allowing some of these new analogs to retain the action on the parathyroid glands while decreasing their hypercalcemic and hyperphosphatemic effects . The structure-activity relationship for ligand-mediated transcriptional regulation has been studied in detail . In some analogs the serum binding protein ( DBP ) plays a key role in determining the pharmacokinetics of the vitamin D compound . The affinity to DBP for 22-oxacalcitriol ( O75051 ) , an analog of calcitriol for the treatment of secondary hyperparathryoidism , is approximately 300-400 times lower than that of calcitriol and the analog is rapidly cleared from the circulation . The mechanisms for the selectivity of 19-nor-1,25(OH)2D2 ( paricalcitol ) ( DB00910 ) another analog of calcitriol , is clearly different from O75051 . Although the mechanisms of action is not completely known , it does appear that paricalcitol down-regulates the P11473 in the intestine . It is likely that the unique biological profiles of vitamin D analogs in vivo are due to multiple mechanisms . Understanding the molecular basis of the analog selectivity will not only provide an explanation for their unique actions but allow intelligent design of more effective analogs in the future . Order of genes on human chromosome 5q with respect to 5q interstitial deletions . Using ( a ) somatic cell hybrids retaining partial chromosome 5 and ( b ) clinical samples from patients with acquired deletions of the long arm of chromosome 5 , combined with chromosome 5-linked DNA probes , some of which exhibited RFLPs , we have determined the order of a series of genes on chromosome 5 . The order established is 5pter ---- MLVI-2 ---- cen ---- P07686 ---- P00374 ---- Pi227- --- cp12.6 ---- ( P05113 , P05112 ) ---- P08700 ---- P04141 ---- P05230 ---- ( P07333 , P09619 ) ---- ( treC,ADRBR ) ---- ( Q5SW96 - Q13585 , P09603 ) ---- qter . The suggested order and orientation for the closely linked P08700 / P04141 gene pair is cen ---- 5' P08700 3' ---- 5' P04141 3' ---- qter , on the basis of analysis of the P04141 rearrangement in HL60 DNA . The map position of the GRL locus , which was consistent with both somatic cell hybrid and 5q- analyses , was telomeric to P04141 and centromeric to P07333 / P09619 , near P05230 . Long-range restriction-enzyme analysis of 5q- DNAs did not detect rearrangements of 5q-linked probes except in HL60 DNA , but it did reveal putative long-range RFLPs of several loci . RFLPs for GRL , Pi227 , cp12.6 , P08700 , and P07333 can detect deletions in bone marrow and in leukemia cells from patients with acquired 5q deletions . DB00134 recycling as a target for antiprotozoal drug development . The development of new and effective ontiprotozool drugs has been difficult because of the close metabolic relationship between protozoa and mammalian cells . In this article , Michael Riscoe , Al Ferro and john Fitchen present their hypothesis for chemotherapeutic exploitation of methylthioribose ( Q99707 ) kinase , an enzyme critical to methionine salvage in certain protozoa . They propose that analogues of Q99707 if properly designed , would be converted to toxic products in organisms that contain Q99707 kinase but not in mammalian cells , which lack this enzyme . Synergistic inhibition of breast cancer cell lines with a dual inhibitor of P00533 -HER-2/neu and a Bcl-2 inhibitor . The epidermal growth factor receptor ( P00533 ) ( ErbB1 ) and HER-2/neu ( ErbB2 ) are members of the ErbB family of receptor tyrosine kinases . These receptors are overexpressed in a variety of human tumors and overexpression generally correlates with poor prognosis and decreased survival . DB01259 , a reversible inhibitor of both P00533 and HER-2/neu , has shown some success in achieving clinical responses in heavily pretreated advanced cancer patients . GW2974 is a reversible dual inhibitor similar to lapatinib , but GW2974 was not progressed to clinical trials due to pharmacokinetic issues . Bcl-2 , an anti-apoptotic protein , is also overexpressed in a number of human tumors . Bcl-2 inhibitors induce apoptosis and sensitize cancer cells to other therapies . The purpose of this study was to assess the effects of combining ErbB and Bcl-2 inhibitors on the growth of human breast cancer cell lines . P00533 /HER-2/neu tyrosine kinase inhibitors ( lapatinib and GW2974 ) were combined with Bcl-2 inhibitors ( HA14-1 or GX15-070 ) and the anti-proliferative effects were determined by the MTT tetrazolium dye assay . Combinations were tested in MCF-7 human breast cancer cells , a HER-2/neu transfected MCF-7 cell line ( MCF/18 ) , and a tamoxifen-resistant MCF-7 cell line ( Q99707 -3 ) . A synergistic inhibitory effect was observed with the combination of inhibitors of P00533 -HER-2/neu ( lapatinib or GW2974 ) and Bcl-2 ( GX15-070 or HA14-1 ) on the growth of the MCF-7 , MCF/18 , and Q99707 -3 human breast cancer cell lines . This study suggests that simultaneously blocking the ErbB family of receptor tyrosine kinases and Bcl-2 family of proteins may be a benefit to breast cancer patients . Contributions of molecular analysis to the diagnosis and treatment of gastrointestinal neoplasms . This review discusses the role of molecular analysis in the diagnosis and treatment of gastrointestinal ( GI ) neoplasms . It is divided into 3 sections . The first section describes clinical applications of 11 immunohistochemical stains ( p53 , P04626 , P10721 , P21912 , Q13485 , beta-catenin , P07148 , P40692 , P54278 , P43246 , and P52701 ) , the results of which directly reflect underlying genetic or epigenetic events . These applications are mainly diagnostic but in a few instances are predictive . Germline mutation testing is a diagnostic cornerstone in the hereditary cancer predisposition syndromes ( HCPSs ) . Section two will describe the genotype and phenotype of 8 HCPSs presenting in the GI tract . Where available , guidelines based on evidence and/or expert opinion as to whom to test are presented . With our ever-expanding knowledge of the molecular genetic basis of cancer and an increasingly " biologic-oriented " therapeutic armamentarium , pathologists play a vital role in directing molecular-based predictive testing . The final section will discuss the 4 most mature examples in the GI tract : ( 1 ) P04626 testing to select patients with advanced gastroesophageal adenocarcinoma for anti- P04626 therapy , ( 2 ) P10721 and P16234 mutation analysis to direct tyrosine kinase inhibitor therapy in gastrointestinal stromal tumor , ( 3 ) DNA mismatch repair function testing to determine the applicability of adjuvant chemotherapy in patients with stage II colorectal cancer ( CRC ) , and ( 4 ) P01116 mutation analysis and related testing to determine the appropriateness of anti- P00533 monoclonal antibody therapy in patients with metastatic CRC . The identification and characterization of breast cancer CTCs competent for brain metastasis . Brain metastatic breast cancer ( BMBC ) is uniformly fatal and increasing in frequency . Despite its devastating outcome , mechanisms causing BMBC remain largely unknown . The mechanisms that implicate circulating tumor cells ( CTCs ) in metastatic disease , notably in BMBC , remain elusive . We characterize CTCs isolated from peripheral blood mononuclear cells of patients with breast cancer and also develop CTC lines from three of these patients . In epithelial cell adhesion molecule ( EpCAM ) -negative CTCs , we identified a potential signature of brain metastasis comprising " brain metastasis selected markers (BMSMs) " P04626 + / P00533 + / Q9Y251 + / Notch1+ . These CTCs , which are not captured by the CellSearch platform because of their EpCAM negativity , were analyzed for cell invasiveness and metastatic competency in vivo . CTC lines expressing the BMSM signature were highly invasive and capable of generating brain and lung metastases when xenografted in nude mice . Notably , increased brain metastatic capabilities , frequency , and quantitation were detected in EpCAM- CTCs overexpressing the BMSM signature . The presence of proteins of the BMSM CTC signature was also detected in the metastatic lesions of animals . Collectively , we provide evidence of isolation , characterization , and long-term culture of human breast cancer CTCs , leading to the description of a BMSM protein signature that is suggestive of CTC metastatic competency to the brain . Q96EB6 activation enhances HDAC inhibition-mediated upregulation of O95257 by repressing the binding of NF-κB/ P40763 complex to its promoter in malignant lymphoid cells . We explored the activity of Q96EB6 activators ( DB05073 and SRT2183 ) alone and in combination with DB06603 in a panel of malignant lymphoid cell lines in terms of biological and gene expression responses . DB05073 and SRT2183 induced growth arrest and apoptosis , concomitant with deacetylation of P40763 and NF-κB , and reduction of c-Myc protein levels . PCR arrays revealed that SRT2183 leads to increased mRNA levels of pro-apoptosis and DNA-damage-response genes , accompanied by accumulation of phospho-H2A.X levels . Next , ChIP assays revealed that SRT2183 reduces the DNA-binding activity of both NF-κB and P40763 to the promoter of O95257 , which is one of the most upregulated genes following SRT2183 treatment . Combination of SRT2183 with DB06603 enhanced the anti-growth and anti-survival effects mediated by either compound alone . Quantitative-PCR confirmed that the DB06603 in combination with SRT2183 , DB05073 or resveratrol leads to greater upregulation of O95257 than any of the single agents . DB06603 plus SRT2183 in combination showed greater inhibition of c-Myc protein levels and phosphorylation of H2A.X , and increased acetylation of p53 . Furthermore , EMSA revealed that NF-κB binds directly to the O95257 promoter , while P40763 binds indirectly in complexes with NF-κB . In addition , the binding of NF-κB/ P40763 complexes to the O95257 promoter is inhibited following DB06603 , DB05073 or resveratrol treatment . Moreover , the combination of DB06603 with SRT2183 , DB05073 or resveratrol induces a greater binding repression than either agent alone . These data suggest that P40763 is a corepressor with NF-κB of the O95257 gene and provides in vitro proof-of-concept for the combination of HDACi with Q96EB6 activators as a potential new therapeutic strategy in lymphoid malignancies . Inhibition of human steroid 5beta-reductase ( P51857 ) by finasteride and structure of the enzyme-inhibitor complex . The Delta(4)-3-ketosteroid functionality is present in nearly all steroid hormones apart from estrogens . The first step in functionalization of the A-ring is mediated in humans by steroid 5alpha- or 5beta-reductase . DB01216 is a mechanism-based inactivator of 5alpha-reductase type 2 with subnanomolar affinity and is widely used as a therapeutic for the treatment of benign prostatic hyperplasia . It is also used for androgen deprivation in hormone-dependent prostate carcinoma , and it has been examined as a chemopreventive agent in prostate cancer . The effect of finasteride on steroid 5beta-reductase ( P51857 ) has not been previously reported . We show that finasteride competitively inhibits P51857 with low micromolar affinity but does not act as a mechanism-based inactivator . The structure of the P51857 .NADP(+)*finasteride complex determined at 1.7 A resolution shows that it is not possible for NADPH to reduce the Delta(1-2)-ene of finasteride because the cofactor and steroid are not proximal to each other . The P01024 -ketone of finasteride accepts hydrogen bonds from the catalytic residues DB00135 -58 and DB00142 -120 in the active site of P51857 , providing an explanation for the competitive inhibition observed . This is the first reported structure of finasteride bound to an enzyme involved in steroid hormone metabolism . DB06589 inhibits the activation of P09619 β-expressing astrocytes in the brain metastatic microenvironment of breast cancer cells . Brain metastases occur in more than one-third of metastatic breast cancer patients whose tumors overexpress P04626 or are triple negative . Brain colonization of cancer cells occurs in a unique environment , containing microglia , oligodendrocytes , astrocytes , and neurons . Although a neuroinflammatory response has been documented in brain metastasis , its contribution to cancer progression and therapy remains poorly understood . Using an experimental brain metastasis model , we characterized the brain metastatic microenvironment of brain tropic , P04626 -transfected MDA-MB-231 human breast carcinoma cells ( 231-BR- P04626 ) . A previously unidentified subpopulation of metastasis-associated astrocytes expressing phosphorylated platelet-derived growth factor receptor β ( at tyrosine 751 ; p751- P09619 β ) was identified around perivascular brain micrometastases . p751- P09619 β(+) astrocytes were also identified in human brain metastases from eight craniotomy specimens and in primary cultures of astrocyte-enriched glial cells . Previously , we reported that pazopanib , a multispecific tyrosine kinase inhibitor , prevented the outgrowth of 231-BR- P04626 large brain metastases by 73 % . Here , we evaluated the effect of pazopanib on the brain neuroinflammatory microenvironment . DB06589 treatment resulted in 70 % ( P = 0.023 ) decrease of the p751- P09619 β(+) astrocyte population , at the lowest dose of 30 mg/kg , twice daily . Collectively , the data identify a subpopulation of activated astrocytes in the subclinical perivascular stage of brain metastases and show that they are inhibitable by pazopanib , suggesting its potential to prevent the development of brain micrometastases in breast cancer patients . In vivo magnetomotive optical molecular imaging using targeted magnetic nanoprobes . Dynamic magnetomotion of magnetic nanoparticles ( MNPs ) detected with magnetomotive optical coherence tomography ( MM- O75051 ) represents a new methodology for contrast enhancement and therapeutic interventions in molecular imaging . In this study , we demonstrate in vivo imaging of dynamic functionalized iron oxide MNPs using MM- O75051 in a preclinical mammary tumor model . Using targeted MNPs , in vivo MM- O75051 images exhibit strong magnetomotive signals in mammary tumor , and no significant signals were measured from tumors of rats injected with nontargeted MNPs or saline . The results of in vivo MM- O75051 are validated by Q9BWK5 , ex vivo MM- O75051 , DB06783 staining of histological sections , and immunohistochemical analysis of excised tumors and internal organs . The MNPs are antibody functionalized to target the human epidermal growth factor receptor 2 ( P04626 neu ) protein . Fc-directed conjugation of the antibody to the MNPs aids in reducing uptake by macrophages in the reticulo-endothelial system , thereby increasing the circulation time in the blood . These engineered magnetic nanoprobes have multifunctional capabilities enabling them to be used as dynamic contrast agents in MM- O75051 and Q9BWK5 . Retreatment of men with metastatic castrate-resistant prostate cancer with abiraterone . BACKGROUND : DB05812 acetate ( AA ) , oral P05093 inhibitor , is an active agent in the treatment of metastatic castrate-resistant prostate cancer ( mCRPC ) . METHODS : We ( R.L.A and N.A ) retrospectively evaluated outcome in 12 men who were re-treated with AA following prior treatment with AA at the Princess Margaret Cancer Centre . RESULTS : All men were heavily pre-treated for mCRPC with a median of four prior lines of therapy , one of which was AA ( given either pre- or post-chemotherapy ) . Eleven out of 12 ( 92 % ) men stopped their first treatment course of AA due to progression and one stopped for financial reasons . Seven men had a PSA decrease ≥50 % following their first AA treatment , of which three ( 46 % ) had a PSA decrease ≥50 % to AA re-treatment . The responses to AA re-treatment were generally short-lived with a median biochemical progression-free survival of 2.3 months and median treatment duration of 3.2 months . No PSA responses to AA re-treatment were seen in five men who did not have an initial PSA response to AA . CONCLUSIONS : Our data suggest that AA re-challenge may have limited benefit in select men with mCRPC , and warrants further formal research . Genetic polymorphisms of P03372 , Q92731 , P05093 , and P11511 and the risk of breast cancer : a case control study from North India . Estrogen is a key driver of breast cancer and genes involved in its signaling and biosynthesis are crucial in breast cancer progression . In this study , we investigated the role of estrogen signaling and synthesis related genes polymorphism in susceptibility to breast cancer risk in North India population in a case-control approach . We examined the association of single nucleotide polymorphism ( SNP ) in estrogen receptors , P03372 ( rs2234693 ) and Q92731 ( rs2987983 ) ; estrogen biosynthesis enzymes , P05093 ( rs743572 ) ; and aromatase , P11511 ( rs700519 ) with breast cancer risk . Cases ( n = 360 ) were matched to controls ( n = 360 ) by age , sex , ethnicity , and geographical location . Results provided evidence that all the genetic variants were significantly associated with breast cancer risk among North Indian women . Furthermore , on performing stratified analysis between breast cancer risk and different clinicopathological characteristics , we observed strong associations for menopausal status , estrogen receptor ( ER ) , progesterone receptor ( PR ) , human epidermal growth factor receptor 2 ( P04626 ) status , clinical stage , and histological grade . Our results suggest that these genes could be used as molecular markers to assess breast cancer susceptibility and predicting prognosis in North India population . DB00183 infusions in patients with panic disorder . II. Neuroendocrinology . Cholecystokinin ( CCK ) has well-documented anxiogenic effects in animals and normal people , and panicogenic effects in patients with panic disorder , but little is known about its neuroendocrine profile . We examined neuroendocrine responses to intravenous infusions of pentagastrin , a selective P32239 agonist , in 10 patients with panic disorder and 10 normal control subjects . DB00183 potently activated the hypothalamic-pituitary-adrenal ( Q9Y251 ) axis , but did not release growth hormone or any of several vasoactive peptides ( neurokinin A , DB05875 , vasoactive intestinal peptide ) . The Q9Y251 axis response was unrelated to increases in symptoms . Panic patients did not differ from controls in neuroendocrine responses to the CCK agonist . Differential sensitivity to novelty stress accounted for the only patient-control differences in neuroendocrine profiles . The data suggest that CCK may help modulate normal Q9Y251 axis activity , but its anxiogenic effects are unrelated to its stimulatory effects on the Q9Y251 axis . DB00183 provides a safe and readily available probe for further study of CCK receptor systems in humans . Synergistic proapoptotic effects of the two tyrosine kinase inhibitors pazopanib and lapatinib on multiple carcinoma cell lines . DB06589 and lapatinib are two tyrosine kinase inhibitors that have been designed to inhibit the P15692 tyrosine kinase receptors 1 , 2 and 3 ( pazopanib ) , and the P00533 and P04626 receptors in a dual manner ( lapatinib ) . DB06589 has also been reported to mediate inhibitory effect on a selected panel of additional tyrosine kinases such as P09619 and c-kit . Here , we report that pazopanib and lapatinib act synergistically to induce apoptosis of A549 non-small-cell lung cancer cells . Systematic assessment of the kinome revealed that both pazopanib and lapatinib inhibited dozens of different tyrosine kinases and that their combination could suppress the activity of some tyrosine kinases ( such as c- DB00134 ) that were not or only partially affected by either of the two agents alone . We also found that pazopanib and lapatinib induced selective changes in the transcriptome of A549 cells , some of which were specific for the combination of both agents . Analysis of a panel of unrelated human carcinoma cell lines revealed a signature of 52 genes whose up- or downregulation reflected the combined action of pazopanib and lapatinib . Indeed , pazopanib and lapatinib exerted synergistic cytotoxic effects on several distinct non-small-cell lung cancer cells as well as on unrelated carcinomas . Altogether , these results support the contention that combinations of tyrosine kinase inhibitors should be evaluated for synergistic antitumor effects . Such combinations may lead to a ' collapse ' of pro-survival signal transduction pathways that leads to apoptotic cell death . Curcumin and epigallocatechin gallate inhibit the cancer stem cell phenotype via down-regulation of P40763 -NFκB signaling . BACKGROUND/AIM : The cancer stem cell ( CSC ) model postulates the existence of a small proportion of cancer cells capable of sustaining tumor formation , self-renewal and differentiation . Signal Transducer and Activator of Transcription 3 ( P40763 ) signaling is known to be selectively activated in breast CSC populations . However , it is yet to be determined which molecular mechanisms regulate P40763 signaling in CSCs and what chemopreventive agents are effective for suppressing CSC growth . The aim of this study was to examine the potential efficacy of curcumin and epigallocatechin gallate ( EGCG ) against CSC and to uncover the molecular mechanisms of their anticancer effects . MATERIALS AND METHODS : To suppress the CSC phenotype , two breast cancer cell lines ( MDA-MB-231 cells and MCF7 cells transfected with P04626 ) were treated with curcumin ( 10 μM ) with or without EGCG ( 10 μM ) for 48 h . We used tumor-sphere formation and wound-healing assays to determine CSC phenotype . To quantify CSC populations , Fluorescence-activated cell sorting profiling was monitored . P40763 phosphorylation and interaction with Nuclear Factor-kB ( NFkB ) were analyzed by performing western blot and immunoprecipitation assays . RESULTS : Combined curcumin and EGCG treatment reduced the cancer stem-like Cluster of differentiation 44 ( P16070 ) -positive cell population . Western blot and immunoprecipitation analyses revealed that curcumin and EGCG specifically inhibited P40763 phosphorylation and P40763 -NFkB interaction was retained . CONCLUSION : This study suggests that curcumin and EGCG function as antitumor agents for suppressing breast CSCs . P40763 and NFκB signaling pathways could serve as targets for reducing CSCs leading to novel targeted-therapy for treating breast cancer . DB06612 and eosinophil-mediated disease . Eosinophils are major pro-inflammatory cells that make a major contribution to diseases that affect the upper and lower airways , skin and gastrointestinal tract . Interleukin ( IL ) -5 is central to their maturation and release from the bone marrow together with their subsequent accumulation and activation in the tissues . DB06612 is a humanized monoclonal antibody ( mAb ) with potent P05113 neutralizing effects that represents a potential treatment for eosinophilic diseases . Several clinical trials with mepolizumab reported that treatment of patients with mild to severe asthma resulted in a substantial reduction in blood and sputum eosinophil numbers . However , clinical outcomes were disappointing as there were no significant effects on airway hyper-reactivity or the late asthmatic reaction to inhaled allergen challenge . More recently two studies , one in in patients with refractory eosinophilic asthma with a history of recurrent severe exacerbations and the other in patients with persistent sputum eosinophilia and symptoms despite systemic treatment with prednisone treatment , reported that monthly intravenous mepolizumab reduced sputum/blood eosinophilia , asthma exacerbations together with improvments in quality of life . DB06612 also appears to be an effective therapy for hypereosinophilic syndrome while other trials have shown efficacy of mepolizumab therapy in eosinophilic esophagitis . This review will consider the current status of the clinical development of mepolizumab for diseases with a significant eosinophilic component to their pathology . Characterization of proapoptotic compounds from the bark of Garcinia oblongifolia . Twenty compounds from Garcinia oblongifolia were screened for proapoptotic activity using FRET-based HeLa- P01024 sensor cells . Among them , oblongifolins F and G ( 1 and 2 ) , 1,3,5-trihydroxy-13,13-dimethyl-2H-pyran[7,6-b]xanthone ( 3 ) , nigrolineaxanthone T ( 4 ) , and garcicowin B ( 5 ) showed significant proapoptotic activity at a concentration of 10 μM . Bioassessments were then performed to evaluate the potential of these compounds for therapeutic application . All five compounds showed significant cytotoxicity and caspase-3-activating ability in cervical cancer HeLa cells , with compounds 1 and 2 having the highest potencies . All five compounds specifically induced caspase-dependent apoptosis , which could be prevented by the pan-caspase inhibitor zVAD-fmk . In particular , 3 induced apoptosis through mitotic arrest . Compounds 1-5 displayed similar IC50 values ( 3.9-16.5 μM ) against the three cancer cell lines HeLa , MDA-MB-435 , and HepG2 . In addition , compounds 1 , 2 , and 4 exhibited similar and potent IC50 values ( 2.4-5.1 μM ) against several breast and colon cancer cell lines , including those overexpressing either P04626 or P-glycoprotein . P04626 and P-glycoprotein are known factors that confer resistance to anticancer drugs in cancer cells . This is the first study on the cytotoxicity , caspase-3-activing ability , and specificity of proapoptotic compounds isolated from G. oblongifolia in HeLa cells . The potential application of these compounds against P04626 - or P-glycoprotein-overexpressing cancer cells was investigated . Recent advances in the regulation of matrix metalloproteinase 2 activation : from basic research to clinical implication ( Review ) . Matrix metalloproteinases ( MMPs ) play an important role in degradation of extracellular matrix ( Q13201 ) , which is an essential step in the cascade of metastasis . Various types of MMPs are expressed and activated in head and neck squamous cell carcinoma ( HNSCC ) as well as other human cancers . P08253 is a prominent predictor of poor prognosis . Membrane type 1-MMP ( P50281 ) was originally identified as an activator of P08253 . In addition to the original role , recent studies show other important functions of P50281 such as degradation of type I collagen and cleavage of P16070 . Tissue inhibitor of P08253 ( P16035 ) was identified as an inhibitor of P08253 and P50281 . However , P16035 was reported to be essential for cell-mediated activation of P08253 , and thus the contribution of P16035 to tumor invasion has remained controversial . Some studies also suggested a role of P16035 as a predictor of poor prognosis . Thus , inhibition of MMP activation by TIMPs is not a suitable strategy for suppressing invasion and metastasis . Instead of P16035 , various MMP inhibitors ( MMPI ) such as DB00786 have been investigated with regard to suppression of tumor progression and improvement of prognosis in patients with advanced cancers , which resulted in no clinical efficacy . MMPs are especially important in the early stage of cancer progression , and thus strategies for future MMPI trials should be reconsidered .

MH_train_1366

MH_train_1366

MH_train_1366

interacts_with DB08816?

multiple_choice

Role of costimulatory molecules in immune response of patients with cutaneous leishmaniasis . T cell-mediated immunity is critical in resistance against Leishmania parasites , and T cell activation requires signals provided by costimulatory molecules . Herein we evaluated the role of costimulatory molecules on cytokine production and T cell surface molecule expression by peripheral blood mononuclear cells ( PBMC ) from cutaneous leishmaniasis ( CL ) patients . PBMC from CL patients were stimulated with soluble Leishmania antigen ( SLA , 10 microg/ml ) , in the presence or absence of soluble P16410 -Ig to block P10747 - P33681 interaction or in the presence or absence of anti-human P29965 to block P25942 - P29965 interaction . Supernatants were harvested to evaluate tumor necrosis factor alpha ( P01375 ) , interleukin 10 ( P22301 ) , transforming growth factor beta ( TGF-beta ) and interferon gamma ( P01579 ) production by ELISA . Cells were harvested after 48 h of culture , stained for specific activation markers and analyzed by flow cytometry . Results show that the blockade of P10747 - P33681 interaction by P16410 -Ig downmodulated P01579 , P22301 , and P01375 secretion by PBMC from CL patients . No alteration was detected on either TGF-beta production or the expression of CTLA44 or CD25 on P01730 + and CD8+ T cells . When the P25942 - P29965 interaction was blockade using anti- P29965 , we did not observe changes in cytokine production or in surface molecule expression . The blockade of the P10747 - P33681 interactions by P16410 -Ig also did not alter cytokine production in volunteers immunized against tetanus toxoid ( TT ) . Taken together , these data suggest that the interaction of P16410 and P10747 - P33681 is a TGF-beta-independent mechanism that specifically downmodulates the immune response in cutaneous leishmaniasis patients . EBV infection induces expression of the transcription factors P39905 -2/c-Jun in B lymphocytes but not in B-CLL cells . B cell type chronic lymphocytic leukaemia ( B-CLL ) cells carry the Epstein-Barr virus ( EBV ) receptor CD21 and can be infected in vitro with the virus . The infected cells exhibit an unusual EBV program , they express the nuclear proteins but not latent membrane protein 1 ( Q9NR12 -1 ) . Similar cells were encountered in lymphoid tissues of infectious mononucleosis ( IM ) patients and in lymphoproliferations of immunosuppressed patients . EBV infected B-CLL cells can be regarded as model for this viral program . In B cells the regulation of Q9NR12 -1 is executed mainly by EBV encoded nuclear antigen 2 ( EBNA-2 ) , interacting with several cellular proteins and these complexes bind to specific sequences in the Q9NR12 -1 promoter . P15336 and c-Jun were shown to be among the interacting partners of EBNA-2 . These molecules can be detected in experimentally infected B lymphocytes . We found c-Jun and/or phosphorylated P39905 -2 ( p- P39905 -2 ) expression in some B-CLL ex vivo samples . They disappeared or their expression declined promptly in explanted cells , even if they were infected with EBV in vitro . Activation of the infected B-CLL cells by exposure to P29965 was accompanied by p- P39905 -2 and c-Jun but not by Q9NR12 -1 expression . In one of three clones tested , subsequent treatment with histone deacetylase inhibitors ( HDACi ) , P32119 or n-butyrate , could induce Q9NR12 -1 . Treatment with phorbol-12 , 13-dibutyrate ( PDB ) induced Q9NR12 -1 expression in three of four clones . Neither the HDACi nor the PDB treated cells survived . P30968 and peritoneal plasmin activity . Most surgical procedures performed by obstetrician-gynecologists are associated with pelvic adhesions that cause subsequent serious sequelae , including small bowel obstruction , infertility , chronic pelvic pain , and difficulty in postoperative treatment , including complexity during subsequent surgical procedures . This study was conducted to determine if gonadotropin-releasing hormone analogues ( GnRHa ) affect the expressing tissue-type plasminogen activator ( t-PA ) and its inhibitor-1 ( P05121 ) in peritoneal cells in culture . Human peritoneal Met5A cells were used to examine the effects of GnRHa leuprolide , buserelin and goserelin on the levels of t-PA and PA-1 . Antigen concentrations were measured in conditioned media and cell lysates by real-time PCR and ELISA . P30968 ( GnRHR ) mRNA was determined by RT-PCR . GnRHR mRNA was detected in Met5A cells . Exposure of Met5A cells to GnRHa induced a rapid decrease of P05121 level in cultured medium but not in cell lysate ( protein and mRNA ) . These effects of GnRHa on P05121 were not associated with any changes in t-PA level . These results suggest that GnRHa may be an effective stimulator of local peritoneal fibrinolytic activity , as it decreases P05121 secretion in peritoneal Met5A cells by a mechanism linked to GnRHR . Evaluation of degarelix in the management of prostate cancer . Medical castration using gonadotropin-releasing hormone ( DB00644 ) receptor agonists currently provides the mainstay of androgen deprivation therapy for prostate cancer . Although effective , these agents only reduce testosterone levels after a delay of 14 to 21 days ; they also cause an initial surge in testosterone that can stimulate the cancer and lead to exacerbation of symptoms ( " clinical flare " ) in patients with advanced disease . Phase III trial data for the recently approved P30968 blocker , degarelix , demonstrated that it is as effective and well tolerated as DB00644 agonists . However , it has a pharmacological profile more closely matching orchiectomy , with an immediate onset of action and faster testosterone and PSA suppression , without a testosterone surge or microsurges following repeated injections . As a consequence , with this DB00644 blocker , there is no risk of clinical flare and no need for concomitant antiandrogen flare protection . DB06699 therefore provides a useful addition to the hormonal armamentarium for prostate cancer and offers a valuable new treatment option for patients with hormone-sensitive advanced disease . Here , we review key preclinical and clinical data for degarelix , and look at patient-focused perspectives in the management of prostate cancer . P11511 inhibitors and cyclooxygenase-2 ( P35354 ) inhibitors in endometriosis : new questions -- old answers ? The medical treatment of endometriosis needs to be optimized . Therapeutic management strategies for endometriosis-associated pain or recurrent disease are primarily aimed at downregulating ovarian function or antagonizing the effect of estrogen in ectopic endometrial implants . In this context , basic research is providing important results for the development of new , specific treatment modalities . P11511 overexpression has recently been detected in endometriotic tissue . P11511 ( p450arom ) is responsible for converting C19 androgens into estrogen in several types of human tissue . P11511 activity causes local estrogen biosynthesis , which , in turn , stimulates prostaglandin E2 production by upregulating cyclooxygenase-2 ( P35354 ) . Thus , a positive feedback cycle develops between the two systems . Another abnormality in endometriosis , the deficient 17beta-hydroxysteroiddehydrogenase type II ( 17beta-HSD-Type-II ) expression , impairs the inactivation of estradiol to estrone . In contrast to the eutopic endometrium , these molecular aberrations increase the amount of local estradiol and prostaglandin E2 in endometriosis . In several human cell lines , prostaglandin and estrogen concentrations are associated with proliferation , migration , angiogenesis , apoptosis resistance and even invasiveness . Consequently , aromatase and P35354 are thought to be promising new therapeutic targets . Thus , specific aromatase inhibitors ( e.g. DB01006 / DB01006 , DB01217 /Arimidex or Exemestan/Aromasin ) or selective P35354 inhibitors ( e.g. Celecoxib/ DB00482 , DB00533 /Vioxx , DB00580 /Bextra ) are of great interest and should be studied in clinical trials in premenopausal woman with endometriosis to expand the spectrum of currently available treatment options . Glycoprotein IIb/IIIa and Q9H244 receptor antagonists yield additive inhibition of platelet aggregation , granule secretion , soluble P29965 release and procoagulant responses . Glycoprotein IIb/IIIa ( P08514 /IIIa ) antagonists , including abciximab and tirofiban , are administered concurrently with clopidogrel , a Q9H244 antagonist , and aspirin in some patients undergoing percutaneous coronary intervention . We studied the effects of , and interactions between , abciximab , tirofiban , aspirin and the Q9H244 antagonist cangrelor on platelet aggregation , alpha and dense granule secretion and procoagulant responses in vitro . Blood was obtained from healthy volunteers . Platelet aggregation , dense granule secretion , alpha granule secretion ( P05121 and soluble P29965 levels ) and procoagulant responses ( annexin-V and microparticle formation ) were assessed using collagen and thrombin receptor activating peptide ( TRAP ) as agonists . All the antagonists used singularly inhibited collagen-induced responses . Combinations of abciximab or tirofiban with aspirin and/or cangrelor gave additive inhibition with the greatest effect seen when abciximab or tirofiban was combined with both aspirin and cangrelor . DB06441 inhibited TRAP-induced responses and , again , there was additive inhibition of these parameters when abciximab or tirofiban were combined with cangrelor . The P08514 /IIIa receptor plays an important role in amplification of platelet activation such that there are important interactions between P08514 /IIIa antagonists and inhibitors of both Q9H244 receptor activation and , to a lesser extent , thromboxane A2 generation . These interactions are likely to have important influences on the safety and efficacy of combination anti-platelet therapies . DB00173 nucleotides inhibit cytokine generation by human mast cells through a Gs-coupled receptor . DB00171 and ADP activate functionally distinct G protein-coupled purinergic ( P2Y ) receptors . We determined the expression and function of adenine nucleotide-specific P2Y receptors on cord blood-derived human mast cells ( hMCs ) . Human MCs expressed mRNA encoding the ADP-specific P47900 , Q9H244 , and Q9BPV8 receptors ; the DB00171 /UTP-specific P41231 receptor ; and the DB00171 -selective Q96G91 receptor . ADP ( 0.05-50 muM ) induced calcium flux that was completely blocked by a P47900 receptor-selective antagonist and was not cross-desensitized by DB00171 . Low doses of ADP induced strong phosphorylation of P29323 and p38 MAPKs ; higher doses stimulated eicosanoid production and exocytosis . Although MAPK phosphorylation was blocked by a combination of P47900 - and Q9H244 -selective antagonists , neither interfered with secretion responses . Unexpectedly , both ADP and DB00171 inhibited the generation of P01375 in response to the O60603 ligand , peptidoglycan , and blocked the production of P01375 , P10145 , and MIP-1beta in response to leukotriene D(4) . These effects were mimicked by two DB00171 analogues , adenosine 5'-O-(3-thiotriphosphate) and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate ( BzATP ) , but not by adenosine . ADP , DB00171 , adenosine 5'-O-(3-thiotriphosphate) , and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate each induced DB02527 accumulation , stimulated the phosphorylation of CREB , and up-regulated the expression of inducible DB02527 early repressor , a CREB-dependent inhibitor of cytokine transcription . Human MCs thus express several ADP-selective P2Y receptors and at least one G(s)-coupled ADP/ DB00171 receptor . Nucleotides could therefore contribute to MC-dependent microvascular leakage in atherosclerosis , tissue injury , and innate immunity while simultaneously limiting the extent of subsequent inflammation by attenuating the generation of inducible cytokines by MCs . Q9Y2D1 polymorphisms influence P39905 function and response to treatment in children with childhood acute lymphoblastic leukemia . DB00023 is a standard and critical component in the therapy of childhood acute lymphoblastic leukemia . DB00174 synthetase ( P08243 ) and the basic region leucine zipper activating transcription factor 5 ( Q9Y2D1 ) and arginosuccinate synthase 1 ( P00966 ) have been shown to mediate the antileukemic effect of asparaginase and to display variable expression between leukemia cells that are resistant and sensitive to treatment . Fourteen polymorphisms in the regulatory and coding regions of these genes were investigated for an association with acute lymphoblastic leukemia outcome . Lower event-free survival ( O43281 ) was associated with Q9Y2D1 T1562C , tandem-repeat P08243 polymorphism , derived haplotype , and P00966 G1343T and G34T substitutions ( P ≤ .03 ) . Associations were limited to patients who received Escherichia coli asparaginase . Variations that sustained correction for multiple testing ( Q9Y2D1 T1562C , P = .005 ; P08243 tandem-repeat and related haplotype , P ≤ .01 ) were subsequently analyzed in the replication cohort . The E coli-dependent association of the Q9Y2D1 T1562 allele with reduced O43281 was confirmed ( P = .01 ) . A gene-reporter assay showed that the haplotype tagged by T1562 had higher promoter activity ( P ≤ .01 ) . The remaining regulatory polymorphisms also appeared to affect Q9Y2D1 function ; 2 additional high-activity haplotypes were identified ( P ≤ .02 ) and were further corroborated by quantitative mRNA analysis in lymphoblastoid cell lines . The Q9Y2D1 -regulated increase in P08243 expression in response to more efficacious E coli-induced asparagine depletion may explain our observed results . [ Prominent features of management strategies in acute coronary syndromes with the new oral antiplatelet agents ] . The novel oral Q9H244 inhibitors ( prasugrel and ticagrelor ) have been incorporated into the recently updated acute coronary syndrome ( ACS ) guidelines , as an adjunct antiplatelet treatment to aspirin . The studies involving the use of new oral antiplatelet agents that are more potent , predictable and faster platelet inhibitors than clopidogrel have demonstrated superiority with respect to the primary composite endpoint ( cardiovascular death , non-lethal myocardial infarction , stroke ) for both prasugrel and ticagrelor compared to clopidogrel . The subgroup analysis of the relevant studies showed that these new agents differ in their level of efficacy in different ACS patient subgroups : ( 1 ) Mortality was reduced with ticagrelor ; ( 2 ) DB08816 is especially more effective in intermediate-and high-risk non-ST elevation ACS patients in whom early invasive strategy is selected ; ( 3 ) Prasugrel should be especially preferred in patients with acute ST elevation myocardial infarction undergoing percutaneous coronary intervention ( P05154 ) after diagnostic angiography ; and ( 4 ) Prasugrel is more effective in diabetic patients . While clopidogrel is recommended for ACS patients who are followed with a non-invasive strategy or who have not undergone percutaneous revascularization , it is the last line choice or an alternative to the Q9H244 inhibitor therapy for patients undergoing invasive strategy . Direct oral anticoagulants in acute coronary syndrome . Patients with acute coronary syndromes ( ACS ) require a specific antithrombotic therapy in the immediate and the post ACS phase . The current antithrombotic therapy in the acute phase of an ACS combines antiplatelet and anticoagulant drugs in order to reduce ischemic cardiovascular events . In the post ACS phase , dual antiplatelet therapy ( DAPT ; aspirin and a Q9H244 receptor antagonist ) is the current mainstay of antithrombotic treatment and is recommended in the guidelines of the major North American and European clinical cardiology associations ( DB00551 , ACC , and ESC ) . Recently , the addition of rivaroxaban , a low dose oral direct factor Xa inhibitor ( 2.5 mg twice daily ) , to DAPT ( aspirin plus second-generation Q9H244 inhibitor ) showed a significant reduction of cardiovascular and overall mortality in the major phase III clinical trial ATLAS ACS 2 TIMI 51 . This led to the approval of low-dose rivaroxaban in addition to aspirin and clopidogrel by the European Medicines Agency ( P15941 ) in 2013 . Other direct oral anticoagulants ( apixaban , dabigatran etexilate ) have also been assessed in phase II ( dabigatran etexilate ) and phase III ( apixaban ) post ACS clinical trials . In the studied dosing regimens , these drugs failed to show a net clinical benefit in addition to dual antiplatelet therapy . The major clinical phase II and III post ACS studies of direct oral anticoagulants are summarized and discussed in this article along with the concept of long-term anticoagulation for the secondary prevention of ischemic events after ACS and implications for the future of antithrombotic therapy in the current era of third-generation Q9H244 receptor inhibitors ( Prasugrel and DB08816 ) . A field synopsis and meta-analysis of genetic association studies in peripheral arterial disease : The CUMAGAS-PAD database . In an electronic search of the literature , the authors systematically retrieved all published studies that investigated genetic susceptibility to peripheral arterial disease ( PAD ) . They created a comprehensive database of all eligible studies , collecting detailed genetic and bioinformatics data on each polymorphism . Data from eligible studies were synthesized using meta-analysis techniques . Gene variants were classified into distinct pathophysiologic pathways , and their potential involvement in PAD pathogenesis was determined . Forty-one publications that examined 44 gene polymorphisms were included . For 37 polymorphisms , the variant form had a functional effect . Twenty-three polymorphisms in 22 potential PAD candidate genes ( F2 , P02675 , P42898 , P05106 , P12821 , AGT , P05231 , P13500 , P05362 , P16581 , P14780 , P37231 , P03956 , P35611 , Q9H244 , P11150 , Q13093 , Q8WTV0 , P08254 , P55157 , P08519 , P32297 ) showed a significant association in individual studies . Eighty-eight percent of the studies had statistical power of less than 50 % , and in 15 studies the genotype distribution in the control group did not conform to Hardy-Weinberg equilibrium . Data on 12 polymorphisms ( P12259 1691 G/A , P42898 677C/T , F2 20210 G/A , P05106 1565 T/C , P12821 I/D , AGT 704C/T , AGT -6G/A , AGT 733C/T , P05231 -174 G/C , P14780 -1562C/T , P05362 1462A/G , P32297 831C/T ) were synthesized , and a positive association was found for 3 ( P05231 -174 G/C , P05362 1462A/G , P32297 831C/T ) . Administration of adenosine diphosphate-ribosyl transferase antagonist allows in vivo control of anti-dinitrophenyl response . DB03073 ( 3MB ) is one of a series of chemical inhibitors of the nuclear enzyme adenosine diphosphate ( ADP ) -ribosyl transferase ( P09874 ) , which has been shown to inhibit cell differentiation in vitro , but has no effect on differentiation independent proliferation . Treatment of mice with an optimal concentration of 3MB ( 20 mg/kg body weight ) at or 1 day after dinitrophenyl-keyhole limpet haemocyanin ( DNP-KLH ) immunisation reduced anti-DNP plaque-forming cell ( P27918 ) numbers to less than 10 % of those of control animals . The period for maximum P27918 suppression showed a narrow time window relative to immunisation , suggesting that in vivo , as in vitro , 3 MB was acting only on those lymphocytes differentiating in response to antigen . Experimental findings showed that it was possible to select for P27918 derived from different populations of DNP-responsive lymphocytes by adjusting the time of 3MB treatment relative to immunisation . When 3MB was used with antigen priming , the residual P27918 showed a lower average affinity than P27918 in mice treated with 3MB 3 days after priming , suggesting a differential selection of those lymphocytes responding either ' early ' or ' late ' in the primary immune response . The expression of the mitochondrial gene P03905 is downregulated in cystic fibrosis . Cystic fibrosis ( CF ) is a disease produced by mutations in the P13569 channel . We have previously reported that the P13569 chloride transport activity indirectly regulates the differential expression of several genes , including P12931 and P15941 . Here we report that P03905 , a mitochondrial gene encoding a subunit of the mitochondrial Complex I ( mtCx-I ) , is also a P13569 -dependent gene . A reduced expression of P03905 was observed in CFDE cells ( derived from a CF patient ) when compared to CFDE cells ectopically expressing wild-type P13569 . The differential expression of P03905 in CF was confirmed by RT-PCR . In situ hybridizations of deparaffinized human lung tissue slices derived from wt- P13569 or CF patients also showed downregulation of P03905 in CF . In addition , the P13569 chloride transport inhibitors glibenclamide and P13569 (inh)-172 also reduced P03905 expression in CFDE cells ectopically expressing wt P13569 . These results suggest that the P13569 chloride transport activity indirectly up-regulates P03905 expression . DB04690 inhibits Tat-mediated transactivation of type 1 human immunodeficiency virus . Transcription of type 1 human immunodeficiency virus ( HIV-1 ) is governed by the viral long terminal repeat ( LTR ) . By using HIV-1 LTR-directed reporter gene systems , we found that the P11387 inhibitor camptothecin inhibits Tat-mediated transactivation of HIV-1 LTR . The 293.27.2 cells that carry a stably transfected HIV-1 LTR-directed lacZ gene expression vector ( pNAZ ) were used . Inhibitions of LTR were observed at camptothecin concentrations ( IC50 about 0.03 microM , which was an order of magnitude lower than for Ro 24-7429 ) , which had minor effects on cell survival , expression of the cellular gene gro , or Rous sarcoma virus-directed chloramphenicol acetyltransferase ( CAT ) gene expression . Inhibition was also seen with RPMI 8402 , which is a human P01730 -positive lymphocyte line transiently transfected with a HIV-1 LTR-directed ( CAT ) gene . Experiments with HIV-1 LTR mutants suggest that transactivation response sequence but not NF-kappa B is responsible for the inhibition by camptothecin . The target for camptothecin may be a cellular factor that is important for the activation of HIV-1 LTR by Tat and thus may offer a potential target for therapy of HIV-1 infection . Effects of LSD on Ca++ currents in central 5-HT-containing neurons : P08908 receptors may play a role in hallucinogenesis . Drugs that influence the activity of central serotonergic neurons by activating a 5-hydroxytryptamine subtype of receptor ( P08908 ) alter mood and perception . Previously , we demonstrated with whole-cell recordings from acutely isolated 5-HT-containing dorsal raphe ( DR ) neurons from the adult rat that 5-HT inhibited Ca++ current and activated K+ current in DR neurons . We now show that DB04829 ( LSD ) mimics the actions of 5-HT ; it dramatically suppresses Ca++ current in a dose-dependent manner and activates an inwardly rectifying K+ conductance . Spiperone ( 0.2 microM ) , a P08908 /5-HT2 antagonist , blocks the effect of both LSD and 5-HT . The nonhallucinogenic structural analog 2-bromo-LSD ( 2-Bol ) at 10 microM has no effect on either Ca++ or K+ current by itself , but it competitively antagonizes both effects of LSD . Inhibition of 5-HT release resulting from P08908 receptor activation may play an integral role in the hallucinogenic actions of LSD by reducing competition between 5-HT and LSD for the postsynaptic 5-HT receptors . DB09036 : first global approval . The anti-interleukin-6 ( P05231 ) chimeric monoclonal antibody siltuximab is the first drug to be approved for the treatment of multicentric Castleman 's disease ( O95822 ) in the US and European union ( EU ) , having gained approval under the FDA priority review program in the US and from an accelerated assessment and recommendation by the Committee for Medicinal Products for Human Use ( CHMP ) in the EU . Development of the drug is continuing in smoldering multiple myeloma . This article summarizes the milestones in the development of siltuximab leading to this first approval for O95822 . Purinergic P47900 receptor signaling mediates wound stimuli-induced cyclooxygenase-2 expression in intestinal subepithelial myofibroblasts . Intestinal subepithelial myofibroblasts ( ISMFs ) are crucial for barrier formation against inflammatory stimuli . Physical injury induces cyclooxygenase-2 ( P35354 ) expression , which accelerates wound healing by ISMFs . However , the mechanism of P35354 induction remains unclear . Physically damaged cells release DB00171 . Here , we investigate the role of DB00171 -purinergic signaling in wound-induced P35354 induction in ISMFs . By 24h post-injury , bovine ISMFs had migrated to and closed the wounded area . A P36551 inhibitor , indomethacin or a purinergic P2 receptor antagonist , suramin , inhibited wound healing . However , additional treatment with indomethacin did not influence wound healing in suramin-treated ISMFs . RT-PCR showed an increase in P35354 mRNA expression 2h post-injury , which was inhibited by suramin . These results suggest that DB00171 mediates wound-induced P35354 elevation . We next assessed the contribution of various purinergic receptors in P35354 induction . An DB00171 analog , ATPγS and a purinergic P47900 , 11-13 receptors agonist , ADP , were among the agents tested which increased P35354 expression . ATPγS-induced P35354 mRNA expression was suppressed by suramin or a purinergic P2Xs , P47900 , 4 , 6 , and 13 receptors antagonist , PPADS . These data suggest the involvement of Gq-coupled purinergic P47900 receptor or Gi-coupled purinergic Q9BPV8 receptor in P35354 induction . U73122 , an inhibitor of phospholipase C , which is a downstream signal of Gq protein , showed suppression of P35354 mRNA expression . However , pertussis toxin , a Gi inhibitor , did not show suppression . We also revealed that inhibitors of p38 MAPK and PKC inhibited ATPγS-induced P35354 mRNA expression . Collectively , purinergic P47900 receptor signaling mediates wound-induced P35354 expression through p38 MAPK and PKC pathways in ISMFs . Analyses of the effects of Gq protein on the activated states of the muscarinic M3 receptor and the purinergic P47900 receptor . G protein-coupled receptors ( GPCRs ) cause various cellular responses through activating heterotrimeric G protein upon the agonist binding . The interaction with G protein has been suggested to stabilize the agonist-bound active conformation of GPCRs . We previously reported the effects of Gq protein on the stabilization of the active conformation of the muscarinic receptor type 1 ( M1R ) , using a fluorescence resonance energy transfer ( FRET ) technique . In this study , we aimed at examining whether or not the binding of Gq protein affects the agonist-induced active conformation of receptors other than the M1R . For this purpose , functionally intact fluorescent receptors of the metabotropic purinergic receptor type 1 ( P2Y1R ) and muscarinic receptor type 3 ( M3R ) were constructed , by inserting junctional linkers between the short intracellular third loops ( i3 ) and yellow fluorescent protein ( YFP ) . The YFP-fused receptors also showed the agonist-induced increases in FRET from the cyan fluorescent protein ( P27918 ) tethered with Gαq subunit , indicating that they interacted with Gq protein . The agonist-induced conformational changes of the receptors were detected as the agonist-induced decrease in FRET between YFP at the i3 and P27918 at the C-tail . The FRET decrease of the M3R but not of the P2Y1R was enhanced by coexpression of Gq protein . In addition , coexpression of Gq protein significantly decelerated the FRET recovery of the M3R construct but not of the P2Y1R construct upon the agonist removal . These results suggest that the effects of the Gq binding on the active conformation of the receptor differ depending on the type of GPCRs . P2Y receptor antagonists in thrombosis . The dual role of P47900 and Q9H244 receptors in platelet aggregation by ADP has been firmly established , based on the action of selective inhibitors , gene targeting in mice and human genetic evidence . Both of these receptor subtypes constitute targets for antithrombotic agents , and compounds with a dual action might also be of interest . However , the agents currently on the market ( ticlopidine and clopidogrel ) , or known to be in development ( cangrelor , DB08816 and prasugrel ) , all target the Q9H244 receptor . The thienopyridines ( ticlopidine , clopidogrel and prasugrel ) irreversibly inactivate the Q9H244 receptor via the covalent binding of an active metabolite generated in the liver , while the other compounds are competitive antagonists . DB06441 , an DB00171 derivative , is suitable for intravenous perfusion , whereas DB08816 is in clinical development as an orally active agent . DB08816 reduces neutrophil recruitment and lung damage in abdominal sepsis . Abstract Platelets play an important role in abdominal sepsis and Q9H244 receptor antagonists have been reported to exert anti-inflammatory effects . Herein , we assessed the impact of platelet inhibition with the Q9H244 receptor antagonist ticagrelor on pulmonary neutrophil recruitment and tissue damage in a model of abdominal sepsis . Wild-type C57BL/6 mice were subjected to cecal ligation and puncture ( CLP ) . Animals were treated with ticagrelor ( 100 mg/kg ) or vehicle prior to CLP induction . Edema formation and bronchoalveolar neutrophils as well as lung damage were quantified . Flow cytometry was used to determine expression of platelet-neutrophil aggregates , neutrophil activation and P29965 expression on platelets . CLP-induced pulmonary infiltration of neutrophils at 24 hours was reduced by 50 % in ticagrelor-treated animals . Moreover , ticagrelor abolished CLP-provoked lung edema and decreased lung damage score by 41 % . Notably , ticagrelor completely inhibited formation of platelet-neutrophil aggregates and markedly reduced thrombocytopenia in CLP animals . In addition , ticagrelor reduced platelet shedding of P29965 in septic mice . Our data indicate that ticagrelor can reduce CLP-induced pulmonary neutrophil recruitment and lung damage suggesting a potential role for platelet antagonists , such as ticagrelor , in the management of patients with abdominal sepsis . Agents with selective estrogen receptor ( ER ) modulator activity induce apoptosis in vitro and in vivo in ER-negative glioma cells . Tamoxifen , a member of the selective estrogen receptor modulator ( SERM ) family , is widely used in the treatment of estrogen receptor ( ER ) -expressing breast cancer . It has previously been shown that high-dose tamoxifen has cytotoxic activity against glioma cells , but whether this effect is drug specific or represents a general property of SERMs is unknown . In this study , we demonstrate that tamoxifen and DB05487 , a novel benzopyranone with SERM activity , induce glioma cell apoptosis in a dose- and time-dependent manner . Moreover , administration of tamoxifen and DB05487 suppresses tumor growth in vivo and extends animal survival in glioma xenograft models . None of the eight glioma cell lines examined express either P03372 or -beta , suggesting the mechanism for tamoxifen- and DB05487 -induced glioma cell apoptosis is independent of the ER signaling pathway . Complementary DNA microarray expression profiling allowed us to identify a subset of genes specifically regulated by tamoxifen and DB05487 , and not by other apoptotic stimuli , including nuclear factor ( NF ) -kappaB with its target genes IEX-3 , P04179 , P05231 , and P10145 . We demonstrate that suppression of NF-kappaB activation markedly enhances SERM-induced apoptosis , suggesting a role for NF-kappaB in protecting glioma cells from SERM-induced cytotoxicity . These findings demonstrate for the first time that a SERM other than tamoxifen can induce glioma cell apoptosis in vitro and in vivo and that the clinical efficacy of SERMs for the treatment of malignant gliomas could potentially be enhanced by simultaneous inhibition of the NF-kappaB pathway . Loss of complex I due to mitochondrial DNA mutations in renal oncocytoma . PURPOSE : Many solid tumors exhibit abnormal aerobic metabolism characterized by increased glycolytic capacity and decreased cellular respiration . Recently , mutations in the nuclear encoded mitochondrial enzymes fumarate hydratase and succinate dehydrogenase have been identified in certain tumor types , thus demonstrating a direct link between mitochondrial energy metabolism and tumorigenesis . Although mutations in the mitochondrial genome ( mitochondrial DNA , mtDNA ) also can affect aerobic metabolism and mtDNA alterations are frequently observed in tumor cells , evidence linking respiratory chain deficiency in a specific tumor type to a specific mtDNA mutation has been lacking . EXPERIMENTAL DESIGN : To identify mitochondrial alterations in oncocytomas , we investigated the activities of respiratory chain enzymes and sequenced mtDNA in 15 renal oncocytoma tissues . RESULTS : Here , we show that loss of respiratory chain complex I ( DB00157 /ubiquinone oxidoreductase ) is associated with renal oncocytoma . Enzymatic activity of complex I was undetectable or greatly reduced in the tumor samples ( n = 15 ) . Blue Native gel electrophoresis of the multisubunit enzyme complex revealed a lack of assembled complex I . Mutation analysis of the mtDNA showed frame-shift mutations in the genes of either subunit P03886 , P03905 , or P03915 of complex I in 9 of the 15 tumors . CONCLUSION : Our data indicate that isolated loss of complex I is a specific feature of renal oncocytoma and that this deficiency is frequently caused by somatic mtDNA mutations . Lowering of body core temperature by exposure to a cold environment and by a P08908 agonist : effects on physiological and psychological variables and blood serotonin levels . The present study was designed to compare the effects of a pharmacologically induced decrease in body core temperature to the effects observed with lowering of body temperature by exposure to a cold environment . Our special interest was the involvement of 5-HT in thermoregulatory responses . Sixty healthy male volunteers were randomly assigned to one of the following conditions : exposure to normal ambient temperature ( 28 degrees C ) and placebo , exposure to cold ambient temperature ( 5 degrees C ) and placebo , or normal ambient temperature and 10 mg of the partial P08908 agonist ipsapirone . As indicators of physiological responses to lowering of body temperature , tympanic temperature , skin temperature , P15941 , metabolic rate , and heart rate were monitored and saliva cortisol levels and peripheral 5-HT concentrations were determined . In addition , ratings on ambient temperature , thermal discomfort , and feelings of irritability were obtained . While lowering of body core temperature was associated with marked counterregulations ( decrease of skin temperature , increase in P15941 and metabolic rate ) and feelings of discomfort , this was not observed with ipsapirone . An increase in cortisol levels was primarily observed in the ipsapirone group and was not reflected by respective changes in whole blood or platelet 5-HT indicating that brain and platelet 5-HT are not related .

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Genetics of idiopathic disseminated bronchiectasis . Bronchiectasis is an abnormal dilation of bronchi , consequent to the destruction of their walls . It is included in the category of obstructive pulmonary diseases , along with chronic obstructive pulmonary disease ( P48444 ) , asthma , and cystic fibrosis . In approximately 50 % of cases , bronchiectasis is associated with underlying conditions ; in the remainder , known causes are not ascertainable ( idiopathic bronchiectasis ) . A search for genetic determinants of this phenotype , with the cystic fibrosis gene as a candidate , has been performed by three independent groups . The results of this search agreed on the association of bronchiectasis with cystic fibrosis gene mutations and polymorphisms . The cystic fibrosis gene is also associated with bronchiectasis due to rheumatoid arthritis and allergic bronchopulmonary aspergillosis . A few other genes have been investigated in idiopathic bronchiectasis , with negative results . Idiopathic bronchiectasis is , therefore , to be considered as an obstructive multifactorial disorder belonging to the category of cystic fibrosis monosymptomatic diseases ( or P13569 -opathies ) , whose pathogenesis is influenced by environmental factors and other undetermined genes . Oxidative stress induces extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase in cystic fibrosis lung epithelial cells : Potential mechanism for excessive P10145 expression . Cystic fibrosis ( CF ) is a lethal disease caused by defective function of the cftr gene product , the CF transmembrane conductance regulator ( P13569 ) that leads to oxidative damage and excessive inflammatory response in lungs of CF patients . We here report the effects of oxidative stress ( hyperoxia , 95 % O(2) ) on the expression of pro-inflammatory interleukin ( IL ) -8 and P25024 /2 receptors in two human CF lung epithelial cell lines ( IB3-1 , with the heterozygous F508del/W1282X mutation and CFBE41o- with the homozygous F508del/F508del mutation ) and two control non-CF lung epithelial cell lines ( S9 cell line derived from IB3-1 after correction with wtCFTR and the normal bronchial cell line 16HBE14o- ) . Under oxidative stress , the expression of P10145 and P25024 /2 receptors was increased in CF , corrected and normal lung cell lines . The effects of oxidative stress were also investigated by measuring the transcription nuclear factor kappaB ( NF-kappaB ) and activator protein-1 ( AP-1 ) activities . Under oxidative stress , no increase of NF-kappaB activation was observed in CF lung cells in contrast to that observed in normal and corrected CF lung cells . The signalling of mitogen-activated protein ( Q96HU1 ) kinases was further studied . We demonstrated that extracellular signal-regulated kinase ( P27361 /2 ) and AP-1 activity was markedly enhanced in CF but not non-CF lung cells under oxidative stress . Consistently , inhibition of P27361 /2 in oxidative stress-exposed CF lung cells strongly decreased both the P10145 production and P25024 /2 expression . Therefore , targeting of P27361 /2 Q96HU1 kinase may be critical to reduce oxidative stress-mediated inflammation in lungs of CF patients . 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 , is a selective , non-competitive inhibitor of acetylcholinesterase ( P22303 ) . In vivo , DB04892 produces rapid , potent , and long-lasting P22303 inhibition . As a possible result of its preferential brain selectivity , DB04892 is significantly less toxic than (-)-physostigmine . In studies using the Stone maze paradigm , DB04892 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 appears to have a second mode of action . Reduced secretion of beta-amyloid ( Abeta ) has been observed in cell lines exposed to DB04892 , occurring through translational regulation of beta-amyloid precursor protein ( beta- P05067 ) 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 treatment tended to reduce beta- P05067 and Abeta levels in plasma samples . Clinical studies also reveal that DB04892 ( 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 in patients with AD are currently ongoing . Ca2+ response of rat mesangial cells to DB00171 analogues . The aim of this investigation was to characterise the effects of DB00171 analogues and UTP on the single cell intracellular Ca2+ concentration ( [Ca2+]i ) in cultured rat mesangial cells . Typically , there were two phases in the Ca2+ response to the agonists , an initial fast transient peak and a subsequent slower decline , or plateau , phase . For the peak amplitude in [Ca2+]i the agonists had about equal effect . But when taking in consideration the percentage of responding cells and the integrated Ca2+ response over 1 min , the order of efficacy of nucleotide agonists ( 100 microM ) was UTP = DB00171 > ATPgammaS > ADP = 2MeS- DB00171 ( 2-methylthio- DB00171 ) . DB00640 , AMP and beta,gamma-Me- DB00171 ( 100 microM ) had no effect . DB04786 ( 100 microM ) and reactive blue ( 50 microM ) decreased the number of responding cells . Removing Ca2+ from the bath diminished neither the peak in [Ca2+]i nor the percentage of responding cells , but the average [Ca2+]i increase in 1 min was significantly reduced . The results indicate that P41231 receptors are present in rat mesangial cells but it can not be excluded that there are receptors distinct from P41231 which also mediate a rise in [Ca2+]i . DB08901 may overcome resistance of P36888 -ITD harbouring additional point mutations , notably the previously refractory F691I mutation . Fms-like tyrosine kinase ( P36888 ) mutations are the most frequent mutations in patients with acute myeloid leukaemia ( AML ) that confer a poor prognosis . Constitutively active P36888 -ITD ( internal tandem duplications ) mutations define a promising target for therapeutic approaches using small molecule inhibitors . However , several point mutations of the P36888 tyrosine kinase domain ( P36888 -TKD ) have been identified to mediate resistance towards P36888 tyrosine kinase inhibitors ( P36888 -TKI ) , including secondary mutations of P36888 . We investigated the cellular effects of the recently characterised P36888 -TKI ponatinib ( DB08901 ) on murine myeloid cells transfected with P36888 -ITD with or without additional point mutations of the P36888 -TKD including the ( so far ) multi-resistant F691I mutation . DB08901 effectively induced apoptosis not only in the parental P36888 -ITD cell line but also in all stably transfected subclones harbouring additional P36888 -TKD point mutations ( N676D , F691I or G697R ) . These observations correlated with a strong inhibition of P36888 -ITD and its downstream targets P42229 , AKT and P27361 /2 upon ponatinib incubation , as determined by Western blotting . We conclude that ponatinib represents a promising P36888 -TKI that should be further investigated in clinical trials . The targeted therapy of P36888 -ITD-positive AML with ponatinib might be associated with a lower frequency of secondary resistance caused by acquired P36888 -TKD mutations . Physiological regulation of DB00171 release at the apical surface of human airway epithelia . Extracellular DB00171 and its metabolite adenosine regulate mucociliary clearance in airway epithelia . Little has been known , however , regarding the actual DB00171 and adenosine concentrations in the thin ( approximately 7 microm ) liquid layer lining native airway surfaces and the link between DB00171 release/metabolism and autocrine/paracrine regulation of epithelial function . In this study , chimeric Staphylococcus aureus protein A-luciferase ( SPA-luc ) was bound to endogenous antigens on primary human bronchial epithelial ( P02100 ) cell surface and DB00171 concentrations assessed in real-time in the thin airway surface liquid ( ASL ) . DB00171 concentrations on resting cells were 1-10 nm . Inhibition of ecto-nucleotidases resulted in DB00171 accumulation at a rate of approximately 250 fmol/min/cm2 , reflecting the basal DB00171 release rate . Following hypotonic challenge to promote cell swelling , cell-surface DB00171 concentration measured by SPA-luc transiently reached approximately 1 microm independent of ASL volume , reflecting a transient 3-log increase in DB00171 release rates . In contrast , peak DB00171 concentrations measured in bulk ASL by soluble luciferase inversely correlated with volume . DB00171 release rates were intracellular calcium-independent , suggesting that non-exocytotic DB00171 release from ciliated cells , which dominate our cultures , mediated hypotonicity-induced nucleotide release . However , the cystic fibrosis transmembrane conductance regulator ( P13569 ) did not participate in this function . Following the acute swelling phase , P02100 cells exhibited regulatory volume decrease which was impaired by apyrase and facilitated by DB00171 or UTP . Our data provide the first evidence that DB00171 concentrations at the airway epithelial surface reach the range for P41231 receptor activation by physiological stimuli and identify a role for mucosal DB00171 release in airway epithelial cell volume regulation . Interplay between inhibitory ferric and stimulatory curcumin regulates phosphorylation-dependent human cystic fibrosis transmembrane conductance regulator and ΔF508 activity . Curcumin potentiates cystic fibrosis transmembrane conductance regulator ( P13569 ) activation in an DB00171 -independent but phosphorylation-dependent manner . The underlying molecular mechanisms are unclear . Here , P29320 -293T cells cultured in an Fe(3+)-containing medium were transiently transfected with P13569 constructs , and the role of the inhibitory Fe(3+) bridge between intracellular loop 3 and the regulatory domain of P13569 in this pathway was investigated . The results showed that ethylenediaminetetraacetic acid ( DB00974 ) stimulated phosphorylation-dependent P13569 activation and the stimulation was suppressed by the deletion of the regulatory domain or the insertion of a C832A mutation that removes the Fe(3+)-binding interface . Furthermore , curcumin potentiation of P13569 was significantly weakened not only by Fe(3+)-insensitive mutations at the interface between the regulatory domain and intracellular loop 3 but also by N-ethylmaleimide or DB00974 pretreatment that removes Fe(3+) . More importantly , potentiation of P13569 was completely suppressed by sufficient Fe(3+) . Finally , the insertion of Fe(3+)-insensitive H950R/S768R increased the curcumin-independent activity of ΔF508 but weakened its curcumin potentiation . Thus , Fe(3+) homeostasis in epithelia may play a critical role in regulating P13569 activity , and targeting Fe(3+)-chelating potentiators may direct new therapies for cystic fibrosis . P01375 polymorphisms as a potential modifier gene in the cystic fibrosis . Modifier genes , as the P01375 -α gene , can modulate the cystic fibrosis ( CF ) severity . Thus , -238G > A and -308G > A polymorphisms of P01375 -α gene were analyzed as modifiers of CF . In this context , the present study enrolled 49 CF patients ( diagnosis performed by sweat test and complete P13569 screening ) . The -238G > A polymorphism analysis was performed by Q9ULH0 -PCR , and -308G > A , by PCR-RFLP . In our data , the -238G > A polymorphism was not associated with clinical variability . The AA genotype for -308G > A polymorphism was a risk factor for early gastrointestinal symptoms ( OR=5.98 , 95 % CI=1.06-49.68 ) and protection for the first Pseudomonas aeruginosa ( OR=0.05 , 95 % CI=0.0003-0.007 ) . For the first P. aeruginosa , GA genotype was a risk factor ( OR=10.2 , 95 % CI=1.86-84.09 ) ; for the same genotype , the diagnosis was made in minor time than the AA genotype ( p=0.031 ) . Considering the -308G > A polymorphism alleles , the G allele was a risk factor for early pulmonary symptoms ( OR=3.81 , 95 % CI=1.13-12.97 ) and P. aeruginosa ( OR=66.77 , 95 % CI=15.18-482.7 ) ; however , the same allele showed better transcutaneous oxygen saturation ( OR=9.24 , 95 % CI=1.53-206.1 ) . The A allele was a protective factor for early pulmonary symptoms ( OR=12.26 , 95 % CI=0.08-0.89 ) and P. aeruginosa ( OR=12.15 , 95 % CI=0002-0007 ) , however , the same allele was a risk factor for worst transcutaneous oxygen saturation ( OR=7.01 , 95 % CI=1.14-157.4 ) . As conclusion , the -308G > A polymorphism of the P01375 -α gene was associated with the CF severity . DB09280 - DB08820 in Patients with Cystic Fibrosis Homozygous for Phe508del P13569 . P08246 inhibitors as treatment for P48444 . Chronic obstructive pulmonary disease , characterised by a slowly progressive , irreversible airways limitation , is a major worldwide cause of chronic morbidity and mortality . The imbalance between human neutrophil elastase and endogenous antiproteases may cause excess human neutrophil elastase in pulmonary tissues , which may be considered a major pathogenic factor in chronic obstructive pulmonary disease . Great effort has been devoted to finding a method to restore the balance , resulting in the discovery of potent two-typed small-molecular-weight human neutrophil elastase inhibitors . In the application of chronic obstructive pulmonary disease therapy , the human neutrophil elastase inhibitors mainly focused upon include ONO-5046 , MR-889 , L-694,458 , CE-1037 , GW-311616 and TEI-8362 as the acyl-enzyme inhibitors ; and DB03925 , AE-3763 , FK-706 , ICI-200,880 , ZD-0892 and ZD-8321 as the transition-state inhibitors . In this review , various problems that remain to be solved in the clinical use of human neutrophil elastase inhibitors are discussed . Managing the underlying cause of cystic fibrosis : a future role for potentiators and correctors . Cystic fibrosis ( CF ) , a severe genetic disease , is caused by mutations that alter the structure and function of P13569 , a plasma membrane channel permeable to chloride and bicarbonate . Defective anion transport in CF irreversibly damages the lungs , pancreas , liver , and other organs . CF mutations cause loss of P13569 function in multiple ways . In particular , class 3 mutations such as p.Gly551Asp strongly decrease the time spent by P13569 in the open state ( gating defect ) . Instead , class 2 mutations impair the maturation of P13569 protein and its transport from the endoplasmic reticulum to the plasma membrane ( trafficking defect ) . The deletion of phenylalanine 508 ( p.Phe508del ) , the most frequent mutation among CF patients ( 70-90 % ) , destabilizes the P13569 protein , thus causing both a trafficking and a gating defect . These two defects can be overcome with drug-like molecules generically called correctors and potentiators , respectively . The potentiator Kalydeco™ ( also known as DB08820 or VX-770 ) , developed by Vertex Pharmaceuticals , has been recently approved by the US FDA and the European Medicines Agency ( P15941 ) for the treatment of CF patients carrying at least one P13569 allele with the p.Gly551Asp mutation ( 2-5 % of all patients ) . In contrast , the corrector VX-809 , which significantly improves p.Phe508del- P13569 trafficking in vitro , is still under study in clinical trials . Because of multiple defects caused by the p.Phe508del mutation , it is probable that rescue of the mutant protein will require combined treatment with correctors having different mechanisms of action . This review evaluates the status of experimental and clinical research in pharmacotherapy for the CF basic defect . Haemoglobin Hope in a northern Thai family : first identification of homozygous haemoglobin Hope associated with haemoglobin H disease . Haemoglobin ( Hb ) Hope [ beta136( O43583 ) DB00145 --> DB00128 ( P19440 --> Q6IB77 ) ] is one of the unstable haemoglobin variants of the beta-globin chain , which is demonstrated in people of various ethnic backgrounds . Here we report a Thai female patient with clinical thalassaemia intermedia since childhood . This patient had experienced neither blood transfusion nor hospitalisation . Hb Bart's-H and a large amount of Hb Hope were identified by high-performance liquid chromatography ( HPLC ) assay and the diagnosis of homozygous Hb Hope was definitely achieved by direct sequencing of exon 3 of beta-globin gene . Furthermore , we could identify that her brother carried the mutation of homozygous Hb Hope without abnormal alpha globin chain involvement , and another family member had heterozygous Hb Hope in association with -alpha(3.7) mutation , and both of them were clinically silent . Genetics and treatment options for recurrent acute and chronic pancreatitis . Worldwide research efforts demonstrate a major role of gene-environment interactions for the risk , development , and progression of most pancreatic diseases , including recurrent acute and chronic pancreatitis . New findings of pancreas disease-associated risk variants have been reported in the P15085 , P19440 , P57739 , P03956 , P42898 , and other genes . These risk genes and their regulatory regions must be added to the known pathogenic variants in the P07477 , P00995 , P13569 , Q99895 , P41180 , Q8IWV7 , Q9Y3A5 , P19835 , and P07858 genes . This new knowledge promises to improve disease management and prevention through personalized medicine . At the same time , however , knowledge of an increasing number of pathogenic variants , and their complicated effects when present in combination , results in increasing difficulty in interpretation and development of recommendations . Direct-to-consumer marketing of genetic testing results also adds complexity to disease management paradigms , especially without interpretation and , in many cases , proven accuracy . While improvements in the ability to rapidly and accurately interpret complex genetic tests are clearly needed , some results , such as pathogenic P13569 variants , including a new class of bicarbonate-defective mutations , and P07477 variants have immediate implications that direct management . In addition , discovery of pancreatitis-associated genetic variants in patients with glucose intolerance may suggest underlying type 3c diabetes , which also has implications for treatment and disease management . Interpretation of point-of-care INR results in patients treated with dabigatran . BACKGROUND : Point-of-care devices for measurement of the international normalized ratio ( INR ) are commonly used to monitor therapy and maintain therapeutic levels of anticoagulation in patients treated with vitamin K antagonists . DB06695 , a new oral , reversible direct thrombin inhibitor approved for stroke prevention in patients with atrial fibrillation does not require routine coagulation monitoring . However , case reports have identified falsely elevated point-of-care INR levels in patients treated with dabigatran using one of these devices ( Hemochron ) . This in vitro study was designed to verify this issue . METHODS : We compared INR levels in whole blood and plasma using a Hemochron Jr . Signature+ point-of-care device ( International Technidyne Corporation , Edison , NJ ) with routine laboratory monitoring , using blood from healthy volunteers that was spiked with increasing concentrations of dabigatran . RESULTS : P00734 time and INR levels were increased about 2- to 4-fold with the point-of-care device compared with laboratory measures across the plasma dabigatran concentration range 50-1400 ng/mL . At plasma concentrations of dabigatran likely to be observed in patients , at a dose of 150 mg twice daily ( 60-275 ng/mL ) , whole blood point-of-care INR values increased from 1.7 to 4.0 , versus 1.1 to 1.5 measured with the laboratory coagulometer . Similar differences in prothrombin time were observed in plasma samples . CONCLUSIONS : INR levels in patients taking dabigatran are substantially higher using a Hemochron Jr. point-of-care device compared with laboratory values . We discourage the use of these devices specifically , as well as the use of the INR in general , for measuring the anticoagulant effect of dabigatran . The association between thrombophilic gene mutations and recurrent pregnancy loss . PURPOSE : To determine whether the Factor V ( 1691G/A ) , Factor V HR2 ( 4070A/G ) , P00734 ( 20210G/A ) , P05121 ( -675 I/D , 5G/4G ) , P12821 ( intron 16 I/D ) , Factor VII ( Gln353Arg ) , Factor XIII ( Val34Leu ) , β-fibrinogen ( -455G/A ) , Glycoprotein Ia ( 807C/T ) , tPA ( intron 8 D/I ) gene mutations could be risk factors for recurrent pregnancy loss ( RPL ) . METHODS : Genotyping of thrombophilic gene mutations were carried out by amplification Refractory Mutation System-PCR ( Q9ULH0 -PCR ) method after DNA extraction . RESULTS : We found that the mutant allele frequencies of Factor V ( 1691G/A ) , Factor V HR2 ( 4070A/G ) , P00734 ( 20210G/A ) , P05121 ( -675 I/D , 5G/4G ) , Factor XIII ( Val34Leu ) and β-fibrinogen ( -455G/A ) were more seen in the case group compared with the healthy control ; However , the difference between the two group is not statistically significant ( p > 0.05 ) . Whilst the mutant allele frequencies of other studied genes were lower in the case in comparison to the fertile control women ( p > 0.05 ) . CONCLUSION : Taken together , our data has shown that the prevalence of thrombophilic gene mutations was similar in women with RPL and healthy controls . Therefore , it appears that further studies on large-scale population and other genetic variants will be needed to conclusively find candidate genes for RPL unknown etiology in the future . DNA methylation and Yin Yang-1 repress adenosine A2A receptor levels in human brain . DB00640 A(2A) receptors ( A(2A) Rs ) are G-protein coupled receptors that stimulate adenylyl cyclase activity . The most A(2A) Rs-enriched brain region is the striatum , in which A(2A) Rs are largely restricted to GABAergic neurons of the indirect pathway . We recently described how DNA methylation controls basal A(2A) R expression levels in human cell lines . The present report provides clues about the molecular mechanisms that promote human brain region-specific A(2A) R gene ( P29274 ) basal expression . The transcription factors ZBP-89 and Yin Yang-1 ( P25490 ) have been characterized as regulators of P29274 in SH-SY5Y cells by means of specific expression vectors/siRNAs transient transfection and chromatin immunoprecipitation assay . ZBP-89 plays a role as an activator and P25490 as a repressor . No differences were found in ZBP-89 levels with western blot between the putamen and cerebellum of human postmortem brains . However , increased P25490 levels and DNA methylation percentage in the 5' untranslated region of P29274 , using SEQUENOM MassArray , were found in the cerebellum with respect to the putamen of human brains , showing an inverse relationship with A(2A) R levels in the two cerebral regions . Mitogen activated protein kinase 14-1 regulates serum glucocorticoid kinase 1 during seawater acclimation in Atlantic killifish , Fundulus heteroclitus . The Atlantic killifish ( Fundulus heteroclitus ) is an environmental sentinel organism used extensively for studies of environmental toxicants and osmoregulation . Previous research in our laboratory has shown that acute acclimation to seawater is mediated by an increase in O00141 . O00141 promotes the trafficking of P13569 chloride channels from intracellular vesicles to the plasma membrane of the gill within the first hour in seawater resulting in increased chloride secretion . Although we have shown that the increase in gill O00141 does not require activation of the glucocorticoid receptor , the mechanisms that mediate the rise O00141 during acute acclimation is unknown . To test the hypothesis that mitogen activated protein kinase ( Q16539 ) is responsible for the rise in O00141 we identified the coding sequence of killifish Q16539 -1 and designed a translational blocking vivo-morpholino targeting Q16539 -1 . Injection of the Q16539 -1 vivo-morpholino resulted in a 30 % reduction of Q16539 -1 and a 45 % reduction in phosphorylated- Q16539 -1 protein in the gill of killifish transitioned from freshwater to seawater . Knock down of phosphorlyated- Q16539 -1 completely blocked the rise in O00141 mRNA and protein in the killifish gill , providing the first direct and in vivo evidence that Q16539 -1 is necessary for acute seawater acclimation . A cell-based luciferase-dependent assay for the quantitative determination of free extracellular adenosine with paracrine signaling activity . Extracellular adenosine exerts powerful paracrine effects on immune cells . Thus , adenosine signaling has to be strictly regulated . This is achieved by its rapid internalization or enzymatic degradation . Consequently , free adenosine is extremely difficult to measure in cell culture systems and may escape from detection by time-consuming endpoint measurements like high-performance liquid chromatography ( HPLC ) . Therefore , we have now developed a highly sensitive assay which enables the quantification of biologically relevant extracellular adenosine via the activation of an ectopically expressed DB00640 2a-receptor ( P29274 ) in P29320 -293 reporter cells . Binding of the short-lived nucleoside to this receptor induces a DB02527 -dependent signal which can be detected via a DB02527 -responsive luciferase construct . Tests with exogenously added adenosine confirmed that the resulting luminescence signals correlate with the respective adenosine levels and thus allow quantitative measurements in a range from 20 nM to 80 μM free extracellular adenosine . Inhibition of adenosine uptake by dipyridamole further increased the sensitivity of the assay . We further validated our approach by quantifying the adenosine levels that are generated by regulatory T cells via ectonucleotidase-mediated cleavage of DB00171 . As expected , values returned to baseline when P29274 was inhibited . This confirmed that this new cell-based reporter assay constitutes a biologically relevant , technically easy , versatile , scalable and cost-effective approach that allows the non-radioactive quantification of adenosine as a signaling intermediate . Effects of P08908 and Q13639 receptor agonists on slow synaptic potentials in enteric neurons . Intracellular electrophysiological methods were used to examine the effects of 5-hydroxytryptamine ( 5-HT ) , 5-carboxamidotryptamine ( 5-CT ) , 5-methoxytryptamine ( 5-MeOT ) , 4-amino-5-chloro-2-methoxy-N-(4-[1-azabicyclo[3,3,1]nonyl]) benzamide hydrochloride ( renzapride ) , cis-4-amino-5-chloro-N [ 1- [ 3- (4-fluorophenoxy)propyl ] -3-methoxy-4-piperidinyl [ -2-methoxybenzamide monohydrate ( cisapride ) and endo-N-(8-methyl-8-azabicyclo[3.2.1]oct-3-yl)-2,3-dihydro-3- (1-methyl)ethyl-2-oxo-1 H-benzimidazole-1-carboxamidehydrochloride ( BIMU 8 ) on noncholineric slow excitatory postsynaptic potentials ( slow EPSPs ) in myenteric afterhyperpolarization ( AH ) neurons of guinea pig ileum . 5-HT ( 0.01-1 microM ) and 5-CT ( 0.001-0.1 microM ) produced a concentration-dependent inhibition of slow EPSPs . The P08908 receptor antagonist 1-(2-methoxyphenyl)-4- [ 4- ( 2-phthalimidobutyl ] piperazine ( NAN-190 ) produced rightward shifts in 5-HT and 5-CT concentration-response curves ; facilitation of slow EPSPs was never observed . 5-MeOT caused a depolarization and inhibited spike afterhyperpolarizations in a concentration-dependent manner but this effect was not blocked by the 5- Q9H205 / Q13639 receptor antagonist , tropisetron ( 1 microM ) . DB04917 ( 0.01-0.3 microM ) , cisapride ( 0.01-1.0 microM ) and BIMU 8 ( 0.01-1.0 microM ) did not change the membrane potential of any neuron tested . DB04917 and BIMU 8 did not change the amplitude of slow EPSPs . In 13 of 19 neurons cisapride did not change the amplitude of slow EPSPs ; in 6 neurons cisapride ( 1 microM ) reversibly inhibited the slow EPSP . Responses to DB05875 which mimicked the slow EPSP were not affected by cisapride. ( ABSTRACT TRUNCATED AT 250 WORDS ) Cellular and molecular effects of malnutrition and their relevance to periodontal diseases . In response to periodontal pathogens , the leukocytes ( PMN ) elaborate destructive oxidants , proteinases , and other factors . The balance between these factors , the antioxidants and endogenously synthesized antiproteinases determine the extent of periodontal damage . Malnutrition ( P15941 ) is characterized by marked tissue depletion of the key antioxidant nutrients , including DB00143 ( gamma-glutamyl-cysteinyl-glycine ) , and impaired acute-phase protein response ( Q07954 ) to infections . The latter results in diminished production of the acute-phase proteins ( P05067 ) . The Q07954 plays a key role in promoting healing , and its deficit in P15941 is due to impairment in the production and cellular action of the cytokines . Other features of malnutrition include inverted helper-suppressor T-cell ratio , histaminemia , hormonal imbalance with increased blood and saliva levels of free cortisol , and defective mucosal integrity . Malnutrition , particularly of the P15941 type which usually involves concomitant deficiencies of several essential macro- and micronutrients , therefore has the potential to adversely influence the prognosis of periodontal infections . PP2Cdelta ( Ppm1d , O15297 ) , an endogenous inhibitor of p38 MAPK , is regulated along with Trp53 and Cdkn2a following p38 MAPK inhibition during mouse preimplantation development . Preimplantation embryos utilize mitogen-activated protein kinase signaling ( MAPK ) pathways to relay signals from the external environment to prepare appropriate responses and adaptations to a changing milieu . It is therefore important to investigate how MAPK pathways are regulated during preimplantation development . This study was conducted to investigate whether PP2Cdelta ( Ppm1d , O15297 ) is expressed during mouse preimplantation development and to determine the influences of p38 MAPK inhibition on expression of Trp53 ( p53 ) , Ppm1d , ( O15297 ) , and Cdkn2a ( p16 ) during mouse preimplantation development . Our results indicate that Trp53 , Ppm1d , and Cdkn2a mRNAs and TRP53 and PP2Cdelta proteins are expressed throughout mouse preimplantation development . Treatment of 2-cell embryos with DB04338 ( potent inhibitor of p38 MAPK alpha/beta/ Q16539 /11 ) significantly increased Trp53 , Ppm1d and Cdkn2a and Mapk14 mRNA levels at 12 and 24 hr . Treatment of 8-cell embryos with DB04338 for 12 hr increased Trp53 , Ppm1d , and Cdkn2a mRNA levels , but not Mapk14 mRNA levels . Treatment of 8-cell embryos for 24 hr increased Trp53 , and Ppm1d mRNA levels , but decreased Cdkn2a and Mapk14 mRNA levels . Therefore , blockade of p38 MAPK activity is associated with embryo stage specific influences on Trp53 , Ppm1d , Cdkn2a , and Mapk14 expression during mouse preimplantation development . These results define downstream targets of p38 MAPK during preimplantation development and indicate that the p38 MAPK pathway regulates Trp53 , Ppm1d , and Cdkn2a expression . This study increases our understanding of the mechanisms controlling preimplantation development and of the interactions between preimplantation embryos and their culture environments . 5-Hydroxytryptamine contributes significantly to a reflex pathway by which the duodenal mucosa protects itself from gastric acid injury . Although duodenal mucosal bicarbonate secretion ( DMBS ) is currently accepted as an important defense mechanism against acid-induced duodenal injury , the mechanism and the regulation of DMBS are largely unknown . 5-HT may regulate DMBS , but little is known about its physiological relevance in DMBS and the underlying mechanism(s) . Thus , the aims of the present study were to demonstrate the role of 5-HT in acid-stimulated DMBS and to further elucidate the precise mechanisms involved in this process . DB01174 acid stimulation significantly increased 5-HT release from the duodenal mucosa ( P < 0.01 ) . SB204070 , a selective Q13639 receptor antagonist , dose-dependently reduced luminal acid-stimulated HCO3(-) secretion of mice in vivo . In Ussing chamber studies , 5-HT-induced I(SC) and DMBS were abolished by removal of extracellular Ca2+ , and significantly attenuated by pharmacological blockade of the Na+/Ca2+ exchanger ( O43763 ) , intermediate Ca2+-activated K+ channels ( IK(Ca) ) , or cystic fibrosis transmembrane conductance regulator ( P13569 ) . 5-HT increased cytoplasmic free calcium ( [Ca2+]cyt ) in SCBN cells , a duodenal epithelial cell line , and knockdown of P32418 proteins with a specific siRNA greatly decreased this 5-HT-mediated Ca2+ signaling . Taken together , our data suggest that 5-HT plays a physiological role in acid-stimulated DMBS via a Ca2+ signaling pathway , in which the plasma membrane O43763 transporter as well as IK(Ca) and P13569 channels may be involved .

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Mechanism of interaction of niflumic acid with heterologously expressed kidney CLC-K chloride channels . CLC-K Cl(-) channels belong to the CLC protein family . In kidney and inner ear , they are involved in transepithelial salt transport . Mutations in ClC-Kb lead to Bartter 's syndrome , and mutations in the associated subunit barttin produce Bartter 's syndrome and deafness . We have previously found that 3-phenyl-CPP blocks hClC-Ka and rClC- P04264 from the extracellular side in the pore entrance . Recently , we have shown that niflumic acid ( DB04552 ) , a nonsteroidal anti-inflammatory fenamate , produces biphasic behavior on human CLC-K channels that suggests the presence of two functionally different binding sites : an activating site and a blocking site . Here , we investigate in more detail the interaction of DB04552 on CLC-K channels . Mutants that altered block by 3-phenyl-2-(p-chlorophenoxy)propionic acid ( CPP ) had no effect on DB04552 block , indicating that the inhibition binding site of DB04552 is different from that of 3-phenyl-CPP and flufenamic acid . Moreover , DB04552 does not compete with extracellular Cl(-) ions , suggesting that the binding sites of DB04552 are not located deep in the pore . Differently from ClC-Ka , on the rat homologue P51800 , DB04552 has only an inhibitory effect . We developed a quantitative model to describe the complex action of DB04552 on ClC-Ka . The model predicts that ClC-Ka possesses two DB04552 binding sites : when only one site is occupied , DB04552 increases ClC-Ka currents , whereas the occupation of both binding sites leads to channel block . DB01257 , a terminal complement inhibitor , improves anaemia in patients with paroxysmal nocturnal haemoglobinuria . In paroxysmal nocturnal haemoglobinuria ( PNH ) , chronic destruction of PNH red blood cells ( RBCs ) by complement leads to anaemia and other serious morbidities . DB01257 inhibits terminal complement-mediated PNH RBC destruction by targeting P01031 . In the phase III , double-blind , placebo-controlled , TRIUMPH study , eculizumab reduced haemolysis , stabilized haemoglobin levels , reduced transfusion requirements and improved fatigue in patients with PNH . Herein , we explored the effects of eculizumab on measures of anaemia in patients from the TRIUMPH study and the open-label SHEPHERD study , a more heterogeneous population . DB01257 reduced haemolysis regardless of pretreatment transfusion requirements and regardless of whether or not patients became transfusion-dependent during treatment ( P < 0.001 ) . Reduction in haemolysis was associated with increased PNH RBC counts ( P < 0.001 ) while reticulocyte counts remained elevated . DB01257 -treated patients demonstrated significantly higher levels of haemoglobin as compared with placebo in TRIUMPH and relative to baseline levels in SHEPHERD ( P < 0.001 for each study ) . DB01257 lowered transfusion requirement across multiple pretreatment transfusion strata and eliminated transfusion support in a majority of both TRIUMPH and SHEPHERD patients ( P < 0.001 ) . Patients who required some transfusion support during treatment with eculizumab showed a reduction in haemolysis and transfusion requirements and an improvement in fatigue . DB01257 reduces haemolysis and improves anaemia and fatigue , regardless of transfusion requirements . Anti-stress effect of astragaloside IV in immobilized mice . ETHNOPHARMACOLOGICAL RELEVANCE : Astragaloside IV , a major component extracted from the roots of Astragalus membranaceus ( AM ) , possesses anti-inflammatory , anti-oxidative , anti-fibrotic , anti-infarction and immunoregulatory effects . To clarify anti-stress effect of AM , anxiolytic and anti-inflammatory effects of 80 % ethanol extract of AM and astragaloside IV were investigated in immobilization stress model . MATERIALS AND METHODS : The mice were orally administered with AM ( 50 , 200 , and 500 mg/kg ) , astragaloside IV ( 5 , 10 , and 20 mg/kg ) and buspirone , a positive drug , 1h before immobilization treated for 2h . For anxiolytic activity assay , EPM test was performed in mice . For anti-inflammatory activity assay , serum levels of corticosterone , P05231 and P01375 -α were measured using ELISA kits . RESULTS : AM extract and astragaloside IV increased dose-dependently time spent on open arms and open arm entries in the EPM test . Anxiolytic effects of AM extract ( 500 mg/kg ) and astragaloside IV ( 20 mg/kg ) were comparable to those of buspirone ( 1 mg/kg ) . Their anxiolytic effects were blocked by WAY-100635 ( 0.5 mg/kg , i.p. ) , a P08908 receptor antagonist ( p < 0.01 ) , but not by flumazenil ( 3 mg/kg , i.p. ) and bicuculline ( 0.5 mg/kg , i.p. ) , GABAA receptor antagonists . AM extract and astragaloside IV also reduced serum levels of corticosterone , P05231 and P01375 -α dose-dependently . CONCLUSIONS : AM , particularly astragaloside IV , may ameliorate immobilized stress-induced anxiety and inflammation . Selective targeting of the repressive transcription factors P25490 and cMyc to disrupt quiescent human immunodeficiency viruses . Quiescent HIV-1 infection of resting P01730 (+) T cells is an obstacle to eradication of HIV-1 infection . These reservoirs are maintained , in part , by repressive complexes that bind to the HIV-1 long terminal repeat ( LTR ) and recruit histone deacetylases (HDACs). cMyc and P25490 are two transcription factors that are recruited as part of well-described , distinct complexes to the HIV-1 LTR and in turn recruit HDACs . In prior studies , depletion of single factors that recruit Q13547 in various cell lines was sufficient to upregulate LTR activity . We used short hairpin RNAs ( shRNAs ) to test the effect of targeted disruption of a single transcription factor on quiescent proviruses in T cell lines . In this study , we found that depletion of P25490 significantly increases mRNA and protein expression from the HIV-1 promoter in some contexts , but does not affect Q13547 , Q92769 , O15379 , or acetylated histone 3 occupancy of the HIV-1 LTR . Conversely , depletion of cMyc or cMyc and P25490 does not significantly alter the level of transcription from the LTR or affect recruitment of HDACs to the HIV-1 LTR . Furthermore , global inhibition of HDACs with the HDAC inhibitor suberoylanilide hydroxamic acid ( DB02546 ) enhanced the increase in LTR transcription in cells that were depleted of P25490 .These findings show that despite prior isolated findings , redundancy in repressors of HIV-1 LTR expression will require selective targeting of multiple restrictive mechanisms to comprehensively induce the escape of quiescent proviruses from latency . Synthesis and biological evaluation of novel oxindole-based RTK inhibitors as anti-cancer agents . Given that receptor tyrosine kinases ( RTKs ) have emerged as key regulators of all aspects of cancer development , including proliferation , invasion , angiogenesis and metastasis , the RTK family represents an important therapeutic target for anti-cancer drug development . Oxindole structure has been used in RTK inhibitors such as DB02058 and intedanib . In this study , two series of new heterocyclic compounds containing oxindole scaffold have been designed and synthesized , and their inhibitory activity against the proliferation of nine cancer cell lines has been evaluated . Among them , compounds 9a and 9b displayed the strongest anti-proliferative activity with the IC50s below 10μM . Flow cytometric analysis showed that the compounds 9a and 9b dose-dependently arrested the cell cycle at G0/ P55008 phase . Although the leading compounds DB02058 and intedanib targets P11362 , the kinase activity test revealed that these compounds only showed slight inhibitory activity on P11362 kinase . Further enzymatic test aided by molecular docking simulation in the DB00171 -binding site demonstrated that 9a and 9b are potent inhibitors of c-Kit kinase . These compounds are worthy of further evaluation as anticancer agents . Differential recruitment of coregulator proteins steroid receptor coactivator-1 and silencing mediator for retinoid and thyroid receptors to the estrogen receptor-estrogen response element by beta-estradiol and 4-hydroxytamoxifen in human breast cancer . P03372 ( ER ) -alpha and Q92731 function as transcription factors , and both interact with nuclear regulatory proteins to enhance or inhibit transcription . We hypothesized that coregulators are expressed in breast cancer and may be differentially recruited by ERs in the presence of estrogen and tamoxifen . Q92731 was found to be expressed more frequently in node-negative patients ( P < 0.05 ) . Expression of steroid receptor coactivator-1 ( Q15788 ) was associated with nodal positivity ( P < 0.05 ) and resistance to endocrine treatment ( P < 0.001 ) . The spatial coexpression of P03372 , Q92731 , and the coregulatory proteins was established using immunofluorescence . In both cell lines ( MCF-7 and T47D ) and in primary breast cancer cell cultures , beta-estradiol up-regulated Q92731 and coregulator protein expression and increased P03372 / Q92731 interaction with the estrogen response element ( ERE ) . 4- Hydroxy-tamoxifen ( DB04468 ) increased P03372 and silencing mediator for retinoid and thyroid receptors ( Q9Y618 ) expression and increased ER-ERE binding . Q15788 and Q9Y618 were identified at the ER-ERE complex , and interactions between ER isoforms and coregulatory proteins were determined using immunoprecipitation . Both P03372 and Q92731 preferentially bound Q15788 in the presence of beta-estradiol . Conversely , in cells treated with DB04468 , P03372 and Q92731 bound Q9Y618 . Differential recruitment of Q15788 and Q9Y618 by P03372 and Q92731 in the presence of beta-estradiol and DB04468 may be central to the response of the tumor to endocrine treatment . Dual ligands targeting dopamine D2 and serotonin P08908 receptors as new antipsychotical or anti-Parkinsonian agents . Psychiatric disorders like schizophrenia and neurodegenerative diseases like Parkinson 's disease are associated with poly-factorial pathogenic mechanisms , with several neurotransmitter systems closely involved . In addition to the cerebral dopaminergic ( DA ) system , the serotoninergic ( 5-HT ) system also plays a crucial role in regulating psychoemotional , cognitive and motor functions in the central nervous system ( CNS ) . Among the large 5-HT receptor family , accumulating data have revealed new insights into the therapeutic benefit of the P08908 receptor in treating various CNS disorders , especially schizophrenia and Parkinson 's disease . The present review discusses the advance of dual agents with mixed actions at the dopamine D2 and serotonin P08908 receptors in the treatment of these diseases . Aripiprazole was the only marketed drug with dual D2 and P08908 profile . It is a partial D2 and P08908 receptor agonist and has been prescribed as an atypical antipsychotical drug . Two other drugs DB06016 and Pardoprunox are being investigated in clinic . Most of the other candidate compounds , including DB04888 , Sarizotan , Mazapertine succinate , PF-217830 , and Adoprazine were discontinued due to either non-optimal pharmacokinetic properties or insufficient therapeutical efficacy . Although much effort has been done to highlight the advantages of the P08908 and D2 dual approach , it has to be pointed out that many of these drugs showed poly-pharmacological profile by targeting many other receptors and/or transporters besides the D2 and P08908 receptors . In this regard , ' pure ' compounds exclusively acting on the D2 and P08908 receptors are highly needed to further validate this approach . Meanwhile , safety concerns and in vivo pharmacokinetic alerts should also be implanted to the drug design art early . Combined effects of C225 and 125-iodine seed radiation on colorectal cancer cells . BACKGROUND : To characterize the effect of combined treatment of the anti-epidermal growth factor receptor ( P00533 ) monoclonal antibody C225 and 125-iodine ( 125I ) seed radiation in human colorectal cancer . METHODS : We treated LS180 cells with 125I continuous low dose rate radiation in the presence and absence of 100 nM C225 . The clonogenic capacity , cellular proliferation , cell cycle distribution , apoptosis , and molecular pathways of the cells following the treatments were analyzed in vitro . RESULTS : The sensitizer enhancement ratio of C225 was approximately 1.4 . Treatment with C225 and radiation alone produced significant inhibition of cell growth , but combination therapy produced greater inhibition than either treatment administered alone . C225 increased the radiation-induced apoptosis and the fraction of γ- P16104 foci positive cells at 48 h after treatment . The Akt phosphorylation level was lower in the cells receiving the combination treatment than in the cells treated with radiation or C225 alone . CONCLUSIONS : These findings indicate that C225 sensitizes LS180 cells to 125I seed radiation . Growth inhibition is mediated by inducing apoptosis and not cell cycle arrest . Additionally , we confirmed that C225 impairs DNA repair by reducing the cellular level of the P78527 and P12956 proteins . Furthermore , the inhibition of Akt signaling activation may be responsible for the C225-mediated radiosensitization . The establishment and characterization of cell lines stably expressing human P13010 tagged with enhanced green fluorescent protein . The Ku protein is a complex of two subunits , P12956 and P13010 , and it plays a role in multiple nuclear processes , e.g. , nonhomologous DNA-end-joining ( NHEJ ) , chromosome maintenance , and transcription regulation . On the other hand , several studies have reported a cytoplasmic or cell surface localization of Ku in various cell types . The mechanism underlying the regulation of all the diverse functions of Ku is still unclear , though the mechanism that regulates the nuclear localization of P12956 and P13010 appears to play , at least in part , a key role in regulating the physiological function of Ku . In this study , we generated cell lines expressing the human P13010 tagged with the green fluorescent protein ( GFP ) color variants in P13010 -deficient cells , i.e. , xrs-6 derived from CHO- P04264 . Although P12956 , as well as P13010 , was undetectable in xrs-6 cells , it was seen in these transformants at a level similar to the level of CHO- P04264 . Furthermore , etoposide- and radiosensitive phenotype of xrs-6 cells were corrected by an introduction of the tagged P13010 . Moreover , the tagged P13010 suppressed apoptosis triggered by DNA damage . These results demonstrate that fusion to the GFP color variants does not interfere with the functions of the P13010 in the Ku-dependent DSB repair . Therefore , these transformants might be useful not only in the analysis of P13010 behavior , but also in an analysis of the dynamics of the NHEJ repair process . Histone deacetylase inhibitors increase microRNA-146a expression and enhance negative regulation of interleukin-1β signaling in osteoarthritis fibroblast-like synoviocytes . OBJECTIVE : MiR-146a exerts negative control on inflammatory responses by suppressing cytokine-induced expression of interleukin-1 receptor-associated kinase-1 ( P51617 ) and tumor necrosis factor receptor-associated factor 6 ( Q9Y4K3 ) by impairing NF-κB activity and inhibiting the expression of target genes . Recent study suggests that histone deacetylases ( HDACs ) are involved in the regulation of microRNA ( miRNA ) expression . Therefore , we determined whether HDAC inhibitors can increase miR-146a expression , thereby inhibiting interleukin-1β ( IL-1β ) -induced signaling in osteoarthritis fibroblast-like synoviocytes ( OA-FLS ) . METHOD : MiRNA expression was analyzed using real-time PCR . IL-1β-induced downstream signals and cytokine expression were evaluated using Western blotting and ELISA . Transcription factors regulating promoter activation were identified using chromatin immunoprecipitation assays . RESULTS : IL-1β treatment of OA-FLS induced a mild ( 1.7-fold ) increase in miR-146a expression that was unable to appropriately downregulate P51617 and Q9Y4K3 expression . HDAC inhibitors , DB02546 ( vorinostat ) , and LBH589 ( DB06603 ) significantly ( 6.1- and 5.4-fold ) elevated miR-146a expression by increasing the binding of the transcription factor NF-κB to the miR-146a promoter , and negatively regulated IL-1β-induced IKK/IκB/p65 phosphorylation signaling and P05231 secretion . The increase in miR-146a expression induced by the HDAC inhibitors was prevented by transfection of miR-146a inhibitor or Q13547 ( class I HDAC ) , P56524 ( class IIa HDAC ) , and Q9UBN7 ( class IIb HDAC ) overexpression , suggesting that they were due to inhibition of HDAC activity . CONCLUSIONS : Our study demonstrated that HDAC inhibitor treatment in OA-FLS significantly increased miR-146a expression and mediated markedly negative regulation to inhibit IL-1β-induced signaling and cytokine secretion . Our results indicate the potential rationale of anti-inflammatory effects for HDAC inhibitors . T cell functions in granulocyte/macrophage colony-stimulating factor deficient mice . Immunological functions were analyzed in mice lacking granulocyte/macrophage colony-stimulating factor ( GM- P04141 ) . The response of splenic T cells to allo-antigens , anti-mouse CD3 mAb , interleukin 2 ( P60568 ) , or concanavalin A was comparable in GM- P04141 +/+ and GM- P04141 -/- mice . To investigate the responses of CD8(+) and P01730 + T cells against exogenous antigens , mice were immunized with ovalbumin peptide or with DB05299 ( KLH ) . Cytotoxic CD8+ T cells with specificity for ovalbumin peptide could not be induced in GM- P04141 -/- mice . After immunization with KLH , there was a delay in IgG generation , particularly IgG2a , in GM- P04141 -/- mice . Purified P01730 + T cells from GM- P04141 -/- mice immunized with KLH showed impaired proliferative responses and produced low amounts of interferon-gamma ( P01579 ) and P05112 when KLH-pulsed B cells or spleen cells were used as antigen presenting cells ( P25054 ) . When enriched dendritic cells ( DC ) were used as P25054 , P01730 + T cells from GM- P04141 -/- mice proliferated as well as those from GM- P04141 +/+ mice and produced high amounts of P01579 and P05112 . To analyze the rescue effect of DC on P01730 (+) T cells , supernatants from ( i ) P01730 (+) T cells cultured with KLH-pulsed DC or ( ii ) DC cultured with recombinant GM- P04141 were transferred to cultures of P01730 (+) T cells and KLH-pulsed spleen cells from GM- P04141 -/- mice . Supernatants from both DC sources contained a factor or factors that restored proliferative responses and P01579 production of P01730 (+) T cells from GM- P04141 -/- mice . Total synthesis of the bicyclic depsipeptide HDAC inhibitors spiruchostatins A and B , 5''-epi-spiruchostatin B , FK228 ( FR901228 ) and preliminary evaluation of their biological activity . The bicyclic depsipeptide histone deacetylase ( HDAC ) inhibitors spiruchostatins A and B , 5''-epi-spiruchostatin B and FK228 were efficiently synthesized in a convergent and unified manner . The synthetic method involved the following crucial steps : i ) a Julia-Kocienski olefination of a 1,3-propanediol-derived sulfone and a L- or D-malic acid-derived aldehyde to access the most synthetically challenging unit , ( 3S or 3R,4E ) -3-hydroxy-7-mercaptohept-4-enoic acid , present in a D-alanine- or D-valine-containing segment ; ii ) a condensation of a D-valine-D-cysteine- or D-allo-isoleucine-D-cysteine-containing segment with a D-alanine- or D-valine-containing segment to directly assemble the corresponding seco-acids ; and iii ) a macrocyclization of a seco-acid using the Shiina method or the Mitsunobu method to construct the requisite 15- or 16-membered macrolactone . The present synthesis has established the P01031 '' stereochemistry of spiruchostatin B . In addition , HDAC inhibitory assay and the cell-growth inhibition analysis of the synthesized depsipeptides determined the order of their potency and revealed some novel aspects of structure-activity relationships . It was also found that unnatural 5''-epi-spiruchostatin B shows extremely high selectivity ( ca. 1600-fold ) for class I Q13547 ( IC(50)=2.4 nM ) over class II Q9UBN7 ( IC(50)=3900 nM ) with potent cell-growth-inhibitory activity at nanomolar levels of IC(50) values . P0DMS8 is a critical mediator in LPS-induced pulmonary inflammation . DB00640 receptor A(3) ( A(3) ) regulates directed movement of polymorphonuclear cells ( PMNs ) to sites of inflammation and has been implicated as a relevant mediator in models of inflammatory diseases . Here , we sought to characterize the role of A(3) in a murine model of lung inflammation . Initial studies revealed that pulmonary A(3) transcript levels were elevated following LPS exposure in vivo . In addition , inhalation of LPS increased the accumulation of PMNs in wild-type and A(3)(-/-) mice in all lung compartments . Pretreatment with the specific A(3)-agonist Cl- DB05511 significantly decreased migration of PMNs into lung interstitium and alveolar air space of wild-type mice but not of A(3)(-/-) mice . Lower PMN counts were associated with reduced levels of P01375 -α and P05231 in the alveolar space of wild-type mice that received Cl- DB05511 . In addition , Cl- DB05511 attenuated LPS-induced microvascular permeability in wild-type mice as assessed by the extravasation of Evans blue . In pulmonary microvascular endothelial cells , Cl- DB05511 reduced LPS-induced cytoskeletal remodeling and cell retraction , consistent with a specific role of A(3) for maintaining endothelial integrity . Migratory activity of human PMNs across an endothelial or epithelial monolayer was reduced when A(3) was activated on PMNs . Studies in chimeric mice , however , revealed that Cl- DB05511 required A(3) on both hematopoietic and nonhematopoietic cells to reduce transmigration in vivo . Together , our results shed new light on the role of A(3) in LPS-induced PMN trafficking in the lung and suggest pharmacological modulation of A(3)-dependent pathways as a promising approach in lung inflammation . Identification of an acetylation-dependant P12956 /FLIP complex that regulates FLIP expression and HDAC inhibitor-induced apoptosis . FLIP is a potential anti-cancer therapeutic target that inhibits apoptosis by blocking caspase 8 activation by death receptors . We report a novel interaction between FLIP and the DNA repair protein P12956 that regulates FLIP protein stability by inhibiting its polyubiquitination . Furthermore , we found that the histone deacetylase ( HDAC ) inhibitor DB02546 ( DB02546 ) enhances the acetylation of P12956 , thereby disrupting the FLIP/ P12956 complex and triggering FLIP polyubiquitination and degradation by the proteasome . Using in vitro and in vivo colorectal cancer models , we further demonstrated that DB02546 -induced apoptosis is dependant on FLIP downregulation and caspase 8 activation . In addition , an Q9UBN7 -specific inhibitor Tubacin recapitulated the effects of DB02546 , suggesting that Q9UBN7 is a key regulator of P12956 acetylation and FLIP protein stability . Thus , HDAC inhibitors with anti- Q9UBN7 activity act as efficient post-transcriptional suppressors of FLIP expression and may , therefore , effectively act as ' FLIP inhibitors ' . Structural basis for P01133 receptor inhibition by the therapeutic antibody DB05774 . Therapeutic anticancer strategies that target and inactivate the epidermal growth factor receptor ( P00533 ) are under intense study in the clinic . Here we describe the mechanism of P00533 inhibition by an antibody drug DB05774 . DB05774 is a fully human antibody that has similar antitumor potency as the chimeric cetuximab/Erbitux and might represent a safer therapeutic alternative . We report the X-ray crystal structure of the Fab fragment of DB05774 ( Fab11F8 ) in complex with the entire extracellular region and with isolated domain III of P00533 . We compare this to our previous study of the cetuximab/ P00533 interaction . Fab11F8 interacts with a remarkably similar epitope , but through a completely different set of interactions . Both the similarities and differences in binding of these two antibodies have important implications for the development of inhibitors that could exploit this same mechanism of P00533 inhibition . Modification of alternative messenger RNA splicing of fibroblast growth factor receptors in human cardiac allografts during rejection . Accelerated coronary atherosclerosis in cardiac transplants ( cardiac allograft vasculopathy , Q03135 ) is characterized by coronary intimal hyperplasia . P05230 ( P05230 ) is a potent mitogen for vascular smooth muscle cells and endothelial cells , and its expression is increased in cardiac allografts , suggesting it may play a role in the pathogenesis of Q03135 . The activity of P05230 is dependent on binding to transmembrane receptors . To investigate whether receptors for P05230 are also induced after transplantation , polymerase chain reaction , in situ hybridization , and immunohistochemistry were used to analyze expression of four receptors for P05230 ( P11362 - P22455 ) . Expression of mRNA encoding extracellular immunoglobulin-like domains of P11362 was increased 35-fold in cardiac allografts compared with normal hearts and was predominantly present in cardiac myocytes and vascular structures . Alternatively spliced mRNA that encodes transmembrane forms of P11362 , which contain the signal-transducing tyrosine kinase domains , was induced in allografts during rejection , in infiltrating cells , vascular structures , and myocytes . In vitro experiments showed that differential expression of FGF receptor isoforms was induced by P05230 , and also by P05231 and TGF-beta , which are expressed in cardiac allografts during rejection . The results show that expression of both P05230 and its receptors is altered in cardiac allografts and suggest that these events are important in the pathogenesis of Q03135 . Anticytokine treatment prevents the increase in the activity of DB00171 -ubiquitin- and Ca(2+)-dependent proteolytic systems in the muscle of tumour-bearing rats . The ascites hepatoma Yoshida AH-130 induces loss of body weight and tissue waste . Tumour necrosis factor alpha ( P01375 ) plays a pivotal role in the pathogenesis of muscle wasting in this model system , but other cytokines , such as interleukin-6 , may be involved . In order to verify whether a combined anticytokine treatment may synergistically counteract muscle protein degradation , tumour bearing rats were treated with pentoxyfilline ( PTX , an inhibitor of P01375 synthesis ) , or with suramin ( Q09428 , an antiprotozoal drug blocking the peripheral action of several cytokines including P05231 and P01375 ) , or both the drugs , and the effects on muscle proteolytic systems were assessed . Muscle protein loss in the AH-130-bearing rats was associated with increased activity of both the DB00171 -ubiquitin- and the calpain- dependent proteolytic pathways ( 246 % and 230 % of controls , respectively ) . Both PTX and Q09428 , either alone or in combination , prevented the depletion of muscle mass and significantly reduced the activity of muscle proteolytic systems . In particular , treatment with Q09428 , either alone or with PTX , induced a decrease in enzymatic activities to values similar to those of controls . The results obtained in the present paper demonstrate that : ( i ) muscle depletion in this model is indeed associated with increased proteasome- and calpain-dependent proteolysis , as previously suggested by increased mRNA expression of molecules pertaining to both pathways ; ( ii ) anticytokine treatments effectively reduce muscle protein loss by down-regulating the activity of at least two major proteolitic systems ; ( iii ) Q09428 is more effective than PTX in reducing the activity of proteolytic systems , possibly because of its multiple anticytokine action . Q9UGN5 interacts with Q15669 -1 and regulates expression of surfactant protein-B . P43699 ( Q15669 -1/Nkx-2.1 ) plays a critical role in lung morphogenesis and regulates the expression of lung-specific genes , including the surfactant proteins required for pulmonary function after birth . The activity of Q15669 -1 is influenced by its interactions with other transcription factors and coactivators , including CBP/p300 and Q15788 . In this study , we have identified poly(ADP-ribose) polymerases ( Q9UGN5 and P09874 ) as Q15669 -1 interacting proteins that influence its transcriptional activity . Endogenous Q9UGN5 was coimmunoprecipitated from transformed mouse lung epithelial cell ( MLE15 ) extracts with Q15669 -1 and was identified by mass spectrometry . P09874 and P12956 / P13010 were also coimmunoprecipitated from the cell extracts with Q15669 -1 . The E domain of Q9UGN5 interacted via the C-terminal domain of Q15669 -1 . Both P09874 and Q9UGN5 enhanced the activity of the promoter of surfactant protein-B ( Sftpb gene ) but not other surfactant proteins in vitro . Q9UGN5 was selectively expressed in epithelial cells of the conducting and peripheral lung tubules of the fetal mouse lung from embryonic day 12.5 and was detected in bronchial epithelial cells in the adult lung at cellular sites consistent with that of surfactant protein B . Q9UGN5 and P09874 interact with Q15669 -1 and regulate the expression of surfactant protein B , a protein required for lung function . Ontogeny of sulfonylurea-binding regulatory subunits of K( DB00171 ) channels in the pregnant rat myometrium . DB00171 -sensitive potassium channels ( K( DB00171 ) channels ) are composed of sulfonylurea receptors ( SURs ) and potassium inward rectifiers ( Kir(6.x) ) that assemble to form a large octameric channel . This study was designed to examine the expression and role of sulfonylurea-binding regulatory subunits 1 ( Q09428 ( Q09428 ) ) and 2 ( SUR2 ( O60706 ) ) of the K( DB00171 ) channels in the pregnant rat myometrium with particular regard to the contractility . RT-PCR and western blot analyses were performed to detect the presence of Q09428 and SUR2 . The Q09428 levels were markedly increased in the early stages of pregnancy . The highest level was detected on day 6 of pregnancy , whereas in the late stages , the levels of Q09428 were significantly decreased . The SUR2 level remained unchanged throughout pregnancy . The Q09428 non-selective diazoxide and the SUR2-selective pinacidil inhibited oxytocin-induced contractions . DB01016 , a K( DB00171 ) channel blocker , antagonized both pinacidil- and diazoxide-induced relaxations . It was established that SURs are responsible for pharmacological reactivity of K( DB00171 ) channel openers . We conclude that both SURs are involved in the K( DB00171 ) channel in the pregnant rat myometrium . It may further be concluded that ' pinacidil-like ' K( DB00171 ) channel openers may be of therapeutic relevance as tocolytic agents in the future .

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Regulation of Con A-dependent cytokine production from P01730 + and CD8+ T lymphocytes by autosecretion of histamine . OBJECTIVES : Previously we have shown that both P01730 + T cells and CD8+ T cells produce histamine when activated with Con A . The aim of this study was to examine whether cytokine production by these cells is regulated by autosecretion of histamine . MATERIALS : P01730 + and CD8+ T cells were separated from spleen cells of C57BL/6 mice and mice lacking the H1 receptor ( P35367 ) or P25021 , using anti- P01730 +- and anti-CD8+-coupled magnetic beads , respectively . RESULTS : Depletion of the P35367 resulted in decreases in the release of P60568 and P22301 from both P01730 + and CD8+ cells and increases in the release of P05112 from P01730 + T cells and P01579 from CD8+ cells . Mice lacking the P25021 showed up-regulation of P01579 secretion from CD8+ cells and of P05112 from P01730 + and CD8+ T cells . Release of P60568 and P22301 from P01730 + as well as CD8+ cells was down-regulated in these mice . Both P01730 + and CD8+ T cell fractions synthesized histamine , which was enhanced in the P35367 -deficient CD8+ T cells . Treatment of the cells with alpha-fluoromethyl-histidine , a specific inhibitor of HDC , or histaminase increased P01579 from CD8+ cells , whereas it had no appreciable effect on P05112 secretion from P01730 + cells . CONCLUSION : These results suggest that cytokine production by P01730 + and CD8+ T lymphocytes is regulated by autosecretion of histamine . Inhibition of P51587 and DB01643 Synthase Creates Multidrug Sensitive Tumor Cells via the Induction of Combined " Complementary Lethality " . A high mutation rate leading to tumor cell heterogeneity is a driver of malignancy in human cancers . Paradoxically , however , genomic instability can also render tumors vulnerable to therapeutic attack . Thus , targeting DNA repair may induce an intolerable level of DNA damage in tumor cells . P51587 mediates homologous recombination repair , and P51587 polymorphisms increase cancer risk . However , tumors with P51587 mutations respond better to chemotherapy and are associated with improved patient prognosis . P04818 ( TS ) is also involved in DNA maintenance and generates cellular thymidylate . We determined that antisense downregulation of P51587 synergistically potentiated drugs with mechanisms of action related to P51587 function ( cisplatin , melphalan ) , a phenomenon we named " complementary lethality. " TS knockdown induced complementary lethality to TS-targeting drugs ( 5-FUdR and pemetrexed ) but not DNA cross-linking agents . Combined targeting of P51587 and TS induced complementary lethality to both DNA-damaging and TS-targeting agents , thus creating multidrug sensitive tumors . In addition , we demonstrated for the first time that simultaneous downregulation of both targets induced combined complementary lethality to multiple mechanistically different drugs in the same cell population . In this study , we propose and define the concept of " complementary lethality " and show that actively targeting P51587 and TS is of potential therapeutic benefit in multidrug treatment of human tumors . This work has contributed to the development of a P51587 -targeting antisense oligdeoxynucleotide ( ASO ) " BR-1 " which we will test in vivo in combination with our TS-targeting ASO " Q8N1L9 83 " and attempt early clinical trials in the future.Molecular Therapy - Nucleic Acids ( 2013 ) 2 , e78 ; doi:10.1038/mtna.2013.7 published online 12 March 2013 . Pre-clinical evaluation of an in vitro selection protocol for the enrichment of transduced P28906 + cell-derived human dendritic cells . The efficient genetic modification of P28906 + cell-derived dendritic cells ( DC ) will provide a significant advancement towards the development of immunotherapy protocols for cancer , autoimmune disorders and infectious diseases . Recent reports have described the transduction of P28906 + cells via retrovirus- and lentivirus-based gene transfer vectors and subsequent differentiation into functional DC . Since there is significant apprehension regarding the clinical uses of HIV-based vectors , in this report , we compare a murine leukemia virus ( MLV ) - and a human immunodeficiency virus ( HIV ) -based bicistronic vector for gene transfer into human P28906 + cells and subsequent differentiation into mature DC . Each vector expressed both EGFP and the dominant selectable marker P00374 (L22Y) allowing for the enrichment of marked cells in the presence of the antifolate drug trimetrexate ( TMTX ) . Both MLV-based and HIV-based vectors efficiently transduced cytokine mobilized human peripheral blood P28906 + cells . However , in vitro expansion and differentiation in the presence of GM- P04141 , P01375 , Flt-3L , P21583 and P05112 resulted in a reduction in the percentage of DC expressing the transgene . Selection with TMTX during differentiation increased the percentage of marked DC , resulting in up to 79 % ( MLV vector ) and up to 94 % ( lentivirus-vector ) transduced cells expressing EGFP without loss of DC phenotype . Thus , MLV-based vectors and in vitro selection of transduced human DC show great promise for immunotherapy protocols . Optimized protocols for generation of cord blood-derived cytokine-induced killer/natural killer cells . The efficacy of various combinations of stem cell factor ( P21583 ) , P36888 ligand , interleukin ( IL ) -2 , P13232 and P40933 to induce and expand cord blood-derived cytokine-induced killer ( CIK ) cells was investigated . There were three treatment groups : group A : P21583 combined with P60568 , P13232 and P40933 ; group B : P21583 , P36888 ligand combined with P60568 , P13232 and P40933 , and group C : P60568 , P13232 and P40933 , the control group . Proliferation rates of CD3(+)CD56(+) CIK cells and CD3(-)CD56(+) natural killer ( NK ) cells were highest in group B ; expansion of CIK cells increased 796.1 ± 278.5-fold , and that of NK cells increased 36.6 ± 3.5-fold . All expanded cord blood-derived CIK/NK cells showed cytotoxic activity against the K562 cell line . Interestingly , the cytotoxicity of group A was highest and significantly higher than that of other groups . These protocols might provide an alternative choice for CIK/NK cell expansion . Paullones are potent inhibitors of glycogen synthase kinase-3beta and cyclin-dependent kinase 5/p25 . Paullones constitute a new family of benzazepinones with promising antitumoral properties . They were recently described as potent , DB00171 -competitive , inhibitors of the cell cycle regulating cyclin-dependent kinases ( CDKs ) . We here report that paullones also act as very potent inhibitors of glycogen synthase kinase-3beta ( GSK-3beta ) ( IC50 : 4-80 nM ) and the neuronal Q00535 /p25 ( IC50 : 20-200 nM ) . These two enzymes are responsible for most of the hyperphosphorylation of the microtubule-binding protein tau , a feature observed in the brains of patients with Alzheimer 's disease and other neurodegenerative ' taupathies ' . DB04014 , the most active paullone , was demonstrated to act by competing with DB00171 for binding to GSK-3beta . DB04014 inhibits the phosphorylation of tau in vivo at sites which are typically phosphorylated by GSK-3beta in Alzheimer 's disease . DB04014 also inhibits the Q00535 /p25-dependent phosphorylation of Q9UD71 in mouse striatum slices in vitro . This dual specificity of paullones may turn these compounds into very useful tools for the study and possibly treatment of neurodegenerative and proliferative disorders . A case of CPT deficiency , homoplasmic mtDNA mutation and ragged red fibers at muscle biopsy . A 45-year-old male patient had an episode of acute renal failure with myoglobinuria , myalgias , weakness , and markedly increased serum CK levels . Similar episodes had occurred in the past . DB00583 palmitoyl-transferase II ( P23786 ) deficiency was documented both biochemically and genetically . Interestingly , muscle biopsy also showed some ragged red fibers ( Q96E11 ) and complete mitochondrial DNA ( mtDNA ) sequence disclosed a homoplasmic T3394C point mutation . This mutation is described in Leber 's hereditary optic neuropathy ( LHON ) or in patients with diabetes mellitus . Immunogenicity of the Brucella melitensis recombinant ribosome recycling factor-homologous protein and its cDNA . A study was conducted to evaluate the immunogenicity of the Brucella melitensis ribosome recycling factor ( Q96E11 ) -homologous protein ( CP24 ) . The CP24 gene was cloned , expressed in Escherichia coli and purified . The resulting purified recombinant protein ( rCP24 ) produced delayed-type hypersensitivity ( DTH ) reactions in B. melitensis-infected mice but not in naive controls . Thus , we decided to characterise the immune responses generated with DNA vaccination ( pcDNACP24 ) or immunisation with the rCP24 in adjuvant . Animals injected with pcDNACP24 exhibited a dominance of IgG2a to IgG1 while mice injected with rCP24 developed a higher response of IgG1 than IgG2a . Both immunisation protocols were capable of eliciting CP24-specific gamma interferon ( P01579 ) producing cells . Spleen cells from pcDNACP24-immunised mice did not produce interleukin ( IL ) -4 , P22301 or up-regulation of P60568 mRNA . Cells from rCP24-immunised mice produced P22301 , up-regulated P60568 mRNA but did not produce P05112 . Neither immunisation with purified CP24 nor injection of pcDNACP24 protected mice against challenge with live smooth B. melitensis . However , the potential of CP24 for a Brucella diagnostic test based on an in vitro antigen ( Ag ) -specific P01579 production or DTH test would be worth testing . Reduced folate carrier and dihydrofolate reductase expression in acute lymphocytic leukemia may predict outcome : a Children 's Cancer Group Study . PURPOSE : DB00563 is a major component of current treatment regimens for children with acute lymphocytic leukemia ( ALL ) . Potential mechanisms of methotrexate resistance include impaired drug uptake , decreased drug retention , and dihydrofolate reductase ( P00374 ) amplification . The purpose of this study was to assess whether reduced folate carrier ( P41440 ) and P00374 expression in untreated leukemic blasts correlated with outcome . METHODS : Quantitative real-time RT-PCR was used to measure P41440 and P00374 mRNA expression in leukemic blasts from 40 newly diagnosed patients with ALL obtained in a blinded fashion from Children 's Cancer Group studies . RESULTS : Low P41440 expression at diagnosis correlated significantly with an unfavorable event free survival . Surprisingly , low , not high , P00374 expression correlated significantly with an unfavorable event-free survival . Proliferative cell nuclear antigen ( P12004 ) expression demonstrated a weak inverse relationship between sample P12004 and P00374 or P41440 expression , suggesting that P00374 and P41440 expression may be markers for factors other than drug resistance . CONCLUSIONS : These results suggest that impaired transport may be an important mechanism of intrinsic methotrexate resistance in ALL , and P00374 expression also may be an important prognostic factor in ALL . Additional studies are necessary to clarify the mechanism for the correlation of low P00374 expression with poor outcome . Nuclear factor kappaB ( NF-kappaB ) pathway as a therapeutic target in rheumatoid arthritis . Rheumatoid arthritis ( RA ) is a chronic inflammatory disease characterized by persistent joint swelling and progressive destruction of cartilage and bone . Current RA treatments are largely empirical in origin and their precise mechanism of action is uncertain . Increasing evidence shows that chronic inflammatory diseases such as RA are caused by prolonged production of proinflammatory cytokines including tumor necrosis factor ( P01375 ) and interleukin 1 ( IL-1 ) . The nuclear factor kappaB ( NF-kappaB ) plays an essential role in transcriptional activation of P01375 and IL-1 . NF-kappaB is induced by many stimuli including P01375 and IL-1 , forming a positive regulatory cycle that may amplify and maintain RA disease process . NF-kappaB and enzymes involved in its activation can be a target for anti-inflammatory treatment . DB00945 and sodium salicylate inhibit activation of NF-KB by blocking O15111 , a key enzyme in NF-kappaB activation . Glucocorticoids suppress expression of inflammatory genes by binding glucocorticoid receptor with NF-kappaB , and increasing expression of inhibitory protein of NF-kappaB , P25963 . Sulfasalazine and gold compounds also inhibit NF-kappaB activation . Continuing advances in our understanding of action mechanism of antirheumatic agents will benefit the future development of RA regimens with greater efficacy and less toxicity . DB00563 in pediatric osteosarcoma : response and toxicity in relation to genetic polymorphisms and dihydrofolate reductase and reduced folate carrier 1 expression . OBJECTIVE : To determine the influence of the genotype and the level of expression of different enzymes involved in folate metabolism on the response to and toxicity of high-dose methotrexate treatment in pediatric osteosarcomas . STUDY DESIGN : P00374 and Reduced folate carrier 1 ( RFC1 ) semiquantitative expression was analyzed in 34 primary and metastatic osteosarcoma tissues by real-time polymerase chain reaction . The following polymorphisms were also analyzed in peripheral blood from 96 children with osteosarcoma and 110 control subjects : C677T , A1298C ( P42898 ) , G80A ( RFC1 ) , A2756G ( Q99707 ) , C1420T ( SHMT ) , the 28bp-repeat polymorphism , and 1494del6 of the P04818 gene . Treatment toxicity was scored after each cycle according to criteria from the World Health Organization . RESULTS : P00374 and RFC1 expression was lower in initial osteosarcoma biopsy specimens than in metastases ( P = .024 and P = .041 , respectively ) . RFC1 expression was moderately decreased in samples with poor histologic response to preoperative treatment ( P = .053 ) . Patients with osteosarcoma with P46379 /G4 hematologic toxicity were more frequently TT than CT/CC for C677T/ P42898 ( P = .023 ) and GG for A2756G/ Q99707 ( P = .048 and P = .057 for gastrointestinal and hematologic toxicity , respectively ) . CONCLUSIONS : The role of C677T/ P42898 and A2756G/ Q99707 on chemotherapy-induced toxicity should be further investigated in pediatric osteosarcomas receiving high-dose methotrexate . Altered expression of P00374 and RFC1 is a feasible mechanism by which osteosarcoma cells become resistant to methotrexate . DB09559 in the treatment of advanced non-small cell lung cancer : translation from preclinical to clinical development . INTRODUCTION : Treatment outcomes in unselected patients with advanced NSCLC remain disappointing with platinum-based chemotherapy . The addition of monoclonal antibodies targeting P00533 to standard first-line therapy is a validated strategy and has been associated with statistically significant survival advantage in advanced NSCLC . Necitumunab is a fully human IgG1 monoclonal antibody targeting P00533 , having the potential benefit of lower hypersensitivity reaction risk as compared with cetuximab and also equivalent antibody-dependent cell-mediated cytotoxicity . AREAS COVERED : This paper reviews literature on preclinical and early clinical development of necitumumab that is available in PubMed and published abstracts from conferences , as well as ongoing trials as specified by clinicaltrials.gov . Recently , the Phase III clinical trial evaluating the addition of necitumumab to pemetrexed and cisplatin in non-squamous NSCLC was prematurely closed due to concerns about the increased risk of thromboembolic events in the experimental arm . Accrual in the Phase III trial of necitumumab in combination with gemcitabine and cisplatin in squamous NSCLC is ongoing . EXPERT OPINION : Results of the ongoing large randomized trials will be instrumental in determining the drug 's clinical significance and , with the analysis of potential molecular predictive factors , are expected to bring valuable additions to future therapeutic strategies in NSCLC . DB00563 in rheumatoid arthritis : studies with animal models . The present studies have shown that low doses of methotrexate can suppress the inflammation and joint destruction associated with animal models of arthritis . The antiinflammatory effects of methotrexate are probably related to its inhibitory effect on chemotaxis . At the low doses used , methotrexate does not induce systemic immunosuppression . In methotrexate-treated rats , an improvement in P60568 synthesis is observed and increases in P60568 levels are expected to improve cell mediated immunity . Suppressor cells appear to be very sensitive to methotrexate . Macrophage function is modulated by methotrexate . All of these effects including the effects on joint destruction are probably due to inhibition of P00374 activity of critical cells that are involved in the pathogenesis of rat arthritis induced either by adjuvant or by streptococcal cell walls . Some of these effects have been extended to human arthritis but additional studies are required to understand how low dose methotrexate exerts its beneficial effects in humans . Detection of thymidylate synthase modulators by a novel screening assay . P04818 ( TS ) , a key cancer chemotherapeutic target , catalyzes the conversion of deoxyuridylate to thymidylate . TS can serve as a repressor of its own synthesis by binding to its own mRNA through TS-specific binding elements ( TBEs ) . In this report , we describe the use of a luciferase reporter plasmid containing two TBEs that can be used as a tool for the identification and initial profiling of compounds that modulate TS activity , levels , or ability to bind mRNA . To validate this model , we evaluated several groups of drugs . Thus , cells were exposed to the pyrimidine analogs 5-fluorouracil ( DB00544 ) , 5-fluorouridine ( DB01629 ) , 5-fluoro-2'-deoxyuridine ( FUdR ) , trifluorothymidine ( DB00432 ) ; to the nonpyrimidine TS-inhibitors AG-331 , nolatrexed ( AG337 ) , and raltitrexed ( DB00293 ) ; or to drugs with other primary sites of action ( methotrexate , actinomycin D , 5-azacytidine , 8-thioguanosine ) . Except for 5-azacytidine and 8-thioguanosine , all compounds examined induced luciferase activity compared with untreated cells . Effects of luciferase activity inducing drugs through TS-affected translation were confirmed by examinations of TS protein and mRNA levels . Treatment of H630- P13671 cells with DB00544 , DB01629 , FUdR , DB00432 , AG331 , AG337 , DB00293 , and methotrexate up-regulated TS levels as determined by Western blot analysis , although TS mRNA levels remained unchanged as determined by reverse transcription-polymerase chain reaction . Our studies demonstrate a novel application of a TBE-dependent reporter plasmid that could be used for the high-throughput identification of potential chemotherapeutic agents that modulate TS RNA-binding activity , either directly or indirectly . Q9UEF7 gene deficiency causes salt-sensitive hypertension via monocyte chemotactic protein-1/CC chemokine receptor 2-mediated inflammation . Q9UEF7 ( KL ) is a newly discovered aging suppressor gene . In mice , the KL gene extends the lifespan when overexpressed and shortens the lifespan when disrupted . This study investigated if KL deficiency affects BP and salt sensitivity using KL mutant heterozygous ( +/- ) mice and wild-type ( WT ) mice ( 9 weeks of age , 16 mice per group ) . Notably , systolic BP in KL(+/-) mice began to increase at the age of 15 weeks , reached a peak level at the age of 17 weeks , and remained elevated thereafter , whereas systolic BP remained consistent in WT mice . High salt ( HS ) intake further increased BP in KL(+/-) mice but did not affect BP in WT mice . Blockade of CC chemokine receptor 2 ( P41597 ) , involved in monocyte chemotaxis , by a specific P41597 antagonist ( DB05130 ) abolished the HS-induced increase in BP in KL(+/-) mice . Furthermore , HS loading substantially increased the expression of monocyte chemotactic protein-1 and the infiltration of macrophages and T cells in kidneys in KL(+/-) mice , and treatment with DB05130 abolished these effects . Treatment of KL(+/-) mice with DB05130 also attenuated the increased renal expressions of serum glucocorticoid-regulated kinase 1 , thiazide-sensitive NaCl cotransporter , and DB00171 synthase β along with the renal structural damage and functional impairment induced by HS loading . In conclusion , KL deficiency caused salt-sensitive hypertension and renal damage by P41597 -mediated inflammation . P12004 -dependent kinase 5 ( Q00535 ) and neuronal cell death . Many neurological disorders like Parkinson 's and Alzheimer 's disease , amyotrophic lateral sclerosis ( P35858 ) or stroke have in common a definite loss of CNS neurons due to apoptotic or necrotic neuronal cell death . Previous studies suggested that proapoptotic stimuli may trigger an abortive and , therefore , eventually fatal cell cycle reentry in postmitotic neurons . Neuroprotective effects of small molecule inhibitors of cyclin-dependent kinases ( CDKs ) , which are key regulators of cell cycle progression , support the cell cycle theory of neuronal apoptosis . However , growing evidence suggests that deregulated Q00535 , which is not involved in cell cycle control , rather than cell cycle relevant members of the CDK family , promotes neuronal cell death . Here we summarize the current knowledge about the involvement of Q00535 in neuronal cell death and discuss possible up- or downstream partners of Q00535 . Moreover , we discuss potential therapeutic options that might arise from the identification of Q00535 as an important upstream element of neuronal cell death cascades . Copy number analysis of 24 oncogenes : O15151 identified as a putative marker for low recurrence risk in non muscle invasive bladder cancer . Patients with non-muscle invasive bladder cancer ( NMIBC ) generally have a high risk of relapsing locally after primary tumor resection . The search for new predictive markers of local recurrence thus represents an important goal for the management of this disease . We studied the copy number variations ( CNVs ) of 24 oncogenes ( O15151 , P04198 , Q9UM73 , P16234 , P10721 , P35968 , P00374 , P00533 , MET , SMO , P11362 , MYC , P00519 , P07949 , P24385 , P30279 , P11802 , Q00987 , Q96GD4 , P04626 , P11388 , O14965 , AR and P15056 ) using multiplex ligation probe amplification technique to verify their role as predictive markers of recurrence . DB03843 -fixed paraffin-embedded tissue samples from 43 patients who underwent transurethral resection of the bladder ( TURB ) were used ; 23 patients had relapsed and 20 were disease-free after 5 years . Amplification frequencies were analyzed for all genes and O15151 was the only gene that showed significantly higher amplification in non recurrent patients than in recurrent ones ( 0.65 vs. 0.3 ; Fisher 's test p=0.023 ) . Recurrence-free survival analysis confirmed the predictive role of O15151 ( log-rank test p=0.041 ) . Our preliminary results indicate a putative role for the O15151 gene in predicting local recurrence of bladder cancer . Confirmation of this hypothesis is needed in a larger cohort of NMIBC patients . Light activates reduction of methotrexate by NADPH in the ternary complex with Escherichia coli dihydrofolate reductase . DB00563 ( MTX ) , a strong inhibitor of dihydrofolate reductase ( P00374 ) , has been widely used for chemotherapy for many types of cancer as well as for juvenile rheumatoid arthritis . It mimics folate substrates and binds tightly to the active site of P00374 , perhaps in a conformation close to the transition state of the folate catalyzed reaction . Absorption , fluorescence and ultrasensitive Raman difference spectroscopies show that light-activated MTX reacts with NADPH in the enzyme active site , producing 5,8-dihydromethotrexate ( 5,8-dihydro-MTX ) and NADP+ . The reaction , which proceeds with a hydride transfer between C4 ( pro-R side ) of the nicotinamide ring and N5 of the pteridine ring , is similar to that between folate and NADPH except that the hydride is transferred to P13671 in this case . Hence , MTX is catalytically competent in its excited state . Most experiments were performed on the Escherichia coli enzyme , but preliminary studies show that the reaction also occurs with human P00374 . Flow cytometric analysis of mammalian glial cultures treated with methotrexate . DB00563 ( MTX ) is an antineoplastic drug that acts by competitive inhibition of the enzyme dihydrofolate reductase ( P00374 ) . MTX treatment of cultured cell lines leads to the emergence of resistant cell populations . Studies using stepwise selection procedures have demonstrated that MTX resistance conferred by overproduction of P00374 can be caused by P00374 gene amplification . We examined the effect of MTX on cells whose origin more closely approximates the in vivo condition by developing a culture system using dissociated brain tissue from 17-19 day old mouse embryos . At the first passage , cultures were divided into control and MTX groups . Cells were treated with the same or successively higher concentrations of MTX at each passage over a 3-4 month period . The first passage eliminated neurons and left a glial culture comprised of approximately 90 % astrocytes . We used the Fluorescence Activated Cell Sorter in conjunction with fluorescent dyes to measure P00374 content , DNA content , size , and viability of glial cells following MTX treatment . MTX-treated cells divided but grew more slowly and were larger than untreated cells . Stepwise selection in 30/60/90 nM or 60/120 nM MTX resulted in significant two- to threefold increases in fluorescence , and hence P00374 levels . Slot hybridizations assays demonstrated a threefold increase in P00374 gene copy number in the DNA from the 30/60/90 cultures . Thus , our findings were consistent with the results obtained from somatic cell lines , and lend support to the hypothesis that gene amplification may be a common mechanism for the acquisition of resistance in many types of cells . They also indicate that glial cells may be a specific target for cytotoxic effects of MTX on the central nervous system . DB00563 and cytarabine inhibit progression of human lymphoma in NOD/SCID mice carrying a mutant dihydrofolate reductase and cytidine deaminase fusion gene . An SFG-based retroviral bicistronic vector containing a double-mutant dihydrofolate reductase-cytidine deaminase fusion cDNA ( F/S P00374 -CD ) with IRES-eGFP confers resistance to both methotrexate ( MTX ) and cytarabine ( ara-C ) . Two weeks after transplantation with marrow transduced with either a fusion or a control gene ( eGFP-IRES-NeoR ) , human lymphoma ( P12755 -DLCL-1 ) cells were injected sc into the flanks of nonobese diabetic/severe combined immune deficiency mice . In mock-transplanted mice , maximal tolerated dose ( MTD ) of posttransplant MTX/ara-C ( 15/10 mg/kg/day , x3 ) was unable to control tumor growth . Transfer of the fusion gene allowed doses of MTX/ara-C ( 25/15 mg/kg/day , x4 ) twofold higher than the MTD to be tolerated . The tumor burden defined the efficiency of posttransplant chemotherapy ; early treatment , 48 h after tumor inoculation , provided tumor-free survival , while starting treatment after having palpable tumor growth ( 7 days ) delayed tumor growth a median time of 28 days . In addition , the early treated group had higher gene expression in peripheral blood and marrow cells than the late treated group ( P < 0.05 ) , suggesting that early treatment allowed for enrichment of transduced marrow progenitors . These results encourage clinical studies using this retroviral fusion gene construct . The P36888 inhibitor DB05465 ( formerly MLN518 ) has sequence-independent synergistic effects with cytarabine and daunorubicin . AML remains a difficult disease to treat . Despite response to induction chemotherapy , most patients ultimately relapse . Further , among elderly patients , aggressive therapy options are often limited due to other medical conditions and decreased tolerance of these patients to conventional chemotherapy . Internal tandem duplications ( ITD ) of the P36888 juxtamembrane domain occur in 20-30 % of AML patients and predict poor outcome . First clinical data with the P36888 inhibitor DB05465 demonstrated antileukemic activity in approximately half of the patients -- predominantly with P36888 ITD positive AML . But the data also show that optimal use of DB05465 will require combination therapy with cytotoxic agents . Notably , single agent DB05465 has not been associated with myelosuppression , mucositis or cardiac toxicity -- the dose limiting toxicities of AML chemotherapy . We determined the feasibility of combining DB05465 with the standard " 3 + 7 " induction regimen in AML and show that , in contrast to other structurally unrelated P36888 inhibitors recently evaluated in clinical trials , the use of DB05465 displayed application sequence independent synergistic antileukemic effects in combination with cytarabine and daunorubicin . Strong synergistic antiproliferative and proapoptotic effects were thereby predominantly seen on P36888 ITD positive blasts . Further we demonstrate , that addition of DB05465 may allow dose reduction of chemotherapy without loss of overall antileukemic activity -- resulting in a potential decrease of side effects . This approach might be an interesting novel strategy especially in the treatment of elderly/comorbid patients . Our data provide a rationale for combining DB05465 with induction chemotherapy in intensified as well as in dose reduction protocols for P36888 ITD positive AML . DB00134 -dependence phenotype in the de novo pathway in P38398 and P51587 mutation carriers with and without breast cancer . DB00134 -dependence phenotype ( P16444 ) refers to the reduced ability of cells to proliferate when methionine is restricted and/or replaced by its immediate precursor homocysteine . P16444 is a characteristic of human tumors in vivo , human tumor cell lines , and normal somatic tissue in some individuals . It was hypothesized that P16444 is a risk factor for developing breast cancer in BRCA ( P38398 and P51587 ) germline mutation carriers . To test the hypothesis , human peripheral blood lymphocytes of BRCA carriers with and without breast cancer and healthy non-carrier relatives ( controls ) were cultured for 9 days in medium containing either 0.1 mmol/L L-methionine or 0.2 mmol/L D,L-homocysteine , with the ratio of viable cell growth in both types of medium after 9 days used to calculate the methionine-dependence index ( MDI ) , a measure of P16444 . We also tested whether P16444 was associated with common polymorphisms in methionine metabolism . Viable cell growth , MDI , and polymorphism frequency in Q9UBK8 ( A66G and C524T ) and P42898 ( A1298C and A1793G ) did not differ among the study groups ; however , MDI tended to be higher in BRCA carriers with breast cancer than those without and was significantly increased in P42898 677T allele carriers relative to wild-type carriers ( P=0.017 ) . The presence of Q99707 A2756G mutant allele and P42898 C677T mutant allele in carriers was associated with increased breast cancer risk [ odds ration , 3.2 ( P=0.16 ; 95 % confidence interval , 0.76-13.9 ) and 3.9 ( P=0.09 ; 95 % confidence interval , 0.93-16.3 ) , respectively ] . The results of this study support the hypothesis that defects in methionine metabolism may be associated with breast cancer risk in BRCA carriers . Molecular networks involved in the immune control of BK polyomavirus . BK polyomavirus infection is the important cause of virus-related nephropathy following kidney transplantation . BK virus reactivates in 30 % -80 % of kidney transplant recipients resulting in BK virus-related nephropathy in 1 % -10 % of cases . Currently , the molecular processes associated with asymptomatic infections in transplant patients infected with BK virus remain unclear . In this study we evaluate intrarenal molecular processes during different stages of BKV infection . The gene expression profiles of 90 target genes known to be associated with immune response were evaluated in kidney graft biopsy material using TaqMan low density array . Three patient groups were examined : control patients with no evidence of BK virus reactivation ( n = 11 ) , infected asymptomatic patients ( n = 9 ) , and patients with BK virus nephropathy ( n = 10 ) . Analysis of biopsies from asymptomatic viruria patients resulted in the identification of 5 differentially expressed genes ( P07766 , P34810 , P41597 , P05362 , and P12755 ) ( P < 0.05 ) , and functional analysis showed a significantly heightened presence of costimulatory signals ( e.g. , P25942 / P29965 ; P < 0.05 ) . Gene ontology analysis revealed several biological networks associated with BKV immune control in comparison to the control group . This study demonstrated that asymptomatic BK viruria is associated with a different intrarenal regulation of several genes implicating in antiviral immune response . P25021 mediated relaxation of buffalo ( Bubalus bubalus ) ureter . On the buffalo ureter , histamine did not elicit any direct effect . However , it caused concentration-dependent relaxation of the tissues precontracted by carbachol , phenylephrine , norepinephrine , KCI or BaCl2 and also inhibited the contractile effect of carbachol . DB08805 selectively antagonised the relaxation and inhibition of contractile response but mepyramine did not show this effect . Isoprenaline , dobutamine , salbutamol , verapamil and papaverine neither produced any direct effect nor relaxed the carbachol-contracted tissues ; norepinephrine and epinephrine had contractile effects . Hence , the histamine-induced relaxation was mediated through the activation of H2 receptors and not through adrenergic mechanisms or blockade of Ca(2+)-channels or inhibition of cyclic nucleotide phosphodiesterase .

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interacts_with DB00758?

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Direct and irreversible inhibition of cyclooxygenase-1 by nitroaspirin ( DB05822 ) . Benzoic acid , 2-(acetyl-oxy)-3-[(nitrooxy)methyl]phenyl ester ( DB05822 ) , a new drug made by an aspirin molecule linked , through a spacer , to a nitric oxide ( NO ) -donating moiety , is now under clinical testing for the treatment of atherothrombotic conditions . DB00945 exerts its antithrombotic activity by irreversibly inactivating platelet cyclooxygenase ( P36551 ) -1 . DB05822 in vivo undergoes metabolism into deacetylated and/or denitrated metabolites , and it is not known whether DB05822 needs to liberate aspirin to inhibit P23219 , or whether it can block it as a whole molecule . The aim of our study was to evaluate the effects of DB05822 and its analog or metabolites on platelet P23219 and whole blood P35354 and on purified ovine P36551 ( oCOX ) -1 and oCOX-2 . In particular , we have compared the mechanism by which DB05822 inhibits purified oCOX enzymes with that of aspirin using a spectrophotometric assay . All the DB05822 derivatives containing acetylsalicylic acid inhibited the activity of oCOX-1 and oCOX-2 , whereas the deacetylated metabolites and the nitric oxide-donating moiety were inactive . Dialysis experiments showed that oCOX-1 inhibition by DB05822 , similar to aspirin , is irreversible . Reversible P36551 inhibitors ( indomethacin ) or salicylic acid incubated with the enzyme before DB05822 prevent the irreversible inhibition of oCOX-1 by DB05822 as well as by aspirin . In conclusion , our data show that DB05822 acts as a direct and irreversible inhibitor of P23219 and that the presence of a spacer and NO-donating moiety in the molecule slows the kinetics of P23219 inhibition by DB05822 , compared with aspirin . Wheat germ agglutinin behaves as a DB00644 antagonist but induces gonadotrope desensitization . Preincubation of cultured rat pituitary cells with 10 micrograms/ml of either wheat germ agglutinin ( WGA ) or concanavalin A inhibited LH release stimulated with DB00644 ( 0.5 nM ) by 55 % and 40 % , respectively . WGA-inhibition of LH release stimulated by DB00644 was dose-dependent , reaching a plateau of 75 % inhibition at 50 micrograms/ml . Concomitantly , WGA induced a dose-dependent inhibition of 125I- DB06719 specific binding to pituitary cells , with a maximal inhibition of 45 % . The inhibition of 125I- DB06719 binding by WGA is due to P30968 internalization and not to persistent occupancy of the receptors . In addition to the effect of WGA on receptor internalization , WGA also induced partial desensitization of pituitary cells but was ineffective in modulating DB00644 -induced desensitization . These findings indicate that WGA has all the characteristics of a DB00644 antagonist , nevertheless , it does induce desensitization of cultured rat pituitary cells to further stimulation with DB00644 . DB00233 inhibits O15111 alpha phosphorylation of P25963 in mouse intestinal epithelial cells . P01375 alpha ( TNFalpha ) -stimulated nuclear factor ( NF ) kappaB activation plays a key role in the pathogenesis of inflammatory bowel disease ( Q9UKU7 ) . Phosphorylation of NFkappaB inhibitory protein ( IkappaB ) leading to its degradation and NFkappaB activation , is regulated by the multimeric O15111 complex , including IKKalpha and IKKbeta . We recently reported that DB00244 ( DB00244 ) inhibits TNFalpha-regulated IkappaB degradation and NFkappaB activation . To determine the mechanism of DB00244 inhibition of IkappaB degradation , we studied young adult mouse colon ( YAMC ) cells by immunodetection and in vitro kinase assays . We show DB00244 inhibits TNFalpha-stimulated phosphorylation of P25963 in intact YAMC cells . Phosphorylation of a glutathione S-transferase- P25963 fusion protein by cellular extracts or immunoprecipitated IKKalpha isolated from cells treated with TNFalpha is inhibited by DB00244 . Recombinant IKKalpha and IKKbeta autophosphorylation and their phosphorylation of glutathione S-transferase- P25963 are inhibited by DB00244 . However , IKKalpha serine phosphorylation by its upstream kinase in either intact cells or cellular extracts is not blocked by DB00244 . Surprisingly , immunodepletion of cellular extracts suggests IKKalpha is predominantly responsible for P25963 phosphorylation in intestinal epithelial cells . In summary , DB00244 inhibits TNFalpha-stimulated IKKalpha kinase activity toward P25963 in intestinal epithelial cells . These findings suggest a novel role for DB00244 in the management of Q9UKU7 by disrupting TNFalpha activation of NFkappaB . [ Anticoagulants of primary haemostasis ] . Inhibition of platelet function plays an important role in the treatment and secondary prevention of cardiovascular or cerebrovascular ischemic diseases . Established antiplatelet agents use different pharmacological targets for this role . Acetyl salicylic acid achieves a reduction of thromboxane A2 formation by inhibition of P23219 . DB00208 or clopidogrel are ADP- Q9H244 receptor antagonists . Tirofiban , abciximab or eptifibatid are used for the inhibition of the glycoprotein IIb/IIIa receptor which is activated at the surface of platelets preceding the final step of their aggregation . The mechanism of dipyridamole is based on the inhibition of adenosine uptake and of phosphodiesterase-5 . Efforts are made to improve antiplatelet therapy with the aim to find agents with favorable clinical outcome and lower bleeding risk . Current clinical studies focus on a new generation of ADP receptor antagonists ( prasugrel , cangrelor and ticagrelor ) as successors of ticlopidine and clopidogrel after coronary arterial interventions . Developments using platelet targets different from established drugs are thrombin receptor antagonists ( like SCH530348 ) or thromboxane receptor antagonists ( like S18886/terutroban ) in patients with cerebrovascular events . Results from recent experimental studies could lead to new strategies for antiplatelet therapy ( like inhibition of GP Ib receptor , GP VI receptor , platelet-leukocyte interaction , factor XII and others ) in the future . High loading dose of clopidogrel is unable to satisfactorily inhibit platelet reactivity in patients with glycoprotein IIIA gene polymorphism : a genetic substudy of PRAGUE-8 trial . The study aimed to assess the impact of nine polymorphisms of genes encoding platelet receptors , enzymes , and hemostatic factors on clopidogrel efficacy to inhibit platelet reactivity in patients with stable coronary artery disease undergoing elective coronary angiography either with or without ad hoc percutaneous coronary intervention . The study was performed as a genetic substudy of the PRAGUE-8 trial . Ninety-five patients pretreated with 600 mg clopidogrel at least 6 h prior to coronary angiography were tested . Baseline platelet reactivity to ADP was assessed before the drug was administered . DB00758 efficacy was tested again at 12 and 28 h after administration . Polymorphisms of platelet receptors , glycoprotein ( GP ) Ia ( 807C/T ) , Q9HCN6 ( 13254C/T ) , P05106 ( PlA1/PlA2 ) , P25116 ( IVSn-14A/T ) , Q9H244 ( 32C/T ) , Q9H244 ( H1/H2 ) haplotype , gene variations of cyclooxygenase-1 , Leiden , and factor II mutations were studied . Flow cytometric tests of vasodilator-stimulated phosphoprotein phosphorylation states were used as a measure of drug efficacy . None of the gene polymorphisms influenced baseline ADP-induced platelet reactivity significantly . Twenty-eight hours after drug administration , differences in suppression of ADP-induced platelet reactivity were observed between polymorphism-positive and polymorphism-negative patients . Inhibition of platelet reactivity , after 600 mg of clopidogrel , was significantly less in carriers of PlA2 ( P=0.009 ) for mean decrease in platelet reactivity index . The proportion of clopidogrel nonresponders ( platelet reactivity index > 50 % ) was apparently higher in PlA2 carriers in comparison with PlA1/PlA1 patients ( 54 vs. 24 % , P=0.082 ) . A 600 mg loading dose of clopidogrel failed to acceptably inhibit platelet reactivity in patients who were positive for the PlA2 polymorphism . DB08816 increases adenosine plasma concentration in patients with an acute coronary syndrome . OBJECTIVES : This study aimed to investigate the impact of ticagrelor on adenosine plasma concentration ( P25054 ) in acute coronary syndrome ( ACS ) patients . BACKGROUND : DB08816 is a direct-acting Q9H244 -adenosine diphosphate receptor blocker . The clinical benefit of ticagrelor compared with clopidogrel in ACS patients suggests that the drug has non-platelet-directed properties . Animal and in vitro models suggested that the " pleiotropic " properties of ticagrelor may be related to an interaction with adenosine metabolism . METHODS : We prospectively randomized 60 ACS patients to receive ticagrelor or clopidogrel . The P25054 was measured by liquid chromatography . To assess the mechanism of P25054 variation , we measured adenosine deaminase concentration , adenosine uptake by red blood cells , and cyclic adenosine monophosphate production by cells overexpressing adenosine receptors . The Q9H244 -adenosine diphosphate receptor blockade was assessed by the vasodilator-stimulated phosphoprotein index . RESULTS : Patients receiving ticagrelor had significantly higher P25054 than patients receiving clopidogrel ( 1.5 μM [ interquartile range : 0.98 to 1.7 μM ] vs. 0.68 μM [ interquartile range : 0.49 to 0.78 μM ] ; p < 0.01 ) . The P25054 was not correlated with vasodilator-stimulated phosphoprotein ( p = 0.16 ) . Serum-containing ticagrelor inhibited adenosine uptake by red blood cells compared with clopidogrel or controls ( p < 0.01 for both comparisons ) . DB00640 deaminase activity was similar in serum of patients receiving clopidogrel or ticagrelor ( p = 0.1 ) . DB08816 and clopidogrel had no direct impact on adenosine receptors ( p = not significant ) . CONCLUSIONS : DB08816 increases P25054 in ACS patients compared with clopidogrel by inhibiting adenosine uptake by red blood cells . DB00055 mediates a healing phenotype in cultured tenocytes . Tendon injuries cause considerable morbidity in the general adult population . The tenocytes within the tendon have the full capacity to heal the tendon intrinsically . DB00055 ( P25054 ) plays an important role in coagulation and inflammation and more recently has been shown to promote cutaneous wound healing . In this study we examined whether P25054 can induce a wound healing phenotype in tenocytes . Sheep tenocytes were treated with P25054 , endothelial protein C receptor ( Q9UNN8 ) blocking antibody ( RCR252 ) and/or Q9UNN8 small interfering (si)RNA . Cell proliferation and migration were measured by crystal violet assay and a scratch wounding assay , respectively . The expression of Q9UNN8 , matrix metalloproteinase ( MMP ) -2 , type I collagen and Q96HU1 kinase activity were detected by real time PCR , zymography , immunofluorescence , immunohistochemistry and Western blotting . P25054 stimulated proliferation , P08253 activity and type I collagen deposition in a dose-dependent manner and promoted migration of cultured tenocytes . P25054 dose-dependently stimulated phosphorylated ( P ) - P28482 and inhibited P-p38 . Interestingly , tenocytes expressed Q9UNN8 protein , which was up-regulated by P25054 . When tenocytes were pre-treated with RCR252 or Q9UNN8 siRNA the effect of P25054 on proliferation , P08253 and type 1 collagen synthesis and Q96HU1 kinases was blocked . P25054 promotes the growth , P08253 activity , type I collagen deposition and migration of tenocytes . Furthermore , Q9UNN8 is expressed by tenocytes and mediates the actions of P25054 , at least partly by signalling through selective Q96HU1 kinases . These data implicate P25054 as a potential healing agent for injured tendons . DB04849 : a highly potent , orally bioavailable , vascular endothelial growth factor receptor-2 tyrosine kinase inhibitor for the treatment of cancer . Inhibition of vascular endothelial growth factor-A ( P15692 ) signaling is a promising therapeutic approach that aims to stabilize the progression of solid malignancies by abrogating tumor-induced angiogenesis . This may be accomplished by inhibiting the kinase activity of P15692 receptor-2 ( P35968 ) , which has a key role in mediating P15692 -induced responses . The novel indole-ether quinazoline DB04849 is a highly potent ( IC50 < 1 nmol/L ) DB00171 -competitive inhibitor of recombinant P35968 tyrosine kinase in vitro . Concordant with this activity , in human umbilical vein endothelial cells , DB04849 inhibited P15692 -stimulated proliferation and P35968 phosphorylation with IC50 values of 0.4 and 0.5 nmol/L , respectively . In a fibroblast/endothelial cell coculture model of vessel sprouting , DB04849 also reduced vessel area , length , and branching at subnanomolar concentrations . Once-daily oral administration of DB04849 ablated experimental ( P15692 -induced ) angiogenesis in vivo and inhibited endochondral ossification in bone or corpora luteal development in ovary ; physiologic processes that are highly dependent upon neovascularization . The growth of established human tumor xenografts ( colon , lung , prostate , breast , and ovary ) in athymic mice was inhibited dose-dependently by DB04849 , with chronic administration of 1.5 mg per kg per day producing statistically significant inhibition in all models . A histologic analysis of Calu-6 lung tumors treated with DB04849 revealed a reduction in microvessel density within 52 hours that became progressively greater with the duration of treatment . These changes are indicative of vascular regression within tumors . Collectively , the data obtained with DB04849 are consistent with potent inhibition of P15692 signaling , angiogenesis , neovascular survival , and tumor growth . DB04849 is being developed clinically as a once-daily oral therapy for the treatment of cancer . Structure and function of eritadenine and its 3-deaza analogues : potent inhibitors of S-adenosylhomocysteine hydrolase and hypocholesterolemic agents . d- DB03769 ( DEA ) is a potent inhibitor of S-adenosyl-l-homocysteine hydrolase ( P23526 ) and has hypocholesterolemic activity . We have hypothesized that 3-deaza-DEA ( P01024 -DEA ) and its analogues retain high level of P23526 inhibitory activity and have resistance to deamination and glycosidic bond hydrolysis in vivo . Such P01024 -DEA analogues would have much higher hypocholesterolemic activity . P01024 -DEA , and its methyl ester ( P01024 -OMeDEA ) and its methyl amido ( P01024 -NMeDEA ) were synthesized to examine their P23526 inhibitory and hypocholesterolemic activities . A crystal structure of P23526 containing P01024 -DEA was determined and confirmed that DEA and P01024 -DEA bound to the same site of P23526 with the same binding mode . The P23526 inhibitory activities of P01024 -DEA ( K(I)=1.5 microM ) and P01024 -OMeDEA ( K(I)=1.5 microM ) are significantly lower than that of DEA ( K(I)=30 nM ) , while rats fed by P01024 -DEA and P01024 -OMeDEA decrease the total plasma cholesterol and phospholipids by 36-40 % and 23 % , respectively , which is similar to the level of reductions ( 42 % and 27 % ) by DEA . P01024 -NMeDEA lost most of the P23526 inhibitory activity ( K(I)=30 microM ) and dietary P01024 -NMeDEA does not decrease cholesterol and phospholipid in plasma but decreases the triacylglycerol level by 16 % . DEA and P01024 -DEA analogues are neither substrates nor inhibitors of adenosine deaminase . Comparison of three GPCR structural templates for modeling of the Q9H244 nucleotide receptor . The P2Y(12) receptor ( P2Y(12)R ) is an ADP-activated G protein-coupled receptor ( GPCR ) that is an important target for antithrombotic drugs . Three homology models of P2Y(12)R were compared , based on different GPCR structural templates : bovine rhodopsin ( bRHO ) , human A(2A) adenosine receptor ( A(2A)AR ) , and human P61073 ( P61073 ) . By criteria of sequence analysis ( 25.6 % identity in transmembrane region ) , deviation from helicity in the second transmembrane helix ( TM2 ) , docked poses of ligands highlighting the role of key residues , accessibility of a conserved disulfide bridge that is reactive toward irreversibly-binding antagonists , and the presence of a shared disulfide bridge between the third extracellular loop ( EL3 ) and the N-terminus , the P61073 -based model appeared to be the most consistent with known characteristics of P2Y(12)R . The docked poses of agonist 2MeSADP and charged anthraquinone antagonist PSB-0739 in the binding pocket of P2Y(12)R-CXC agree with previously published site-directed mutagenesis studies of Arg256 and Lys280 . A sulfonate at position 2 of the anthraquinone core created a strong interaction with the Lys174(EL2) side chain . The docking poses of the irreversibly-binding , active metabolite ( existing as two diastereoisomers in vivo ) of the clinically utilized antagonist DB00758 were compared . The free thiol group of the 4S diastereoisomer , but not the 4R isomer , was found in close proximity ( ~4.7 Å ) to the sulfur atom of a disulfide bridge involving Cys175 , suggesting greater activity in covalent binding . Therefore , ligand docking to the P61073 -based model of the P2Y(12)R predicted poses of both reversibly and irreversibly-binding small molecules , consistent with observed pharmacology and mutagenesis studies . The formation of monocyte-platelet aggregates is independent of on-treatment residual agonists'-inducible platelet reactivity . BACKGROUND : Circulating monocyte-platelet aggregates ( DB00603 ) are a sensitive marker of in vivo platelet activation and patients with atherosclerotic vascular disease exhibit higher levels of DB00603 . DB00758 has been shown to reduce DB00603 formation in these patients to a greater extent than aspirin . However , response to clopidogrel and aspirin shows a wide variability , and patients with high on-treatment residual platelet reactivity are at an increased risk for adverse events after coronary stenting . We therefore investigated the association of DB00603 with on-treatment residual agonists'-inducible platelet aggregation in 125 patients on dual antiplatelet therapy after peripheral , coronary or carotid artery stenting . METHODS : DB00603 were characterized by co-expression of monocyte marker P08571 and platelet-specific markers ( CD42b and CD62P ) by whole blood flow cytometry . Platelet reactivity was determined by light transmission aggregometry , the VerifyNow Q9H244 and aspirin assays , and the vasodilator-stimulated phosphoprotein phosphorylation assay . Cut-off values for residual platelet reactivity were defined according to quartiles of each assay . RESULTS : The extent of DB00603 formation showed no significant differences between patients without and with residual ADP-inducible platelet reactivity , and between individuals without and with residual arachidonic acid ( AA ) -inducible platelet reactivity . Even patients with combined on-treatment residual ADP- and AA-inducible platelet reactivity did not exhibit significantly higher levels of DB00603 than patients without any on-treatment residual platelet reactivity . CONCLUSION : High on-treatment residual agonists'-inducible platelet reactivity results in less than a 25 % increase in circulating DB00603 , suggesting that DB00603 formation is largely dependent on other factors . Characterization of the aggregation responses of camel platelets . BACKGROUND : Despite evidence of active hemostasis , camel platelets barely respond to common aggregating agents at standard doses used for human platelet aggregation . OBJECTIVES : The purpose of the study was to find out whether camel platelets can be activated by high doses or combinations of aggregation agonists , and to characterize the receptor that mediates the aggregation response to adenosine diphosphate ( ADP ) , the most potent agonist for camel platelets known so far . METHODS : Aggregation studies were performed with platelet-rich plasma ( PRP ) in response to multiple doses or combinations of ADP , epinephrine ( P08473 ) , collagen , and arachidonic acid ( AA ) . Aggregation responses to ADP were performed before and after the addition of the ADP receptor ( Q9H244 ) antagonist DB00758 . RESULTS : Camel platelets responded to ADP at doses higher than the standard dose for human platelets , and to combinations of P08473 and other agonists , while no aggregation was elicited with P08473 or AA alone . DB00758 blocked the ADP-induced aggregation responses in a dose-dependent fashion in vitro . CONCLUSIONS : Camel platelet aggregation can be activated by increasing the dose of some agonists such as ADP , but not AA or P08473 . Irreversible aggregation of camel platelets could also be triggered by a combination of P08473 and ADP , and collagen and AA . Inhibition with clopidogrel suggests that camel platelets express the ADP receptor , Q9H244 . Understanding platelet function in camels will add to the understanding of platelet function in health and disease . Stereospecific interaction of a novel spirosuccinimide type aldose reductase inhibitor , DB05327 , with aldose reductase . P15121 ( AR ) is an NADPH-dependent enzyme implicated in diabetic complications . DB05327 [ ( R ) -(-)-2-(4-bromo-2-fluorobenzyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-4-spiro-3'-pyrrolidine-1,2',3,5'-tetrone ] is a structurally novel and potent Q9Y4X5 with an inhibitor constant ( K(i) = 10(-)(10) M ) 2000-fold lower than that of its optical antipode ( S-isomer ) . To elucidate the inhibition modes and the stereochemical differences in their inhibitory potencies , we examined the interaction of these R- and S-isomers with AR under physiological conditions . Enzyme kinetic analysis , which was performed by using physiological substrates at 37 degrees C , showed that both isomers selectively act on the E-NADP(+) complex in both the forward and reverse reactions of AR . However , fluorometric titration analysis demonstrated that the affinities of the isomers for the E-NADP(+) complex are about the same as those for the E-NADPH complex and the apoenzyme . These results suggested that the selective binding to the E-NADP(+) complex arises from the predominance of this enzyme form during steady-state turnover rather than from binding specificity . Both the competition with a known active site-directed Q9Y4X5 and the protective effect on AR inactivation by N-bromosuccinimide showed that the isomers bind to the active site of the enzyme , but the thermodynamic parameters for the binding to AR indicated that additional hydrogen bonds and/or van der Waals interactions contribute to the energetic stabilization in the E-R-isomer complex . Molecular modeling , together with the deductions from spectroscopic studies , suggested that the succinimide ring and the 4-bromo-2-fluorobenzyl group of the R-isomer are optimally located for formation of a hydrogen-bonding network with AR , and that the latter benzyl group is also effective for the differentiation between AR and aldehyde reductase ( a closely related enzyme ) . A case of tuberculous arthritis following the use of etanercept . DB00005 is a tumor necrosis factor ( P01375 ) inhibitor that has been used for the treatment of chronic inflammatory diseases including rheumatoid arthritis , ankylosing spondylitis and psoriatic arthritis . Because of its immunosuppressive activity , opportunistic infections have been noted in treated patients , most notably caused by Mycobacterium tuberculosis . Tuberculosis may present in an extrapulmonary or disseminated form . Since P01375 inhibitors have been used in Korea , a few cases of P01375 inhibitor associated tuberculosis have been described . However , tuberculous arthritis has not been previously reported . We describe a case of tuberculous arthritis in a 57-year-old woman with rheumatoid arthritis who was treated with etanercept . Stress-induced changes in the expression of monocytic beta2-integrins : the impact of arousal of negative affect and adrenergic responses to the Anger Recall Interview . Adhesion of circulating monocytes to the vascular endothelium is one of the earliest steps in the development of atherosclerosis . This leukocyte-to-endothelium interaction is mediated in part by beta2-integrins , a group of cell adhesion molecules that bind to endothelial ligands . Given the significance of this interaction to atherogenesis , we examined the effects of stress , operationalized as the arousal of negative affect ( NA ) and cardiovascular and catecholamine responses to the Anger Recall Interview ( Q9Y4X5 ) , on the expression of LFA-1 ( CD11a ) , Mac-1 ( CD11b ) and p150/95 ( CD11c ) on circulating monocytes ( P08571 + ) . Subjects were 173 healthy , nonsmoking men and women ( 60 % men , 40 % minorities , aged 18-49 year ) . Arousal of NA , cardiovascular responses ( heart rate [ HR ] , systolic blood pressure [ SBP ] , diastolic blood pressure [ DBP ] ) , circulating catecholamines ( epinephrine [ Epi ] , norepinephrine [ Ne ] ) and beta2-integrin ( CD11/ P05107 ) expression were determined prior to and following the Q9Y4X5 . The principal findings were that the Q9Y4X5 , on average , induced a decrease in monocyte expression of beta2-integrins . However , after adjusting for age , sex , body mass index , exercise status , and baseline level of beta2-integrin expression , those individuals who showed the largest increases in NA , Ne and DBP during the Q9Y4X5 showed an increase in monocyte beta2-integrin expression . Thus , heightened psychological and physiological stress responses induced phenotypic changes in monocytic expression of beta2-integrins that are consistent with the role of monocytes/macrophages in vascular inflammation and increased risk of atherosclerotic cardiovascular disease . Mechanisms of aspirin resistance . DB00945 is integral to the secondary prevention of cardiovascular disease and acts to impair the development of platelet-mediated atherothromboembolic events by irreversible inhibition of platelet cyclooxygenase-1 ( P23219 ) . Inhibition of this enzyme prevents the synthesis of the potent pro-aggregatory prostanoid thromboxane A2 . A large number of patients continue to experience atherothromboembolic events despite aspirin therapy , so-called ' aspirin treatment failure ' , and this is multifactorial in aetiology . Approximately 10 % however do not respond appropriately to aspirin in a phenomenon known as ' aspirin resistance ' , which is defined by various laboratory techniques . In this review we discuss the reasons for aspirin resistance in a systematic manner , starting from prescription of the drug and ending at the level of the platelet . Poor medication adherence has been shown to be a cause of apparent aspirin resistance , and may in fact be the largest contributory factor . Also important is high platelet turnover due to underlying inflammatory processes , such as atherosclerosis and its complications , leading to faster regeneration of platelets , and hence of P23219 , at a rate that diminishes the efficacy of once daily dosing . Recent developments include the identification of platelet glycoprotein IIIa as a potential biomarker ( as well as possible underlying mechanism ) for aspirin resistance and the discovery of an anion efflux pump that expels intracellular aspirin from platelets . The absolute as well as relative contributions of such factors to the phenomenon of aspirin resistance are the subject of continuing research . Intracellular signaling pathways activated by kisspeptins through Q969F8 : do multiple signals underlie function diversity ? Kisspeptins , a family of peptide products derived from the KiSS-1 gene , activate their cognate receptor Q969F8 in various target tissues to exert disparate functions , including inhibition of tumor metastasis and control of reproductive function . In contrast to the plethora of studies that have analyzed in recent years the regulatory functions of the KiSS-1/ Q969F8 system , only a limited number of reports have been primarily focused on delineating the intracellular signaling pathways involved . Nevertheless , there is solid evidence indicating that kisspeptin can activate a wide variety of signals via Q969F8 . These include typical G-protein ( Galphaq/11 ) -coupled cascades , such as activation of phospholipase C ( P98160 ) , and subsequent accumulation of inositol-(1,4,5)-triphosphate ( IP3 ) , intracellular Ca(2+) mobilization , and activation of protein kinase C . However , kisspeptin also activates pathways related to mitogen activated protein kinases ( MAPK ) , especially P27361 /2 , and p38 and phosphatidylinositol-3-kinase ( PI3K ) /Akt . Additionally , the kisspeptin/ Q969F8 pair can also influence cell signaling by interacting with other receptors , such as chemokine receptor P61073 , and P30968 . Kisspeptin can also affect other signaling events , like expression of matrix metalloproteinase 9 ( via NFkappaB ) , and that of calcineurin . The information gathered hitherto clearly indicates that activation of a specific set of interconnected signals is selectively triggered by kisspeptin via Q969F8 in a cell type-dependent manner to precisely regulate functions as distinct as hormone release and cell migration . In this scenario , it will be important to decipher kisspeptin/ Q969F8 signaling mechanisms in reproductive and non-reproductive tissues by studying additional models , especially on natural kisspeptin targets expressing endogenous Q969F8 . Complement activation triggered by chondroitin sulfate released by thrombin receptor-activated platelets . BACKGROUND : DB09301 ( CS ) is a glycosaminoglycan released by activated platelets . OBJECTIVE : Here we test the hypothesis that CS released by activated platelets can trigger complement activation in the fluid phase . METHODS AND RESULTS : P25116 -activating peptide ( TRAP ) -6 was used to activate platelets in platelet-rich plasma and blood , anticoagulated with the thrombin inhibitor lepirudin . TRAP activation induced fluid-phase complement activation , as reflected by the generation of C3a and sC5b-9 , which could be attenuated by the P01024 inhibitor compstatin . P34059 ABC treatment of supernatants from activated platelets totally inhibited the activation , indicating that platelet-derived CS had initiated the complement activation . Furthermore , addition of purified CS to plasma strongly triggered complement activation . C1q was identified as the recognition molecule , as it bound directly to CS , and CS-triggered complement activation could be restored in C1q-depleted serum by adding purified C1q . TRAP activation of whole blood increased the expression of CD11b on leukocytes and generation of leukocyte-platelet complexes . It was demonstrated that these leukocyte functions were dependent on P01024 activation and signaling via C5a , as this expression could be inhibited by compstatin and by a P21730 antagonist . CONCLUSIONS : We conclude that platelets trigger complement activation in the fluid phase by releasing CS , which leads to inflammatory signals mediated by C5a . DB00173 nucleotides inhibit cytokine generation by human mast cells through a Gs-coupled receptor . DB00171 and ADP activate functionally distinct G protein-coupled purinergic ( P2Y ) receptors . We determined the expression and function of adenine nucleotide-specific P2Y receptors on cord blood-derived human mast cells ( hMCs ) . Human MCs expressed mRNA encoding the ADP-specific P47900 , Q9H244 , and Q9BPV8 receptors ; the DB00171 /UTP-specific P41231 receptor ; and the DB00171 -selective Q96G91 receptor . ADP ( 0.05-50 muM ) induced calcium flux that was completely blocked by a P47900 receptor-selective antagonist and was not cross-desensitized by DB00171 . Low doses of ADP induced strong phosphorylation of P29323 and p38 MAPKs ; higher doses stimulated eicosanoid production and exocytosis . Although MAPK phosphorylation was blocked by a combination of P47900 - and Q9H244 -selective antagonists , neither interfered with secretion responses . Unexpectedly , both ADP and DB00171 inhibited the generation of P01375 in response to the O60603 ligand , peptidoglycan , and blocked the production of P01375 , P10145 , and MIP-1beta in response to leukotriene D(4) . These effects were mimicked by two DB00171 analogues , adenosine 5'-O-(3-thiotriphosphate) and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate ( BzATP ) , but not by adenosine . ADP , DB00171 , adenosine 5'-O-(3-thiotriphosphate) , and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate each induced DB02527 accumulation , stimulated the phosphorylation of CREB , and up-regulated the expression of inducible DB02527 early repressor , a CREB-dependent inhibitor of cytokine transcription . Human MCs thus express several ADP-selective P2Y receptors and at least one G(s)-coupled ADP/ DB00171 receptor . Nucleotides could therefore contribute to MC-dependent microvascular leakage in atherosclerosis , tissue injury , and innate immunity while simultaneously limiting the extent of subsequent inflammation by attenuating the generation of inducible cytokines by MCs . Temporal and pharmacological characterization of angiostatin release and generation by human platelets : implications for endothelial cell migration . Platelets play an important role in thrombosis and in neo-vascularisation as they release and produce factors that both promote and suppress angiogenesis . Amongst these factors is the angiogenesis inhibitor angiostatin , which is released during thrombus formation . The impact of anti-thrombotic agents and the kinetics of platelet angiostatin release are unknown . Hence , our objectives were to characterize platelet angiostatin release temporally and pharmacologically and to determine how angiostatin release influences endothelial cell migration , an early stage of angiogenesis . We hypothesized anti-platelet agents would suppress angiostatin release but not generation by platelets . Human platelets were aggregated and temporal angiostatin release was compared to vascular endothelial growth factor ( P15692 ) . Immuno-gold electron microscopy and immunofluorescence microscopy identified α-granules as storage organelles of platelet angiostatin . Acetylsalicylic acid , MRS2395 , P08514 /IIIa blocking peptide , and aprotinin were used to characterize platelet angiostatin release and generation . An endothelial cell migration assay was performed under hypoxic conditions to determine the effects of pharmacological platelet and angiostatin inhibition . Compared to P15692 , angiostatin generation and release from α-granules occurred later temporally during platelet aggregation . Consequently , collagen-activated platelet releasates stimulated endothelial cell migration more potently than maximally-aggregated platelets . Platelet inhibitors prostacyclin , S-nitroso-glutathione , acetylsalicylic acid , and P08514 /IIIa blocking peptide , but not a Q9H244 inhibitor , suppressed angiostatin release but not generation . Suppression of angiostatin generation in the presence of acetylsalicylic acid enhanced platelet-stimulated endothelial migration . Hence , the temporal and pharmacological modulation of platelet angiostatin release may have significant consequences for neo-vascularization following thrombus formation .

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interacts_with DB09048?

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Angiogenic alterations associated with circulating neoplastic DNA in ovarian carcinoma . OBJECTIVES : Forty percent of women with ovarian carcinoma have circulating free neoplastic DNA identified in plasma . Angiogenesis is critical in neoplastic growth and metastasis . We sought to determine whether circulating neoplastic DNA results from alterations in the balance of angiogenesis activators and inhibitors . METHODS : Sixty patients with invasive ovarian carcinomas with somatic P04637 mutations that had been characterized for circulating neoplastic DNA had carcinoma analyzed for microvessel density using immunohistochemistry with CD31 and for the expression of P15692 , Q15389 , O15123 , P35354 , P00749 , P07996 , P09603 , P42336 , Q16665 , P10145 , P08253 , and P14780 message by real-time quantitative polymerase chain reaction . The expression of each gene was calculated relative to P04406 expression for each neoplasm . Patient plasma had been tested for circulating neoplastic DNA using a ligase detection reaction . RESULTS : P08253 expression was significantly correlated with free plasma neoplastic DNA ( P = .007 ) . Microvessel density was not correlated with plasma neoplastic DNA or P38398 /2 mutation status . The expression pattern of other angiogenic factors did not correlate with plasma neoplastic DNA but correlated with each other . P38398 /2 mutated carcinomas had significantly different expression profiles of angiogenesis activators and inhibitors in comparison to sporadic carcinomas . CONCLUSIONS : P08253 expression is associated with the presence of circulating neoplastic DNA in women with ovarian carcinoma . These data are consistent with the proinvasive properties of P08253 and suggest that the presence of circulating neoplastic DNA indicates a more aggressive malignant phenotype . Carcinomas with germ line P38398 /2 mutations had a lower angiogenic profile than those without mutations . P42892 and β-arrestins exert spatiotemporal control of DB05875 -induced inflammatory signals . Although the intracellular trafficking of G protein-coupled receptors controls specific signaling events , it is unclear how the spatiotemporal control of signaling contributes to complex pathophysiological processes such as inflammation . By using bioluminescence resonance energy transfer and superresolution microscopy , we found that DB05875 ( SP ) induces the association of the neurokinin 1 receptor ( P25103 ) with two classes of proteins that regulate SP signaling from plasma and endosomal membranes : the scaffolding proteins β-arrestin ( βARRs ) 1 and 2 and the transmembrane metallopeptidases ECE-1c and ECE-1d . In HEK293 cells and non-transformed human colonocytes , we observed that G protein-coupled receptor kinase 2 and β P49407 /2 terminate plasma membrane Ca(2+) signaling and initiate receptor trafficking to endosomes that is necessary for sustained activation of ERKs in the nucleus . βARRs deliver the SP- P25103 endosomes , where P42892 associates with the complex , degrades SP , and allows the P25103 , freed from βARRs , to recycle . Thus , both P42892 and βARRs mediate the resensitization of P25103 Ca(2+) signaling at the plasma membrane . Sustained exposure of colonocytes to SP activates NF-κB and stimulates P10145 secretion . This proinflammatory signaling is unaffected by inhibition of the endosomal P29323 pathway but is suppressed by P42892 inhibition or β P32121 knockdown . Inhibition of protein phosphatase 2A , which also contributes to sustained P25103 signaling at the plasma membrane , similarly attenuates P10145 secretion . Thus , the primary function of βARRs and P42892 in SP-dependent inflammatory signaling is to promote resensitization , which allows the sustained P25103 signaling from the plasma membrane that drives inflammation . [ Role of neurokinin-1 receptor in lung injury in rats with acute necrotizing pancreatitis ] . OBJECTIVE : To investigate the expression of neurokinin-1 receptor ( P25103 ) in the lung tissue , and the relationship between expression of P25103 and lung injury in rats with acute necrotizing pancreatitis ( P01160 ) . METHODS : One hundred and twenty adult Sprague-Dawley rats were randomly divided into P01160 and control groups . Animals in group P01160 were induced by the retrograde intraductal infusion of 5 % sodium taurocholate ( 0.1 ml/kg ) , and animals in normal control group received laparotomy only . The accumulation of polymorphonuclear leukocytes in lung tissues was measured with myeloperoxidase ( P05164 ) assay . Lung endothelial barrier destruction was measured by lung capillary permeability ( LCP ) . Reverse transcription polymerase chain reaction ( RT-PCR ) was used to determine the mRNA expression of P25103 , western blot analysis was used to determine P25103 protein expression levels , and immunohistochemistry was used to localize expression site of P25103 . RESULTS : P25103 mRNA level was enhanced in the lung of P01160 compared with normal control group . Western blot analysis showed overexpression of P25103 protein level exited in P01160 group . Statistical analysis revealed correlation between P25103 mRNA and P05164 ( r=0.83 , P < 0.01 ) and LCP ( r=0.79 , P < 0.01 ) respectively . With immunohistochemistry staining , moderate to strong P25103 immunoreactivity was localized to alveolar membrane , I epithelium , II epithelium and polymorphonuclear leukocytes in the lung of P01160 . CONCLUSION : In P01160 , overexpression of P25103 contributes to disturbance of neuropeptides loop , resulting in aggregation of neutrophilic granulocyte and promoting deterioration of lung injury . The role of the spinal opioid receptor like1 receptor , the P25103 , and cyclooxygenase-2 in maintaining postoperative pain in the rat . Postoperative incident pain is not easily treated with opioids . Mechanical hyperalgesia induced by skin incision in rats is one of the animal models of postoperative incident pain . It is thought that mechanical hyperalgesia is maintained by the sensitization of spinal dorsal horn neurons . The P25103 , the opioid receptor like1 ( P41146 ) receptor , and cyclooxygenase ( P36551 ) -2 reportedly are involved in the development of spinal sensitization . In this study , we clarified the role of the P25103 , the P41146 receptor , and P35354 in the maintenance of mechanical hyperalgesia induced by skin incision . A 1-cm longitudinal incision was made through skin and fascia of the plantar aspect of the right foot in the rat . Four hours after the skin incision , significant mechanical hyperalgesia developed . An P41146 receptor agonist ( nociceptin ) , P25103 antagonists ( CP-96,345 and FK888 ) , and P35354 inhibitors ( NS398 and JTE522 ) were administered intrathecally 4 h after the skin incision . An P41146 receptor agonist and P25103 antagonists , but not P35354 inhibitors , significantly attenuated the level of mechanical hyperalgesia induced by the skin incision . These findings suggest that the spinal P41146 receptor and the P25103 play an important role in maintaining the mechanical hyperalgesia induced by skin incision . IMPLICATIONS : Intrathecal injection of an P25103 antagonist and an P41146 receptor agonist may be effective for the treatment of postoperative incident pain . Hepatic DB00171 -binding cassette transporter A1 is a key molecule in high-density lipoprotein cholesteryl ester metabolism in mice . OBJECTIVE : Mutations in DB00171 -binding cassette transporter A1 ( O95477 ) , the cellular lipid transport molecule mutated in Tangier disease , result in the rapid turnover of high-density lipoprotein ( HDL ) -associated apolipoproteins that presumably are cleared by the kidneys . However , the role of O95477 in the liver for HDL apolipoprotein and cholesteryl ester ( CE ) catabolism in vivo is unknown . METHODS AND RESULTS : Murine HDL was radiolabeled with 125I in its apolipoprotein and with [3H]cholesteryl oleyl ether in its CE moiety . HDL protein and lipid metabolism in plasma and HDL uptake by tissues were investigated in O95477 -overexpressing bacterial artificial chromosome ( BAC ) -transgenic ( BAC+ ) mice and in mice harboring deletions of total ( O95477 -/- ) and liver-specific O95477 ( O95477 (-L/-L) ) . In BAC+ mice with elevated O95477 expression , fractional catabolic rates ( FCRs ) of both the protein and the lipid tracer were significantly decreased in plasma and in the liver , yielding a diminished hepatic selective CE uptake from HDL . In contrast , O95477 -/- or O95477 (-L/-L) mice had significantly increased plasma and liver FCRs for both HDL tracers . An O95477 deficiency was associated with increased selective HDL CE uptake by the liver under all experimental conditions . CONCLUSIONS : Hepatic O95477 has a critical role for HDL catabolism in plasma and for HDL selective CE uptake by the liver . Efficacy and safety of repeated dosing of netupitant , a neurokinin-1 receptor antagonist , in treating overactive bladder . AIM : NK-1 receptors in sensory nerves , the spinal cord and bladder smooth muscle participate in complex sensory mechanisms that regulate bladder activity . This study was designed to assess the efficacy and safety of a new P25103 antagonist , netupitant , in patients with OAB . METHODS : This was a phase II , multicenter , double-blind study in which adults with OAB symptoms > 6 months were randomized to receive 1 of 3 doses of netupitant ( 50 , 100 , 200 mg ) or placebo once daily for 8 weeks . The primary efficacy endpoint was percentage change from baseline in average number of daily micturitions at week 8 . Urinary incontinence , urge urinary incontinence ( UUI ) , and urgency episodes were also assessed . RESULTS : The primary efficacy endpoint was similar in the treatment groups ( -13.85 for placebo to -16.17 in the netupitant 200 mg group ) with no statistically significant differences between netupitant and placebo . The same was true for most secondary endpoints although a significant difference for improvement in UUI episodes and a trend for the greatest decrease in urgency episodes were seen in the netupitant 100 mg group . DB09048 was well tolerated with most treatment emergent adverse events ( AEs ) being mild . While the overall incidence of AEs increased with netupitant dose , there was no evidence for this dose dependency based on relationship to treatment , intensity , or time to onset . CONCLUSIONS : The study failed to demonstrate superiority of netupitant versus placebo in decreasing OAB symptoms , despite a trend favoring netupitant 100 mg . There were no safety concerns with daily administration of netupitant over 8 weeks . Q9BYW2 -α and survivin involved in the anti-apoptotic effect of DB02342 after global ischemia in rats . Survivin is an anti-apoptotic gene that decreases the apoptosis by depressing the expression of caspase-3 . Hypoxia-inducible factor-1-alpha ( Q16665 ) is a transcription factor specifically activated by hypoxia . 2-methoxyestradiol ( DB02342 ) is an estradiol derivative and a known Q16665 inhibitor . DB02342 decreased apoptosis by inhibiting Q16665 . The aim of the present study was to investigate if survivin is involved in the anti-apoptotic effect of DB02342 . Male adult rats were used to make the global ischemia ( GI ) model . Ten minutes after GI , DB02342 was injected intraperitoneally ( 16 mg/kg weight ) . Rats were killed at 6 hours , 12 hours , 24 hours , 48 hours , 96 hours , and 7 days . GI produced a marked increase in Q16665 expressions in the hippocampus at 6 hours and peaked at 48-96 hours . The expressions of survivin and caspase-3 were increased lightly in a similar time course . These molecular changes were accompanied by massive cell loss and apoptosis in the hippocampal regions . DB02342 treatment reduced the expression of Q16665 , increased survivin expression , and decreased the expression of caspase-3 . These results indicate that survivin and Q16665 were involved in the anti-apoptotic effect of DB02342 treated following GI . DB02342 may decrease the Q16665 expression , up-regulate the survivin expression , inhibit the expression of caspase-3 , and finally reduce apoptosis after GI . Synteny mapping of five human chromosome 7 genes on bovine chromosomes 4 and 21 . Five genes on human chromosome 7 ( HSA 7 ) were assigned to bovine chromosome 21 ( BTA 21 ) and 4 ( BTA 4 ) using a bovine-rodent somatic hybrid cell panel . These five genes were alpha-I subunit of adenylate cyclase-inhibiting G-protein ( P63096 ) , alpha/beta preprotachykinin ( P20366 ) , reelin ( P78509 ) , c-AMP dependant protein kinase type II beta regulatory chain ( P31323 ) and apolipoprotein A1 regulatory protein 1 ( P24468 ) . Four genes mapped to BTA 4 ( P63096 , P20366 , P78509 , P31323 ) while one gene mapped to BTA 21 ( P24468 ) . This study confirms the synteny conservation between HSA 7 and BTA 4 , finely maps the breakpoints of conserved synteny on HSA 7 and defines a new synteny conservation between HSA 7 and BTA 21 . Prenatal exposure to bisphenol A promotes angiogenesis and alters steroid-mediated responses in the mammary glands of cycling rats . Prenatal exposure to Q03001 disturbs mammary gland histoarchitecture and increases the carcinogenic susceptibility to chemical challenges administered long after Q03001 exposure . Our aim was to assess the effect of prenatal Q03001 exposure on mammary gland angiogenesis and steroid hormone pathways in virgin cycling rats . Pregnant Wistar rats were exposed to either 25 or 250 g/kg/day ( 25 and 250 Q03001 , respectively ) or to vehicle . Female offspring were autopsied on postnatal day ( P01160 ) 50 or 110 . Ovarian steroid serum levels , the expression of steroid receptors and their co-regulators Q9Y6Q9 and Q9Y618 in the mammary gland , and angiogenesis were evaluated . At P01160 50 , all Q03001 -treated animals had lower serum levels of progesterone , while estradiol levels remained unchanged . The higher dose of Q03001 increased mammary ERα and decreased Q9Y6Q9 expression at P01160 50 and P01160 110 . Q9Y618 protein levels were similar among groups at P01160 50 , whereas at P01160 110 , animals exposed to 250 Q03001 showed a lower Q9Y618 expression . Interestingly , in the control and 25 Q03001 groups , Q9Y618 increased from P01160 50 to P01160 110 . At P01160 50 , an increased vascular area associated with higher P15692 expression was observed in the 250 Q03001 -treated rats . At P01160 110 , the vascular area was still increased , but P15692 expression was similar to that of control rats . The present results demonstrate that prenatal exposure to Q03001 alters the endocrine environment of the mammary gland and its angiogenic process . Increased angiogenesis and altered steroid hormone signals could explain the higher frequency of pre-neoplastic lesions found later in life . This article is part of a Special Issue entitled ' Endocrine disruptors ' . P19957 kinase/AKT pathway as a therapeutic target in multiple myeloma . The development of novel therapies for multiple myeloma depends on a comprehensive understanding of the events leading to cellular proliferation and survival . Controlling pathways that regulate growth signals is an emerging and complementary approach to myeloma treatment . The PI3K/Akt pathway is a central gatekeeper for crucial cellular functions including adhesion , angiogenesis , migration and development of drug resistance . Established proteins and genes such as P42345 , p53 , NF-kappaB and Q92934 are all regulated through PI3K and Akt activation , making them attractive targets for broad downstream effects . Direct PI3K inhibition has demonstrated impressive tumor inhibition and regression in cell-line and animal models , and multiple agents including DB05210 are currently in clinical trials . Drugs such as perifosine that are specific for Akt are also in development . Combinations of these agents with existing therapies are rational approaches on the path to improving myeloma treatment . Application of HapMap data to the evaluation of 8 candidate genes for pediatric slow transit constipation . BACKGROUND : Slow transit constipation ( P52823 ) affects up to 3 % of all children and is an increasingly recognized cause of chronic constipation in children . We conducted a pilot study to investigate whether genes encoding neurotransmitters ( P20366 , Q9UHF0 , P01282 , NOS1 ) and receptors ( P25103 , P21452 , P29371 , P10721 ) could be responsible for P52823 . METHODS : One hundred seventeen tag single nucleotide polymorphisms ( SNPs ) , distributed among the candidate genes , were selected from HapMap data and genotyped using Sequenom ( San Diego , CA ) technology in 35 affected families . Evaluation of association was performed by transmission disequilibrium test and multilocus analysis . RESULTS : Five SNPs ( rs3771863 , rs4580655 , rs11722288 , rs4563545 , and rs3782221 ) in the P25103 , P29371 , P10721 , and NOS1 genes were found to be potentially associated with P52823 , although the significance of these results does not withstand correction for multiple testing . CONCLUSIONS : Our data indicate that 5 SNPs in the NOS1 , P25103 , P29371 , and P10721 genes could be involved in P52823 , especially rs3771863 in intron 1 of P25103 , which showed the highest association . Enhancement of auranofin-induced apoptosis in MCF-7 human breast cells by selenocystine , a synergistic inhibitor of thioredoxin reductase . P10599 system plays an important role in regulation of intracellular redox balance and various signaling pathways . P30044 ( TrxR ) is overexpressed in many cancer cells and has been identified as a potential target of anticancer drugs . DB00995 ( AF ) is potent TrxR inhibitor with novel in vitro and in vivo anticancer activities . Selenocystine ( SeC ) is a nutritionally available selenoamino acid with selective anticancer effects through induction of apoptosis . In the present study , we demonstrated the synergistic effects and the underlying molecular mechanisms of SeC in combination with AF on MCF-7 human breast cancer cells . The results showed that SeC and AF synergistically inhibited the cancer cell growth through induction of ROS-dependent apoptosis with the involvement of mitochondrial dysfunction . DNA damage-mediated p53 phosphorylation and down-regulation of phosphorylated AKT and P29323 also contributed to cell apoptosis . Moreover , we demonstrated the important role of TrxR activity in the synergistic action of SeC and AF . Taken together , our results suggest the strategy to use SeC and AF in combination could be a highly efficient way to achieve anticancer synergism by targeting TrxR . Leukotriene A4 hydrolase . Inhibition by bestatin and intrinsic aminopeptidase activity establish its functional resemblance to metallohydrolase enzymes . DB03424 , an inhibitor of aminopeptidases , was also a potent inhibitor of leukotriene ( LT ) A4 hydrolase . On isolated enzyme its effects were immediate and reversible with a Ki = 201 +/- 95 mM . With erythrocytes it inhibited LTB4 formation greater than 90 % within 10 min ; with neutrophils it inhibited LTB4 formation by only 10 % during the same period , increasing to 40 % in 2 h . DB03424 inhibited P09960 hydrolase selectively ; neither P09917 nor 15-lipoxygenase activity in neutrophil lysates was affected . Purified P09960 hydrolase exhibited an intrinsic aminopeptidase activity , hydrolyzing L-lysine-p-nitroanilide and L-leucine-beta-naphthylamide with apparent Km = 156 microM and 70 microM and Vmax = 50 and 215 nmol/min/mg , respectively . Both P09960 and bestatin suppressed the intrinsic aminopeptidase activity of P09960 hydrolase with apparent Ki values of 5.3 microM and 172 nM , respectively . Other metallohydrolase inhibitors tested did not reduce P09960 hydrolase/aminopeptidase activity , with one exception ; captopril , an inhibitor of angiotensin-converting enzyme , was as effective as bestatin . The results demonstrate a functional resemblance between P09960 hydrolase and certain metallohydrolases , consistent with a molecular resemblance at their putative Zn2(+)-binding sites . The availability of a reversible , chemically stable inhibitor of P09960 hydrolase may facilitate investigations on the role of LTB4 in inflammation , particularly the process termed transcellular biosynthesis . Nardosinone protects H9c2 cardiac cells from angiotensin II-induced hypertrophy . Pathological cardiac hypertrophy induced by angiotensin II ( AngII ) can subsequently give rise to heart failure , a leading cause of mortality . Nardosinone is a pharmacologically active compound extracted from the roots of Nardostachys chinensis , a well-known traditional Chinese medicine . In order to investigate the effects of nardosinone on AngII-induced cardiac cell hypertrophy and the related mechanisms , the myoblast cell line H9c2 , derived from embryonic rat heart , was treated with nardosinone ( 25 , 50 , 100 , and 200 μmol/L ) or AngII ( 1 μmol/L ) . Then cell surface area and mRNA expression of classical markers of hypertrophy were detected . The related protein levels in PI3K/Akt/ P42345 and MEK/ P29323 signaling pathways were examined by Western blotting . It was found that pretreatment with nardosinone could significantly inhibit the enlargement of cell surface area induced by AngII . The mRNA expression of P01160 , DB04899 and β-MHC was obviously elevated in AngII-treated H9c2 cells , which could be effectively blocked by nardosinone at the concentration of 100 μmol/L . Further study revealed that the protective effects of nardosinone might be mediated by repressing the phosphorylation of related proteins in PI3K/Akt and MEK/ P29323 signaling pathways . It was suggested that the inhibitory effect of nardosinone on Ang II-induced hypertrophy in H9c2 cells might be mediated by targeting PI3K/Akt and MEK/ P29323 signaling pathways . DB01045 -independent interactions between the pregnane X receptor ligand binding domain and peptide fragments of coactivator and corepressor proteins . The pregnane X receptor ( O75469 ) , a member of the nuclear receptor superfamily , regulates the expression of drug-metabolizing enzymes in a ligand-dependent manner . The conventional view of nuclear receptor action is that ligand binding enhances the receptor 's affinity for coactivator proteins , while decreasing its affinity for corepressors . To date , however , no known rigorous biophysical studies have been conducted to investigate the interaction among O75469 , its coregulators , and ligands . In this work , steady-state total internal reflection fluorescence microscopy ( TIRFM ) and total internal reflection with fluorescence recovery after photobleaching were used to measure the thermodynamics and kinetics of the interaction between the O75469 ligand binding domain and a peptide fragment of the steroid receptor coactivator-1 ( Q15788 ) in the presence and absence of the established O75469 agonist , rifampicin . Equilibrium dissociation and dissociation rate constants of ~5 μM and ~2 s(-1) , respectively , were obtained in the presence and absence of rifampicin , indicating that the ligand does not enhance the affinity of the O75469 and Q15788 fragments . Additionally , TIRFM was used to examine the interaction between O75469 and a peptide fragment of the corepressor protein , the silencing mediator for retinoid and thyroid receptors ( Q9Y618 ) . An equilibrium dissociation constant of ~70 μM was obtained for Q9Y618 in the presence and absence of rifampicin . These results strongly suggest that the mechanism of ligand-dependent activation in O75469 differs significantly from that seen in many other nuclear receptors . Glioblastoma : synergy of growth promotion between P13501 and P25103 can be thwarted by blocking P13501 with miraviroc , an FDA approved anti-HIV drug and blocking P25103 with aprepitant , an FDA approved anti-nausea drug . WHAT IS KNOWN AND BACKGROUND : Two receptor signaling pathways that are commonly active in facilitating glioblastoma growth and invasion- that of P51681 and neurokinin ( NK ) -1R- have small molecule inhibitors that are FDA approved and marketed to treat other conditions . The anti-HIV drug , maraviroc , inhibits human P51681 's ligand from binding , and hence blocks P51681 stimulation . The anti-nausea drug aprepitant blocks DB05875 signaling at P25103 . AIMS AND OBJECTIVE : We propose on the basis of molecular insights that a combination of the two drugs is likely to be useful in the treatment of glioblastoma . COMMENT : After stimulation by their respective ligands both P51681 and P25103 , through intermediaries , phosphorylate and thereby activate P27361 /2 , triggering in turn migratory and mitotic events . Neurokinin-1R second messenger signaling also happens to serine phosphorylate P51681 . Phosphorylated P51681 exhibits amplified activity after agonist ligation . Therefore , aprepitant and maraviroc combined treatment is expected to exert synergestic inhibition of growth enhancing signaling in glioblastoma . Inhibiting an amplifier is equivalent to amplifying an inhibitor . Since the two suggested drugs are non-cytotoxic they are envisioned as adjunctive treatments to current standard temozolomide , radiation , and bevacizumab , all to be used after debulking primary resection . WHAT IS NEW AND CONCLUSION : Our analysis makes the case for a well-designed trial of the proposed combination in the treatment of glioblastoma . Contribution of Rho A and Rho kinase to platelet-derived growth factor-BB-induced proliferation of vascular smooth muscle cells . In order to identify small G protein ( s ) which contributes to the proliferation of vascular smooth muscle cells ( VSMCs ) , we examined the effect of an P04035 inhibitor ( cerivastatin ) , a farnesyltransferase inhibitor ( FTI-277 ) , a geranyl geranyl transferase inhibitor ( GGTI-286 ) and a Rho kinase inhibitor ( Y-27632 ) on the proliferation of cultured rat VSMCs stimulated with 20ng/ml platelet-derived growth factor ( PDGF ) -BB . DB00439 and GGTI-286 , but not FTI-277 , suppressed the DB00102 -induced activation of extracellular signal related kinase ( P27361 /2 ) . The inhibitory effect of cerivastatin on the DB00102 -induced activation of P27361 /2 was fully recovered by the addition of geranylgeranyl pyrophosphate ( GGPP ) , but not farnesyl pyrophosphate ( FPP ) . DB00439 and GGTI-286 , but not FTI-277 , suppressed the DB00102 -induced [ 3H ] thymidine incorporation and activation of ornitine decarboxylase ( ODC ) , both of which were fully recovered by the addition of GGPP , but not FPP . These data indicate that the DB00102 -induced activation of P27361 /2 and proliferation of VSMCs depend upon geranylgeranylated small G protein . Immunoblotting analysis revealed the upregulation of Rho A protein in the membrane fractions of VSMCs stimulated by DB00102 . Furthermore , Y-27632 suppressed the DB00102 -induced activation of P27361 /2 and proliferation of VSMCs . On the basis of these data , we conclude that DB00102 stimulates the proliferation of VSMCs via the activation of Rho A . Rho kinase plays an important role in this process as an effector of Rho A . Anti-inflammatory potential of alpha-linolenic acid mediated through selective P36551 inhibition : computational and experimental data . The present work investigates the anti-inflammatory activity of alpha-linolenic acid ( ALA ) and linoleic acid ( LA ) using computational and experimental analysis . The binding affinity of ALA and LA was appraised for cyclooxygenase 1 ( P23219 ) , cyclooxygenase 2 ( P35354 ) , and P09917 ( 5- P28300 ) using AutoDock 4.2 and AutoDock Vina 1.1.2 . Anti-inflammatory activity of ALA ( 2 and 4 ml/kg , i.p. ) ( 55.65 % v/v ) and LA ( 2 and 4 ml/kg , i.p. ) ( 55 % v/v ) was further assayed using the rat paw edema test against a variety of phlogistic agents including carrageenan , arachidonic acid , prostaglandin , and leukotriene , respectively . ALA ( 2 and 4 ml/kg , i.p. ) and LA ( 2 and 4 ml/kg , i.p. ) were further tested for their efficacy against complete Freund 's adjuvant ( O75347 ) -induced ( 0.05 ml ) arthritis in albino rats . Following O75347 -induced arthritis , ALA and LA were tested for their inhibitory proficiency against P23219 , P35354 , and 5- P28300 in vitro . The present study commends that the anti-inflammatory potential of ALA could be attributed to P36551 inhibition , in particular , P35354 . DB00171 -binding cassette transporter A1 R219K polymorphism and ischemic stroke risk in the Chinese population : a meta-analysis . Recently , many studies have been focused on the association between the DB00171 -binding cassette transporter A1 ( O95477 ) gene R219K polymorphism and ischemic stroke ( IS ) . However , the study results have been inconsistent , especially in the Chinese population . Therefore , we performed a meta-analysis to better clarify the association between the O95477 gene and IS . All of the relevant studies used in our meta-analysis were identified using PubMed , OVID , Cochrane Library , Chinese Wan Fang database , Chinese P01282 database , China National Knowledge Infrastructure ( CNKI ) , and China Biological Medicine Database ( CBM ) up to May 2013 . Statistical analysis was conducted with STATA software version 11.0 . Odds ratios with 95 % confidence intervals were applied to evaluate the strength of the association between O95477 gene R219K polymorphism and IS . Heterogeneity was evaluated using the Q-test and I(2) statistic . The funnel plots , Begg 's and Egger 's regression tests were used to assess the publication bias . Our meta-analysis showed the dominant genetic model ( OR=0.92 , 95 % CI : 0.88-0.96 ) , the recessive genetic model ( OR=0.73 , 95 % CI : 0.51-1.05 ) , the homozygote genetic model ( OR=0.64 , 95 % CI : 0.44-0.94 ) , the heterozygote genetic model ( OR=0.81 , 95 % CI : 0.69-0.95 ) , and the allelic genetic model ( OR=0.83 , 95 % CI : 0.69-0.99 ) . For R219K in IS , there were significant associations with these genetic models , but not with the recessive genetic model . Our meta-analysis indicated that the O95477 gene R219K polymorphism might be associated with IS and the K allele might be a protective factor in the Chinese population . DB03843 - or adjuvant-induced peripheral inflammation increases neurokinin-1 receptor gene expression in the mouse . Substance P ( SP ) has been widely studied as a mediator of nociception . The release of SP from primary afferent neurons is increased during nociception , and SP activates neurokinin-1 ( NK-1 ) receptors in the spinal cord and periphery . Nociception-evoked alterations in P25103 gene expression have been studied in rat models of persistent pain but have not been characterized in any murine models of peripheral inflammation . This study assessed behavioral responses and P25103 mRNA gene expression in mice receiving formalin or Freund 's complete adjuvant ( O75347 ) as an inflammatory stimulus . Mechanical withdrawal thresholds were measured before injection of formalin or O75347 and hind paw licking/biting timed during the late-phase of the formalin response . Two and 24 hours after formalin or O75347 injection , mechanical withdrawal thresholds were measured and the mice euthanized . Solution hybridization-nuclease protection assays were used to quantify P25103 mRNA levels . Results demonstrated that inflamed hind paws were edematous , and the withdrawal thresholds of the inflamed hind paws were significantly lower after formalin or O75347 injection . Neurokinin-1 receptor mRNA levels in the ipsilateral dorsal spinal cords of mice were higher at 24 h after formalin injection or 4 days after O75347 injection . These results confirm that mice are hyperalgesic at late time points after formalin or adjuvant injection when P25103 gene expression is elevated in the dorsal spinal cord . This supports the hypothesis that increased P25103 gene expression contributes to the development and maintenance of a hyperalgesic state . Protein kinase DB02527 -dependent regulatory type II beta ( P31323 ) gene variants in antipsychotic-induced weight gain . OBJECTIVE : Antipsychotics are effective in treating schizophrenia symptoms . However , the use of clozapine and olanzapine in particular are associated with significant weight gain . Mouse and human studies suggest that the protein kinase DB02527 -dependent regulatory type II beta ( P31323 ) gene may be involved in energy metabolism , and there is evidence that it is associated with clozapine 's effects on triglyceride levels . We aimed at assessing P31323 's role in antipsychotic-induced weight gain in schizophrenia patients . METHODS : DNA samples from adult schizophrenia or schizoaffective disorder patients of mixed ancestry were genotyped , and weight gain was assessed . We analyzed 16 tag single-nucleotide polymorphisms across the P31323 gene in a Caucasian subset treated either with clozapine or olanzapine ( N = 99 ) . Linear regression based on an additive model was performed with the inclusion of relevant covariates . RESULTS : Normalized per cent weight change was analyzed , revealing that patients with the minor allele at rs9656135 had a mean weight increase of 4.1 % , whereas patients without this allele had an increase of 3.4 % . This association is not significant after correcting for multiple testing . CONCLUSIONS : Because of limited power , P31323 's role in antipsychotic-induced weight gain is unclear , but biological evidence suggests that P31323 may be involved . Further research in larger sample sizes is warranted . Effects of phenacetin and its metabolite p-phenetidine on P23219 and P35354 activities and expression in vitro . The present study was aimed to test the possible cyclooxygenase ( P36551 ) -1/ P35354 selectivity of the old analgesic drug phenacetin and its metabolite p-phenetidine , which exhibits high renal toxicity . DB00316 ( acetaminophen ) , the main metabolite of phenacetin with low renal toxicity , and indomethacin were selected as reference compounds . Collagen-stimulated platelet thromboxane B2 ( TxB2 ) production and phorbol 12-myristate-13-acetate ( PMA ) -induced neutrophil prostaglandin E2 ( DB00917 ) synthesis were used as indicators for P23219 and P35354 activity , respectively . DB03783 was even less potent than paracetamol to reduce the production of both TxB2 and DB00917 , and no clear preference for either of the P36551 -enzymes was seen . P-phenetidine was a more potent inhibitor , already at nanomolar level , of the synthesis of these prostanoids than indomethacin and showed some preference to P35354 inhibition . Somewhat higher , micromolar , concentrations of p-phenetidine also reduced P35354 expression in neutrophils . We suggest that the very potent inhibitory activity of p-phenetidine on DB00917 synthesis combined with the reduction of P35354 expression could explain the renal papillary necrosis in phenacetin kidney .

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DB00898 abuse and HIV infection : role of P14416 . According to a survey from the HIV Cost and Services Utilization Study ( HCSUS ) , approximately 53 % of HIV-infected patients reported drinking alcohol and 8 % were classified as heavy drinkers . The role of alcohol as a risk factor for HIV infection has been widely studied and recent research has found a significant association between heavy alcohol consumption and lower levels of P01730 T cells among HIV-infected alcoholics . Although there is evidence on the role of alcohol as a risk factor for HIV transmission and disease progression , there is a need for population studies to determine the genetic mechanisms that affect alcohol 's role in HIV disease progression . One of the mechanisms of interest is the dopaminergic system . To date , the effects of dopamine on HIV neuroimmune pathogenesis are not well understood ; however , dopaminergic neural degeneration due to HIV is known to occur by viral invasion into the brain via immune cells , and modulation of dopamine in the CNS may be a common mechanism by which different types of substances of abuse impact HIV disease progression . Although previous studies have shown an association of P14416 ( P14416 ) polymorphisms with severity of alcohol dependence , the expression of this allele risk on HIV patients with alcohol dependence has not been systematically explored . In the current study , P14416 Taq1A and C957T SNP genotyping analyses were performed in 165 HIV-infected alcohol abusers and the results were examined with immune status and P01730 counts . 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 . Anti-inflammatory activity of topical THC in DNFB-mediated mouse allergic contact dermatitis independent of P21554 and CB2 receptors . BACKGROUND : ∆(9) - DB00470 ( THC ) , the active constituent of Cannabis sativa , exerts its biological effects in part through the G-protein-coupled P21554 and CB2 receptors , which were initially discovered in brain and spleen tissue , respectively . However , THC also has P21554 /2 receptor-independent effects . Because of its immune-inhibitory potential , THC and related cannabinoids are being considered for the treatment of inflammatory skin diseases . Here we investigated the mechanism of the anti-inflammatory activity of THC and the role of P21554 and CB2 receptors . METHODS : We evaluated the impact of topically applied THC on DNFB-mediated allergic contact dermatitis in wild-type and P21554 /2 receptor-deficient mice . We performed immunohistochemical analyses for infiltrating immune cells and studied the influence of THC on the interaction between T cells , keratinocytes and myeloid immune cells in vitro . RESULTS : Topical THC application effectively decreased contact allergic ear swelling and myeloid immune cell infiltration not only in wild-type but also in P21554 /2 receptor-deficient mice . We found that THC ( 1 ) inhibited the production of IFNγ by T cells , ( 2 ) decreased the production of P13500 and of IFNγ-induced P80075 and CXL10 by epidermal keratinocytes and ( 3 ) thereby limited the recruitment of myeloid immune cells in vitro in a P21554 /2 receptor-independent manner . CONCLUSIONS : Topically applied THC can effectively attenuate contact allergic inflammation by decreasing keratinocyte-derived pro-inflammatory mediators that orchestrate myeloid immune cell infiltration independent of P21554 /2 receptors . This has important implications for the future development of strategies to harness cannabinoids for the treatment of inflammatory skin diseases . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . Pharmacokinetics of riluzole : evidence for glucuronidation as a major metabolic pathway not associated with P22309 genotype . Pharmacokinetic studies of riluzole show a large inter-individual variability of the drug 's clearance and serum concentrations . Optimizing the individual dosage of riluzole may have the potential to improve the effect of riluzole treatment on survival of patients with amyotrophic lateral sclerosis ( P35858 ) . Limited data are available on the in vivo metabolic elimination of riluzole . From in vitro experiments , P05177 seems to be mainly involved in riluzole clearance . However , in vitro studies suggest that formation of riluzole-glucuronide plays a role and may determine the drug 's pharmacokinetic variability in patients to some extent . In the current study the formation of riluzole-glucuronide was examined in amyotrophic lateral sclerosis ( P35858 ) patients . It also aimed at relating glucuronidation of riluzole to differential P22309 *28 genotypes . The formation of riluzole-glucuronide was confirmed in serum from a group of 14 P35858 patients taking riluzole . Riluzole-glucuronide concentrations were positively associated with those of riluzole . In a separate group of 131 P35858 patients taking riluzole , the P22309 *28 genotype was not associated with trough or peak serum concentrations of riluzole . This study provides evidence that the in vivo metabolic elimination of riluzole in P35858 patients involves glucuronidation . The results do not indicate that glucuronidation of riluzole highly contributes to the drug 's inter-individual pharmacokinetic variability . DB00644 antagonist in the management of prostate cancer . DB00044 -releasing hormone ( P01148 ) agonist therapy to induce medical castration has become the most common form of hormonal therapy for advanced and metastatic prostate cancer . When treatment is started , P01148 agonists initially stimulate the release of LH , causing a surge in serum testosterone that can precipitate a " flare " phenomenon or worsening of disease , particularly in patients with bone metastatic disease . DB00644 ( DB00644 ) receptor antagonism represents a newer approach to medical castration . DB00106 is a pure P30968 antagonist that is devoid of any P01148 agonist activity . Results from 1 phase II and 3 phase III clinical trials demonstrate that abarelix produces medical castration more quickly and without causing testosterone surge , as compared with P01148 agonists with or without a nonsteroidal antagonist . The safety profile in terms of adverse events is comparable between the 2 types of treatment , but the lack of testosterone surge with abarelix might confer a safety advantage by abolishing the risk of a disease flare . DB05025 , a coinducer of heat shock proteins for the potential treatment of amyotrophic lateral sclerosis . Recent years have seen an explosion of research into increasingly prevalent neurodegenerative diseases . DB05025 ( BRX-220 ) , being developed by CytRx Corp , is an oral therapeutic candidate for the treatment of amyotrophic lateral sclerosis ( P35858 ) , the most common form of motor neuron disease . P35858 is a fatal , incurable disorder , which can present as sporadic ( 90 to 95 % of cases ) or familial ( 5 to 10 % of cases ) forms . The etiology of sporadic P35858 remains unknown and much of the understanding of P35858 pathogenesis has been derived through study of its familial forms ; in particular , through study of autosomal dominant mutations in the P00441 ( copper/zinc superoxide dismutase ) gene , which cause approximately 20 % of familial P35858 cases . Under conditions of excessive stress , arimoclomol induces amplification of the cytoprotective heat shock response in order to protect motor neurons from death . Comprehensive in vivo and in vitro studies demonstrated its effect in the prevention of neuronal loss and promotion of motor neuron survival , even after symptom onset . Clinical trials have reported good tolerability and safety . This paper discusses the rationale for arimoclomol use in P35858 , the preclinical and clinical evidence collected to date , the likelihood of its promising preclinical results translating to humans , and the relevance of this research for neurodegeneration as a whole . Nitric-oxide synthase knockout modulates Ca²⁺-sensing receptor expression and signaling in mouse mesenteric arteries . Extracellular calcium ( Ca²⁺(e) ) -induced relaxation of isolated , phenylephrine ( PE ) -contracted mesenteric arteries is dependent on an intact perivascular sensory nerve network that expresses the Ca²⁺-sensing receptor ( P41180 ) . Activation of the receptor stimulates an endocannabinoid vasodilator pathway , which is dependent on cytochrome P450 and phospholipase A₂ but largely independent of the endothelium . In the present study , we determined the role of nitric oxide ( NO ) in perivascular nerve P41180 -mediated relaxation of PE-contracted mesenteric resistance arteries isolated from mice . Using automated wire myography , we studied the effects of NO synthase ( NOS ) gene knockout ( NOS(-/-) ) and pharmacologic inhibition of NOS on Ca²⁺(e)-induced relaxation of PE-contracted arteries . P29474 knockout ( P29474 (-/-) ) upregulates but neuronal NOS knockout ( P29475 (-/-) ) downregulates P41180 expression . NOS(-/-) reduced maximum Ca²⁺(e)-induced relaxation with no change in EC₅₀ values , with P29474 (-/-) having the largest effect . The responses of vessels to calindol and Calhex 231 indicate that the P41180 mediates relaxation . L-N⁵-(1-iminoethyl)-ornithine reduced Ca²⁺(e)-induced relaxation of PE-contracted arteries from C57BL/6 control mice by ≈38 % but had a smaller effect in vessels from P29474 (-/-) mice . DB02207 had no significant effect on relaxation of arteries from NOS(-/-) mice , but both N(G)-nitro-L-arginine methylester and N(G)-monomethyl-L-arginine significantly reduced the relaxation maxima in all groups . Interestingly , the P29475 -selective inhibitor S-methyl-L-thiocitrulline significantly increased the EC₅₀ value by ≈60 % in tissues from C57BL/6 mice but reduced the maximum response by ≈80 % in those from P29475 (-/-) mice . Ca²⁺-activated big potassium channels play a major role in the process , as demonstrated by the effect of iberiotoxin . We conclude that P41180 signaling in mesenteric arteries stimulates P29474 and NO production that regulates Ca²⁺(e)-induced relaxation . DB08626 -induced neprilysin inhibition raises amyloid beta levels in rabbit cortex and cerebrospinal fluid . Studies on the pathogenesis of Alzheimer 's disease ( AD ) suggest overproduction of amyloid beta ( Abeta ) may not be the only pathogenic route to AD . Decreased degradation of Abeta is another possible disease mechanism . P08473 is a neutral endopeptidase that has been proposed to be the major enzyme responsible for Abeta degradation . Studies have reported correlations between Abeta deposition and neprilysin activity in the human brain . This study shows that intracerebroventricular infusion of thiorphan , a neprilysin inhibitor , raises cortical and cerebrospinal fluid ( P04141 ) Abeta concentrations in rabbits . Rabbits treated with thiorphan for 5 days had levels of P04141 and cortical Abeta40 that were 147 and 142 % of the control group , respectively . Results for Abeta42 showed a similar trend . The results indicate that age-related decreases of neprilysin could lead to increased brain concentrations of Abeta , plaque formation , and AD . Vitamin D up-regulates the vitamin D receptor by protecting it from proteasomal degradation in human P01730 + T cells . The active form of vitamin D3 , 1,25(OH)2D3 , has significant immunomodulatory properties and is an important determinant in the differentiation of P01730 + effector T cells . The biological actions of 1,25(OH)2D3 are mediated by the vitamin D receptor ( P11473 ) and are believed to correlate with the P11473 protein expression level in a given cell . The aim of this study was to determine if and how 1,25(OH)2D3 by itself regulates P11473 expression in human P01730 + T cells . We found that activated P01730 + T cells have the capacity to convert the inactive DB00146 to the active 1,25(OH)2D3 that subsequently up-regulates P11473 protein expression approximately 2-fold . 1,25(OH)2D3 does not increase P11473 mRNA expression but increases the half-life of the P11473 protein in activated P01730 + T cells . Furthermore , 1,25(OH)2D3 induces a significant intracellular redistribution of the P11473 . We show that 1,25(OH)2D3 stabilizes the P11473 by protecting it from proteasomal degradation . Finally , we demonstrate that proteasome inhibition leads to up-regulation of P11473 protein expression and increases 1,25(OH)2D3-induced gene activation . In conclusion , our study shows that activated P01730 + T cells can produce 1,25(OH)2D3 , and that 1,25(OH)2D3 induces a 2-fold up-regulation of the P11473 protein expression in activated P01730 + T cells by protecting the P11473 against proteasomal degradation . 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 . DB00197 stimulates beta-arrestin-dependent cardiomyocyte contractility via the angiotensin II type 1A receptor . P37231 ( Q07869 gamma ) agonists are commonly used to treat cardiovascular diseases , and are reported to have several effects on cardiovascular function that may be due to Q07869 gamma-independent signaling events . Select angiotensin receptor blockers ( ARBs ) interact with and modulate Q07869 gamma activity , thus we hypothesized that a Q07869 gamma agonist may exert physiologic effects via the angiotensin II type 1(A) receptor ( AT1(A)R ) . In AT1(A)R-overexpressing P29320 293 cells , both angiotensin II ( Ang II ) and the Q07869 gamma agonist troglitazone ( Trog ) enhanced AT1(A)R internalization and recruitment of endogenous beta-arrestin 1/2 ( beta arr1/2 ) to the AT1(A)R . A fluorescence assay to measure diacylglycerol ( DAG ) accumulation showed that although Ang II induced AT1(A)R-G(q) protein-mediated DAG accumulation , Trog had no impact on DAG generation . Trog-mediated recruitment of beta arr1/2 was selective to AT1(A)R as the response was prevented by an ARB- and Trog-mediated beta arr1/2 recruitment to beta1-adrenergic receptor ( beta 1AR ) was not observed . In isolated mouse cardiomyocytes , Trog increased both % and rate of cell shortening to a similar extent as Ang II , effects which were blocked with an ARB . Additionally , these effects were found to be beta arr2-dependent , as cardiomyocytes isolated from beta arr2-KO mice showed blunted contractile responses to Trog . These findings show for the first time that the Q07869 gamma agonist Trog acts at the AT1(A)R to simultaneously block G(q) protein activation and induce the recruitment of beta arr1/2 , which leads to an increase in cardiomyocyte contractility . P42345 regulates the pro-tumorigenic senescence-associated secretory phenotype by promoting P01583 translation . The TOR ( target of rapamycin ) kinase limits longevity by poorly understood mechanisms . DB00877 suppresses the mammalian Q6UUV9 complex , which regulates translation , and extends lifespan in diverse species , including mice . We show that rapamycin selectively blunts the pro-inflammatory phenotype of senescent cells . Cellular senescence suppresses cancer by preventing cell proliferation . However , as senescent cells accumulate with age , the senescence-associated secretory phenotype ( SASP ) can disrupt tissues and contribute to age-related pathologies , including cancer . P42345 inhibition suppressed the secretion of inflammatory cytokines by senescent cells . DB00877 reduced P05231 and other cytokine mRNA levels , but selectively suppressed translation of the membrane-bound cytokine P01583 . Reduced P01583 diminished NF-κB transcriptional activity , which controls much of the SASP ; exogenous P01583 restored P05231 secretion to rapamycin-treated cells . Importantly , rapamycin suppressed the ability of senescent fibroblasts to stimulate prostate tumour growth in mice . Thus , rapamycin might ameliorate age-related pathologies , including late-life cancer , by suppressing senescence-associated inflammation . Control of phenylalanine and tyrosine metabolism by phosphorylation mechanisms . A system for the parallel determination of enzyme phosphorylation and expressed activity in rat liver cells , and its application to studies of phenylalanine hydroxylase and tyrosine aminotransferase , is described . DB00120 hydroxylase is phosphorylated by agents which stimulate cyclic AMP- and Ca2+-dependent protein kinase activity . The phosphorylation site(s) appear to be the same for both kinases . Phosphorylation is accompanied by increased metabolic flux at low , physiologically relevant , substrate concentrations . P01308 and spermine both inhibit the phosphorylation of the enzyme , possibly by increasing dephosphorylation . P17735 is phosphorylated in liver cell incubations but the rate is slow and insensitive to additions to the medium . No parallel changes in flux could be detected . Both enzymes are subject to complex regulatory mechanisms , short- and long-term . Their activities may be coordinated in vivo by control exerted at the level of the plasma membrane where both amino acids share the same transport processes . Determination of the control coefficients for the several components indicates that membrane transport may be a major limitation on flux . Nonlinkage of bipolar illness to tyrosine hydroxylase , tyrosinase , and D2 and D4 dopamine receptor genes on chromosome 11 . OBJECTIVE : Previous linkage and allelic association studies using DNA polymorphisms , cosegregation of cytogenetic abnormalities with psychiatric illness , and assignment of genes involved in neutotransmitter metabolism suggested that chromosome 11 may harbor a gene predisposing to bipolar illness . The authors examined linkage in the families of 14 probands with bipolar illness , with the candidate genes tyrosine hydroxylase ( TH ) , D4 dopamine receptor ( P21917 ) at 11p15 , tyrosinase ( P14679 ) at 11q14-q21 , and D2 dopamine receptor ( P14416 ) at 11q22-q23 , as well as with the c-Harvey-ras oncogene ( P01112 ) and insulin gene ( P01308 ) , both located at 11p15 , a region that previously showed linkage to bipolar illness . METHOD : The genetic data were analyzed with both lod score analysis ( parametric ) and affected-sib-pair analysis ( nonparametric ) ; both narrow and broad definitions of the clinical phenotype were used . Further influences of diagnostic uncertainties were accounted for by using diagnostic probability classes weighing the stability of each phenotype . RESULTS : Two-point linkage results excluded close linkage of bipolar illness to each candidate gene ; negative results were also obtained when the narrow definition of the clinical phenotype was used . Moreover , multipoint linkage analysis of P01112 and P01308 excluded the 11p15 region encompassing both P21917 and TH . In agreement with the negative linkage results , affected-sib-pair analysis did not show preferential sharing of marker alleles at any of the candidate genes . CONCLUSIONS : The negative results obtained under different genetic models exclude a frequent role for P21917 , TH , P14679 , and P14416 in the pathogenesis of bipolar illness . Oral leucine supplementation is sensed by the brain but neither reduces food intake nor induces an anorectic pattern of gene expression in the hypothalamus . DB00149 activates the intracellular mammalian target of the rapamycin ( P42345 ) pathway , and hypothalamic P42345 signaling regulates food intake . Although central infusion of leucine reduces food intake , it is still uncertain whether oral leucine supplementation is able to affect the hypothalamic circuits that control energy balance . We observed increased phosphorylation of p70s6k in the mouse hypothalamus after an acute oral gavage of leucine . We then assessed whether acute oral gavage of leucine induces the activation of neurons in several hypothalamic nuclei and in the brainstem . DB00149 did not induce the expression of Fos in hypothalamic nuclei , but it increased the number of Fos-immunoreactive neurons in the area postrema . In addition , oral gavage of leucine acutely increased the 24 h food intake of mice . Nonetheless , chronic leucine supplementation in the drinking water did not change the food intake and the weight gain of ob/ob mice and of wild-type mice consuming a low- or a high-fat diet . We assessed the hypothalamic gene expression and observed that leucine supplementation increased the expression of enzymes ( P54687 , O15382 and O14874 ) that metabolize branched-chain amino acids . Despite these effects , leucine supplementation did not induce an anorectic pattern of gene expression in the hypothalamus . In conclusion , our data show that the brain is able to sense oral leucine intake . However , the food intake is not modified by chronic oral leucine supplementation . These results question the possible efficacy of leucine supplementation as an appetite suppressant to treat obesity . Polymorphisms associated with egg number at 300 days of age in chickens . We looked for variations that could be associated with chicken egg number at 300 days of age ( EN300 ) in seven genes of the hypothalamic-pituitary-gonadal axis , including gonadotrophin-releasing hormone-I ( DB00644 ) , P30968 ( GnRHR ) , neuropeptide Y ( P01303 ) , dopamine D2 receptor ( P14416 ) , vasoactive intestinal polypeptide ( P01282 ) , P01282 receptor-1 ( VIPR-1 ) , prolactin ( PRL ) , and the QTL region between 87 and 105 cM of the Z chromosome . Ten mutations in the seven genes were chosen to do marker-trait association analyses in a population comprising 1310 chickens , which were obtained from a company located in Guangdong Province of China . The C1704887T of VIPR-1 was found to have a highly significant association with EN300 . The T5841629C of P14416 and the C1715301T of VIPR-1 were significantly associated with EN300 . A highly significant association was also found between the C1704887T-C1715301T haplotypes of VIPR-1 and EN300 . H1H3 had the highest EN300 . Four PCR-RFLP variations in the candidate QTL region were selected to investigate their genetic effects on EN300 . The haplotypes of T32742468C-G32742603A in this region showed a highly significant association with EN300 . Bioinformatics analyses showed that both T32742468C and G32742603A were located in intron 1 of the SH3-domain P62993 -like 2 ( Q99962 ) gene . We conclude that five SNPs , including C1704887T and C1715301T of VIPR-1 , T5841629C of P14416 , and T32742468C and G32742603A of Q99962 , would be useful as markers for breeding to increase chicken EN300 . Lack of association of P35462 and P21554 polymorphisms with premenstrual dysphoric disorders . BACKGROUND : Premenstrual dysphoric disorder ( PMDD ) is a mood disorder characterized with physical and affective symptoms during the luteal phase of susceptible women . OBJECTIVE : The aim of this study was to investigate the association of P35462 ( P35462 ) polymorphism , and Cannabinoid receptor Type 1 ( P21554 ) polymorphism with PMDD . MATERIALS AND METHODS : Fifty one participants with documented PMDD according to the DSM IV criteria and 51 healthy controls were included in this cross sectional study . Symptom severity was measured with daily self-rating , monthly premenstrual assessment forms and psychiatric interviews . The genotyping of P35462 receptor and Cannabinoid type 1 receptors were performed using Taqmanfluorogenic assay method . RESULTS : Distribution of P35462 and P21554 polymorphism was not different between patients and controls . CONCLUSION : These findings do not support a major role of P35462 , and P21554 polymorphisms in contributing to susceptibility to premenstrual dysphoric disorder . A functional variant of the dopamine D3 receptor is associated with risk and age-at-onset of essential tremor . Familial essential tremor ( ET ) , the most common inherited movement disorder , is generally transmitted as an autosomal dominant trait . A genome-wide scan for ET revealed one major locus on chromosome 3q13 . Here , we report that the Ser9Gly variant in the dopamine D(3) receptor gene ( P35462 ) , localized on 3q13.3 , is associated and cosegregates with familial ET in 23 out of 30 French families . Sequencing revealed no other nonsynonymous variants in the P35462 -coding sequence and in the first 871 bp of the 5' flanking region . Moreover , DB00145 -9 homozygous patients presented with more severe and/or earlier onset forms of the disease than heterozygotes . A replication study comparing 276 patients with ET and 184 normal controls confirmed the association of the DB00145 -9 variant with risk and age-at-onset of ET . In human embryonic kidney ( P29320 ) 293-transfected cells , the DB00145 -9 variant did not differ from the DB00133 -9 variant with respect to glycosylation and to anterograde and retrograde trafficking , but dopamine had an affinity that was four to five times higher . With the DB00145 -9 variant , the dopamine-mediated DB02527 response was increased , and the mitogen-associated protein kinase ( MAPK ) signal was prolonged , as compared with the DB00133 -9 variant . The gain-of-function produced by the DB00145 -9 variant may explain why drugs active against tremor in Parkinson 's disease ( PD ) are usually not effective in the treatment of ET and suggests that P35462 partial agonists or antagonists should be considered as novel therapeutic options for patients with ET . Genetics of physical activity and physical inactivity in humans . Emerging evidence suggests that physical activity and sedentary behavior [ reflected in physical inactivity ( PI ) ] , might be two different phenotypes that may have distinct underlying physiological mechanisms . The purpose of this review is to summarize the existing literature on the genetic determinants of PA and PI phenotypes in humans , considering them as distinct behaviors . Completed in January 2011 , this review includes family studies , twin studies , association studies , genome-wide linkage studies and genome-wide association scan ( GWAs ) reporting different physical activity/inactivity-related phenotypes . In regards to PA , familial aggregation studies resulted in heritability estimates ranging from 0 to 60 % , and twin studies yielded heritability estimates ( a(2) ) and shared environment ( c(2) ) scores for PA phenotypes ranging from 0.00 to 0.85 and 0.00 to 0.84 , respectively . Unique environmental ( e(2) ) results are well dispersed from 0.12 to 0.72 . Suggestive linkages were found with markers nearby different activity-related genes : P24530 , P32245 , P25874 , P12104 , P41180 , Q8IVB4 . Significant associations with PA phenotypes were found for Ace , Gln223ARrg , P32245 and P14416 genes . We found one GWAs that reported novel SNPs in the O95340 gene on chromosome 10q23.2 and in two intergenic regions on chromosomes 2q33.1 and 18p11.32 . Heritability estimates for PI ranged from 25 to 60 % and linkage studies recorded higher LOD scores for PI versus PA . The P12821 genotype was strongly associated with PI . There are potentially different genetic influences on PA versus PI phenotypes . Future studies should focus on the different genetic influences on PA and PI to improve our understanding of underlying determinants of these behaviors .

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interacts_with DB00988?

multiple_choice

Aripiprazole : pharmacodynamics of a dopamine partial agonist for the treatment of schizophrenia . Aripiprazole is the first approved atypical antipsychotic with a mechanism of action that exerts a partial agonism with high affinity at DB00988 D2- and Serotonin- P08908 -receptors as well as an antagonism at Serotonin-5- Q13049 -receptors . Aripiprazole provides good clinical effectiveness and a favorable profile of safety and tolerability . The special pharmacodynamics of aripiprazole are described herein . Methodological challenges in monitoring new treatments for rare diseases : lessons from the cryopyrin-associated periodic syndrome registry . BACKGROUND : The Q96P20 -Associated Periodic Syndromes ( CAPS ) are a group of rare hereditary autoinflammatory diseases and encompass Familial Cold Autoinflammatory Syndrome ( FCAS ) , Muckle-Wells Syndrome ( MWS ) , and Neonatal Onset Multisystem Inflammatory Disease ( NOMID ) . DB06168 is a monoclonal antibody directed against P01584 and approved for CAPS patients but requires post-approval monitoring due to low and short exposures during the licensing process . Creative approaches to observational methodology are needed , harnessing novel registry strategies to ensure Health Care Provider reporting and patient monitoring . METHODS : A web-based registry was set up to collect information on long-term safety and effectiveness of canakinumab for CAPS . RESULTS : Starting in November 2009 , this registry enrolled 241 patients in 43 centers and 13 countries by December 31 , 2012 . One-third of the enrolled population was aged < 18 ; the overall population is evenly divided by gender . Enrolment is ongoing for children . CONCLUSIONS : Innovative therapies in orphan diseases require post-approval structures to enable in depth understanding of safety and natural history of disease . The rarity and distribution of such diseases and unpredictability of treatment require innovative methods for enrolment and follow-up . Broad international practice-based recruitment and web-based data collection are practical . Large-cell neuroendocrine carcinoma of the ampulla of Vater . Large-cell neuroendocrine carcinoma is a high-grade neuroendocrine carcinoma , originally described in the lung . The tumor rarely occurs in extrapulmonary sites like the gastrointestinal tract , and only few examples have been described in the ampulla of Vater . A new case of large-cell neuroendocrine carcinoma of the ampulla of Vater in a 60-year-old man is reported . After pancreatoduodenectomy , macroscopic examination revealed ulcerated tumor in the region of the ampulla of Vater . Microscopically , the tumor exhibited organoid , predominantly nested growth pattern , consisting of large , polygonal cells with pleomorphic nuclei . Average number of mitoses was 36 per 10 high-power fields . Small and large areas of necrosis were identified . Immunohistochemically , the tumor cells were positive for synaptophysin , chromogranin A , P09936 , neuron-specific enolase , pancytokeratin , CK8 and somatostatin and negative for CK7 , CK20 , S-100 , Q15669 -1 , HMB-45 , CD117 , P12830 and regulatory peptides . Ki-67 proliferative index was 41 % . Histone deacetylase ( HDAC ) analysis showed almost identical results for Q13547 , Q92769 and O15379 -- 60 , 60.3 and 61 % , respectively . Two months after surgery , liver metastases occurred , confirming highly aggressive behavior of large-cell neuroendocrine carcinoma . 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 . The antiproliferative effect of lidocaine on human tongue cancer cells with inhibition of the activity of epidermal growth factor receptor . Local anesthetics suppress proliferation in several cancer cells . The mechanism of the suppression , however , is unknown . Our previous study shows that lidocaine , at the level of tissue concentration under topical or local administration , has a direct inhibitory effect on the activity of epidermal growth factor receptor ( P00533 ) , which is a potential target for antiproliferation in cancer cells . Therefore , we hypothesized that lidocaine would suppress the proliferation of cancer cells through the inhibition of P00533 activity . We investigated the effects of lidocaine ( 40-4000 microM ) on proliferation of a human tongue cancer cell line , CAL27 , which has a high level of P00533 expression , and also examined the effect of lidocaine on epidermal growth factor ( P01133 ) -stimulated autophosphorylation of P00533 in CAL27 cells . A clinical concentration of lidocaine ( 400 microM ) suppressed both serum-induced and P01133 -induced proliferation of CAL27 cells and inhibited P01133 -stimulated tyrosine kinase activity of P00533 without cytotoxicity . A larger concentration of lidocaine ( 4000 microM ) showed cytotoxicity with an antiproliferative effect . We suggest that the inhibition of P01133 -stimulated P00533 activity is one of the mechanisms of the antiproliferative effect of lidocaine on CAL27 cells . DB00281 administered topically within the oral cavity for cancer pain relief may suppress the proliferation of human tongue cancer cells . Targeting the epigenome in the treatment of asthma and chronic obstructive pulmonary disease . Epigenetic modification of gene expression by methylation of DNA and various post-translational modifications of histones may affect the expression of multiple inflammatory genes . Acetylation of histones by histone acetyltransferases activates inflammatory genes , whereas histone deacetylation results in inflammatory gene repression . Corticosteroids exert their antiinflammatory effects partly by inducing acetylation of antiinflammatory genes , but mainly by recruiting histone deacetylase-2 ( Q92769 ) to activated inflammatory genes . Q92769 deacetylates acetylated glucocorticoid receptors so that they can suppress activated inflammatory genes in asthma . In chronic obstructive pulmonary disease ( P48444 ) , there is resistance to the antiinflammatory actions of corticosteroids , which is explained by reduced activity and expression of Q92769 . This can be reversed by a plasmid vector , which restores Q92769 levels , but may also be achieved by low concentrations of theophylline . Oxidative stress causes corticosteroid resistance by reducing Q92769 activity and expression by activation of phosphoinositide-3-kinase-delta , resulting in Q92769 phosphorylation via a cascade of kinases . DB00277 reverses corticosteroid resistance by directly inhibiting oxidant-activated O00329 and is mimicked by O00329 knockout or by selective inhibitors . Other treatments may also interact in this pathway , making it possible to reverse corticosteroid resistance in patients with P48444 , as well as in smokers with asthma and some patients with severe asthma in whom similar mechanisms operate . Other histone modifications , including methylation , tyrosine nitration , and ubiquitination may also affect histone function and inflammatory gene expression , and better understanding of these epigenetic pathways could led to novel antiinflammatory therapies , particularly in corticosteroid-resistant inflammation . PEGylation of growth hormone-releasing hormone ( P01286 ) analogues . Synthetically produced GRF1-29 ( DB00010 ) has an amino acid composition identical to the N-terminal 29 amino acids sequence of the natural hypothalamic GHRH1-44 ( Figure 1 ) . It maintains bioactivity in vitro and is almost equally effective in eliciting secretion of endogenous growth hormone in vivo . The main drawbacks associated with the pharmaceutical use of hGRF1-29 relate to its short half-life in plasma , about 10-20 min in humans , which is caused mostly by renal ultrafiltration and enzymatic degradation at the N terminus . PEGylation has been considered as one valid approach to obtain more stable forms of the peptide , with a longer in vivo half-life and ultimately with increased pharmacodynamic response along the somatotropic axis ( endogenous GH , DB01277 levels ) . Different PEGylated P01286 conjugates were obtained and their bioactivity was tested in vitro and in vivo by monitoring endogenous growth hormone ( GH ) serum levels after intravenous ( i.v. ) injection in rats , and intravenous and subcutaneous ( s.c. ) injection in pigs . It was found that P01286 -PEG conjugates are able to bind and activate the human Q02643 , although with different potency . The effect of PEG molecular weight , number of PEG chains bound and position of PEGylation site on P01286 activity were investigated . Mono-PEGylated isomers with a PEG5000 polymer chain linked to Lys 12 or Lys 21 residues , showed high biological activity in vitro , which is similar to that of hGRF1-29 , and a higher pharmacodynamic response as compared to unmodified P01286 molecule . Role of the P08908 receptor in development of the neonatal rat brain : preliminary behavioral studies . Serotonin exerts an influence on the prenatal development of rat brain . However , later developmental times may be more applicable to the understanding of the role of serotonin in human developmental disorders . Therefore , the current study was undertaken to gain preliminary information on the postnatal effects of serotonin on rat brain development . As the P08908 receptor has been shown to be involved in much of the developmental functions of serotonin , an agonist for this receptor , 8-hydroxy-DPAT ( 8-OH-DPAT ) , was used . Neonatal rat pups at three ages ( postnatal days , PNDs ) 3-10 , 10-17 or 17-24 ) were injected daily with 1 mg/kg 8-OH-DPAT and evaluated for behavioral consequences . The youngest group showed accelerated incisor eruption and eye-opening , a possible consequence of P08908 receptor interactions with epidermal growth factor ( P01133 ) . Behaviorally , the animals were more anxious . Animals treated from P01160 10-17 , showed no change in craniofacial development but showed greater behavioral maturity in measures of spontaneous alternation and activity in the open field . The oldest animals ( P01160 17-24 ) showed no behavioral alterations , suggesting that this time length is beyond the critical period for serotonin 's influence in brain development . [ DB00988 -beta-hydroxylaseaktivität im Plasma von Dialysepatienten ( author 's transl ) ] . Plasma dopamin-b-hydroxylase ( P09172 ) was studied in 70 healthy control persons and in 37 hemodialysed patients . Basal P09172 in controls corresponded to 50.0 +/- 29.3 IU . There was was no significant difference between males ( 53.9 +/1 33.8 IU ) and females ( 47.4 +/- 25 IU ) ; no correlation could be found between age and plasma P09172 . In hemodialysed patients basal P09172 levels were significantly ( p less than 0.01 ) decreased ( 32.5 % /- 17.6 IU ) , suggesting lowered sympathetic activity and/or abnormalities in release , distribution space , or metabolism of P09172 . During hemodialysis plasma P09172 activity rose during ultrafiltration . This finding indicates a directionally appropriate sympathetic reflex response to volume depletion in dialysed patients . Use of P01148 antagonists in reproductive medicine . Gonadotrophin-releasing hormone ( DB00644 ) plays a key role in the secretion of gonadotrophins , follicle-stimulating hormone ( DB00094 ) and luteinizing hormone ( LH ) , which regulate steroidogenesis and folliculogenesis . Two DB00644 antagonists , DB00050 and DB06785 , deprived of histaminergic side-effects , have been introduced into ovarian stimulation protocols to prevent premature LH surges and proved their safety in clinical trials . At present , most of the published studies have not found significant differences in follicular recruitment , oocyte quality , and so on , except for a decrease in pregnancy and implantation rates in in vitro fertilization and embryo transfer ( IVF-ET ) cycles when the DB00644 antagonist rather than the agonist was used . This decrease in pregnancy rates was in relation with a necessary learning curve of the physicians . Another possibility is the impact of the DB00644 antagonist on endometrium through its P30968 ; this effect was cancelled after cryopreserved embryo transfers because the pregnancy rates were similar between DB00644 antagonist and agonist in this case . DB00644 antagonists were also interesting in poor responders and polycystic ovarian syndrome , where the agonists have not permitted to obtain the better results in IVF-ET cycles . Similarly , the DB00644 antagonists could prevent the LH surge in the intrauterine insemination cycles . Effect of hydrocortisone on the pituitary response to growth hormone releasing hormone . RATIONALE : In depression , the growth hormone ( GH ) response to clonidine and L-tryptophan ( L-TRP ) is reduced , suggesting reduced alpha2-adrenergic and serotonin (5-HT)1A receptor function . Pretreatment with hydrocortisone ( 100 mg , orally 11 h before ) also blunts the GH response to L-TRP . This effect may be mediated at the hypothalamic level via reduced P08908 receptor function or at the pituitary level , either by a direct effect on somatotrope cells or via enhanced insulin-like growth factor-1 ( DB01277 ) or somatostatin ( SS ) release . OBJECTIVES : To examine the effects of acute and chronic exposure to hydrocortisone on baseline and stimulated GH release from the pituitary . METHODS : Twelve healthy male volunteers received pretreatment with acute hydrocortisone ( 100 mg , 11 h before ) , chronic hydrocortisone ( 20 mg twice a day for 1 week ) and placebo in a double blind , balanced order , crossover design . Serial measurements of plasma GH , DB01277 and thyroid stimulating hormone ( DB00024 ) levels were made at baseline and following intravenous administration of 1 mcg/kg P01286 . RESULTS : The GH response to growth hormone releasing hormone ( P01286 ) was significantly blunted by pretreatment with both acute and chronic hydrocortisone . Baseline DB01277 levels were significantly lower at baseline after chronic hydrocortisone compared with placebo . Baseline DB00024 levels were significantly lower after acute hydrocortisone compared with placebo , suggesting an increase in somatostatin levels . CONCLUSIONS : These data suggest that hydrocortisone acts at the pituitary level to reduce GH release . The DB00024 and DB01277 data support the hypothesis that hydrocortisone reduces GH release by enhancing somatostatin and DB01277 release . Stimulatory effects of 5HT1A receptor agonists on luteinizing hormone-releasing hormone release from cultured fetal rat hypothalamic cells : interactions with progesterone . Previous works have suggested an interactive stimulatory effect of progesterone ( P ) and serotonin ( 5-HT ) on luteinizing hormone release . The purpose of the present study was to determine whether 5-HT via P08908 receptors interacts with P in the process of luteinizing hormone-releasing hormone ( P01148 ) release . Using fetal hypothalamic neurons in primary cell cultures the first goal of this study was to determine the effects of P08908 receptor agonists on P01148 secretion . 8-Hydroxy-2 ( di-n-propylamino ) tetralin ( 8-OH-DPAT ) or ipsapirone ( 10(-5) M ) significantly stimulated P01148 release . Pharmacological studies have allowed to rule out the possible involvement of alpha 2- or beta-adrenoreceptors , or 5-HT uptake sites , in the stimulatory effect of 8-OH-DPAT on P01148 release , thus demonstrating the specific involvement of P08908 receptors in the stimulation of P01148 release . The second goal was to test the ability of P to stimulate P01148 release from fetal hypothalamic neurons . P ( 10(-6) M ) applied for 30 or 120 min significantly stimulated P01148 secretion . The maintenance of the stimulation of P01148 release by P after a cycloheximide treatment or by an impermeable analog of P , P-3-BSA , has suggested a nongenomic effect of P on P01148 release . The effects of a pretreatment of cells by P on 8-OH-DPAT-induced P01148 release were tested . While 10(-7) M P alone did not stimulate P01148 release , this concentration of steroid potentiated the P01148 response to 10(-5) M 8-OH-DPAT . These findings led to the conclusion that P acting at the level of the plasma membrane potentiates the stimulatory effect of P08908 receptor agonists on P01148 release . Anti-angiogenic agent ramucirumab : meaningful or marginal ? DB05578 ( IMC-1121B ) targets P35968 . DB05578 is being investigated in many malignancies including gastric cancer . The Phase III trial in patients with advanced breast cancer failed to improve the primary end point The REGARD trial , a Phase III study , in patients with advanced gastric cancer in the second line setting , had a marginal improvement in overall survival but did not achieve the expected hazard ratio target ( of 0.69 ) and the median duration of therapy with ramucirumab was meager 8 weeks ( only 2 weeks longer than the placebo 's ) . Other notable agents in the second line setting are docetaxel and irinotecan . Preliminary results of the RAINBOW trial suggest that ramucirumab may be providing more than marginal advantage . In this review , we briefly summarize the process of angiogenesis and address the emerging cost-benefit issues that surround all newly developed agents including ramucirumab . The N676D and G697R mutations in the kinase domain of P36888 confer resistance to the inhibitor DB05213 . P55008 arrest and down-regulation of cyclin E/cyclin-dependent kinase 2 by the protein kinase inhibitor staurosporine are dependent on the retinoblastoma protein in the bladder carcinoma cell line 5637 . The protein kinase inhibitor staurosporine has been shown to induce P55008 phase arrest in normal cells but not in most transformed cells . DB02010 did not induce P55008 phase arrest in the bladder carcinoma cell line 5637 that lacks a functional retinoblastoma protein ( pRB- ) . However , when infected with a pRB-expressing retrovirus [ Goodrich , D. W. , Chen , Y. , Scully , P. & Lee , W.-H. ( 1992 ) Cancer Res. 52 , 1968-1973 ] , these cells , now pRB+ , were arrested by staurosporine in P55008 phase . This arrest was accompanied by the accumulation of hypophosphorylated pRB . In both the pRB+ and pRB- cells , cyclin D1-associated kinase activities were reduced on staurosporine treatment . In contrast , cyclin-dependent kinase ( CDK ) 2 and cyclin E/ P24941 activities were inhibited only in pRB+ cells . DB02010 treatment did not cause reductions in the protein levels of P11802 , cyclin D1 , P24941 , or cyclin E . The CDK inhibitor proteins P38936 (Waf1/Cip1) and p27 ( Kip1 ) levels increased in staurosporine-treated cells . Immunoprecipitation of P24941 , cyclin E , and p2l from staurosporine-treated pRB+ cells revealed a 2.5- to 3-fold higher ratio of p2l bound to P24941 compared with staurosporine-treated pRB- cells . In pRB+ cells , p2l was preferentially associated with Thrl6O phosphorylated active P24941 . In pRB- cells , however , p2l was bound preferentially to the unphosphorylated , inactive form of P24941 even though the phosphorylated form was abundant . This is the first evidence suggesting that P55008 arrest by 4 nM staurosporine is dependent on a functional pRB protein . Cell cycle arrest at the pRB- dependent checkpoint may prevent activation of cyclin E/ P24941 by stabilizing its interaction with inhibitor proteins p2l and p27 . Genetic basis of delay discounting in frequent gamblers : examination of a priori candidates and exploration of a panel of dopamine-related loci . INTRODUCTION : Delay discounting is a behavioral economic index of impulsivity that reflects preferences for small immediate rewards relative to larger delayed rewards . It has been consistently linked to pathological gambling and other forms of addictive behavior , and has been proposed to be a behavioral characteristic that may link genetic variation and risk of developing addictive disorders ( i.e. , an endophenotype ) . Studies to date have revealed significant associations with polymorphisms associated with dopamine neurotransmission . The current study examined associations between delay discounting and both previously linked variants and a novel panel of dopamine-related variants in a sample of frequent gamblers . METHODS : Participants were 175 weekly gamblers of European ancestry who completed the Monetary Choice Questionnaire to assess delay discounting preferences and provided a DNA via saliva . RESULTS : In a priori tests , two loci previously associated with delayed reward discounting ( rs1800497 and rs4680 ) were not replicated , however , the long form of P21917 VNTR was significantly associated with lower discounting of delayed rewards . Exploratory analysis of the dopamine-related panel revealed 11 additional significant associations in genes associated with dopamine synthesis , breakdown , reuptake , and receptor function ( P35462 , Q01959 , DDC , P09172 , and Q05940 ) . An aggregate genetic risk score from the nominally significant loci accounted for 17 % of the variance in discounting . Mediational analyses largely supported the presence of indirect effects between the associated loci , delay discounting , and pathological gambling severity . CONCLUSIONS : These findings do not replicate previously reported associations but identify several novel candidates and provide preliminary support for a systems biology approach to understand the genetic basis of delay discounting . Xaliproden ( SR57746A ) induces P08908 receptor-mediated Q96HU1 kinase activation in PC12 cells . Neurotrophic growth factors are involved in cell survival . However , natural growth factors have a very limited therapeutic use because of their short half-life . In the present study , we investigated the mechanism of action of a non-peptidic neurotrophic drug , Xaliproden , a potential molecule for the treatment of motoneuron diseases , since the transduction pathways of this synthetic P08908 agonist are very poorly understood . Xaliproden does not activate the Trk receptor but causes a rapid increase in the activities of the P27361 and P28482 isoforms of Q96HU1 kinase , which then rapidly decrease to the basal level . We demonstrate that isoforms of the P29353 adapter protein are phosphorylated independently of each other and are probably not the source of the Xaliproden-induced Q96HU1 kinases activation . The inhibitor of Ras farnesylation , FPT-1 , and the protein kinase C inhibitors , GF 109203X and chelerythrine , inhibited the Xaliproden-induced Q96HU1 kinase activation , suggesting p21Ras and PKC involvement . Moreover , the observations that the P08908 antagonist , pindobind , and pertussis toxin abolished the Xaliproden-induced P29323 stimulation suggested that Xaliproden activates the Q96HU1 kinase pathways by stimulating the G protein-coupled receptor , P08908 . These results demonstrate clearly that the non-peptidic compound , Xaliproden , exerts its neurotrophic effects through a mechanism of action differing from that of neurotrophins . These findings suggest that this compound does not involve MAPK activation by TrkA receptor stimulation but acts by Q96HU1 kinase pathway by a pertussis toxin-sensitive mechanism involving P08908 receptors , P38936 Ras and MEK-1 and by PKC and Akt pathways . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Raddeanin A , a triterpenoid saponin isolated from Anemone raddeana , suppresses the angiogenesis and growth of human colorectal tumor by inhibiting P35968 signaling . Raddeanin A ( RA ) is an active triterpenoid saponin from a traditional Chinese medicinal herb , Anemone raddeana Regel . It was previously reported that RA possessed attractive antitumor activity through inhibiting proliferation and inducing apoptosis of multiple cancer cells . However , whether RA can inhibit angiogenesis , an essential step in cancer development , remains unknown . In this study , we found that RA could significantly inhibit human umbilical vein endothelial cell ( HUVEC ) proliferation , motility , migration , and tube formation . RA also dramatically reduced angiogenesis in chick embryo chorioallantoic membrane ( P62158 ) , restrained the trunk angiogenesis in zebrafish , and suppressed angiogenesis and growth of human HCT-15 colorectal cancer xenograft in mice . Western blot assay showed that RA suppressed P15692 -induced phosphorylation of P35968 and its downstream protein kinases including PLCγ1 , O60674 , Q05397 , Src , and Akt . Molecular docking simulation indicated that RA formed hydrogen bonds and hydrophobic interactions within the DB00171 binding pocket of P35968 kinase domain . Our study firstly provides the evidence that RA has high antiangiogenic potency and explores its molecular basis , demonstrating that RA is a potential agent or lead candidate for antiangiogenic cancer therapy . Lack of biological relevance of platelet cyclooxygenase-2 dependent thromboxane A2 production . INTRODUCTION : There is emerging evidence of a considerable variability of the impact of aspirin on clinical outcome and laboratory findings . Persistent TxA2 production seems to be the most likely reason . Aim of this study was to determine whether the mechanism responsible for TxA2 persistent production is , at least partially , dependent upon aspirin-insensitive platelet P35354 enzymatic pathway . METHODS AND RESULTS : In 100 consecutive patients , under chronic aspirin anti-platelet treatment ( 100-160 mg/day ) selected on the basis of detectable plasma salicylate levels , serum and DB04557 ( AA ) -induced platelet TxA2 production , immunoblot analysis of platelet P23219 / P35354 expression and P35354 activity were studied . Immunoblot revealed P35354 expression in 46 % patients , in an amount that was markedly lower than P23219 . In 10 P35354 positive patients with TxA2 levels over the median , AA-induced TxA2 production performed in vitro in the presence of the P35354 inhibitor CAY10404 and aspirin demonstrated that P35354 dependent TxA2 production is less than 2 % . CONCLUSION : Our data demonstrate that the inter-individual variability of platelet sensitivity to aspirin is due to a reduced efficacy of aspirin on platelet P23219 despite ascertained patient compliance . We suggest that serum TxA2 assay might be performed in future clinical studies to improve our knowledge on the residual TxA2 production in aspirin-treated patients . Signaling pathways mediating induction of the early response genes prostaglandin G/H synthase-2 and egr-1 by serotonin via 5- Q13049 receptors . Signaling pathways responsible for serotonin ( 5-HT ) -mediated induction of early response genes prostaglandin G/H synthase-2 ( P35354 , cyclooxygenase-2 ) and egr-1 were investigated in rat mesangial cells . Gene induction by 5-HT was dependent on 5- Q13049 receptors that were pertussis toxin insensitive indicating coupling to a G-protein of the Gq family . Binding of 5-HT to this receptor activates phosphatidylinositol-specific phospholipase C ( P98160 ) and release of Ca2+ from internal stores , but this activation was not related to P35354 mRNA expression . Similarly , P19957 kinase was not involved in 5-HT signaling . Instead , inhibition of phosphatidylcholine-specific P98160 interfered with P35354 and egr-1 mRNA induction , suggesting this enzyme as a link between 5- Q13049 receptors and protein kinase C , an essential part of 5-HT-mediated signaling . The Q96HU1 kinase pathway was identified as common signaling pathway of 5-HT or phorbol ester-induced gene expression . Increase of intracellular DB02527 by forskolin or dibutyryl DB02527 did not induce P35354 or egr-1 mRNA expression by itself , but strongly inhibited 5-HT-mediated mRNA induction . P35354 mRNA and protein induction by 5-HT was also abolished by chelation of Ca2+ ions by EGTA , suggesting involvement of Ca2+-dependent enzymes . In contrast , egr-1 mRNA expression was superinduced in the presence of EGTA . Induction of Egr-1 protein was not changed by EGTA hinting to Ca2+-sensitive posttranscriptional steps . Activation of the Gq-coupled 5- Q13049 receptor thus leads to the expression of the early response genes P35354 and egr-1 , using common as well as differing signaling elements that allow differential regulation of the expression of these genes that are functionally related to renal hemodynamics and proliferation of mesangial cells , respectively . Chemical coding of the human gastrointestinal nervous system : cholinergic , VIPergic , and catecholaminergic phenotypes . The aim of this investigation was to identify the proportional neurochemical codes of enteric neurons and to determine the specific terminal fields of chemically defined nerve fibers in all parts of the human gastrointestinal ( GI ) tract . For this purpose , antibodies against the vesicular monoamine transporters ( P54219 /2 ) , the vesicular acetylcholine transporter ( Q16572 ) , tyrosine hydroxylase ( TH ) , dopamine beta-hydroxylase ( P09172 ) , serotonin ( 5-HT ) , vasoactive intestinal peptide ( P01282 ) , and protein gene product 9.5 ( P09936 ) were used . For in situ hybridization (35)S-labeled P54219 , Q05940 , and Q16572 riboprobes were used . In all regions of the human GI tract , 50-70 % of the neurons were cholinergic , as judged by staining for Q16572 . The human gut unlike the rodent gut exhibits a cholinergic innervation , which is characterized by an extensive overlap with VIPergic innervation . Neurons containing Q05940 constituted 14-20 % of all intrinsic neurons in the upper GI tract , and there was an equal number of TH-positive neurons . In contrast , P09172 was absent from intrinsic neurons . Cholinergic and monoaminergic phenotypes proved to be completely distinct phenotypes . In conclusion , the chemical coding of human enteric neurons reveals some similarities with that of other mammalian species , but also significant differences . P01282 is a cholinergic cotransmitter in the intrinsic innervation of the human gut . The substantial overlap between Q05940 and TH in enteric neurons indicates that the intrinsic catecholaminergic innervation is a stable component of the human GI tract throughout life . The absence of P09172 from intrinsic catecholaminergic neurons indicates that these neurons have a dopaminergic phenotype . Preferential expression of the vasoactive intestinal peptide ( P01282 ) receptor P32241 in human cord blood-derived P28906 + P28907 - cells : possible role of P01282 as a growth-promoting factor for hematopoietic stem/progenitor cells . Primitive hematopoietic progenitor cells such as severe combined immunodeficiency- repopulating cells and long-term culture-initiating cells are enriched in P28906 + P28907 - cells derived from various stem cell sources . In this study , to elucidate the features of such primitive cells at the molecular level , we tried to isolate genes that were preferentially expressed in umbilical cord blood ( CB ) -derived P28906 + P28907 - cells by subtractive hybridization . The gene for P32241 receptor , a receptor for the neuropeptide vasoactive intestinal peptide ( P01282 ) , was thereby isolated and it was shown that this gene was expressed in both P28906 + P28907 - and P28906 + P28907 + CB cells and that the expression levels were higher in P28906 + P28907 - CB cells . Next , we assessed the effects of P01282 on the proliferation of P28906 + CB cells using in vitro culture systems . In serum-free single-cell suspension culture , P01282 enhanced clonal growth of P28906 + CB cells in synergy with P36888 ligand ( FL ) , stem cell factor ( P21583 ) , and thrombopoietin ( P07202 ) . In serum-free clonogenic assays , P01282 promoted myeloid ( colony-forming unit-granulocyte/macrophage ( CFU-GM ) ) and mixed ( CFU-Mix ) colony formations . Furthermore , in Dexter-type long-term cultures , P01282 increased colony-forming cells at week 5 of culture . These results suggest that P01282 functions as a growth-promoting factor of CB-derived hematopoetic progenitor cells .

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interacts_with DB00912?

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Genetics of type 2 diabetes mellitus and other specific types of diabetes ; its role in treatment modalities . Type 2 diabetes mellitus ( T2DM ) is among the most challenging health issues of the 21st century and is associated with an alarming rise in the incidence . The pathophysiological processes that lead to development of T2DM are still unclear , however impairment in insulin secretion and/or action is clearly indicated . Type 2 diabetes is a polygenic disorder with multiple genes located on different chromosomes contributing to its susceptibility . Analysis of the genetic factors is further complicated by the fact that numerous environmental factors interact with genes to produce the disorder . Only a minority of cases of type 2 diabetes are caused by single gene defects and one example is maturity onset diabetes of the young ( MODY ) . Previous studies indicated that variants in genes encoding the pancreatic β-cell K+ DB00171 channel subunits Kir6.2 ( Q14654 ) and Q09428 ( Q09428 ) are associated with neonatal diabetes . Six different types of maturity onset diabetes of young ( MODY ) have been identified based on characteristic gene defect . The common Pro12Ala polymorphism in peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) gene was confirmed in several studies to be associated with type 2 diabetes as well . More recently , studies reported variants within a novel gene , Q9NQB0 , as a putative susceptibility gene for type 2 diabetes across many ethnic backgrounds around the world . MODY patients respond better to sulphonylureas and metformin , while neonatal diabetes patients with genetic mutations can be changed from insulin to oral drugs . We hereby provide a comprehensive review on the role of genetics in type 2 diabetes mellitus . Endothelial Q09428 -8 acts in an P29323 -independent pathway during atrioventricular cushion development . Q09428 -8 , a conserved leucine-rich repeats protein , was first identified as a positive regulator of Ras pathway in Caenorhabditis elegans . Biochemical studies indicated that Q09428 -8 interacts with Ras and Raf , leading to the elevated P29323 activity . However , the physiological role of Q09428 -8 during mammalian development remains unclear . Here we found that germline deletion of Q09428 -8 in mice resulted in early embryonic lethality . Inactivated Q09428 -8 specifically in mouse endothelial cells ( ECs ) revealed that Q09428 -8 is essential for embryonic heart development . Q09428 -8 deficiency in ECs resulted in late embryonic lethality , and the mutant mice displayed multiple cardiac defects . The reduced endothelial-mesenchymal transformation ( EMT ) and the reduced mesenchyme proliferation phase were observed in the atrioventricular canal ( AVC ) within the mutant hearts , leading to the formation of hypoplastic endocardial cushions . However , P29323 activation did not appear to be affected in mutant ECs , suggesting that Q09428 -8 may act in an P29323 -independent pathway to regulate AVC development . Metronomic chemotherapy dosing-schedules with estramustine and temozolomide act synergistically with anti- P35968 antibody to cause inhibition of human umbilical venous endothelial cell growth . The effects of ' metronomic ' or extended chemotherapy dosing schedules ( ECS ) are mediated through poorly understood anti-angiogenic mechanisms . ECS combined with biological anti-angiogenic agents have produced promising pre-clinical results . MATERIALS AND METHODS : We have expanded the list of agents with an in vitro ECS profile to include the methylating agent temozolomide ( DB00853 ) and the anti-mitotic agent estramustine ( Estracyt ) . These agents were also combined with a specific anti-angiogenic inhibitor DB06101 and a non-specific agent with anti-angiogenic properties , Compound 5h . The in vitro HUVEC ECS model system was optimised and cell proliferation assays undertaken . RESULTS : As a single agent , estramustine inhibited endothelial cell proliferation with an IC50 of 4.5 microM and was active at 10-33 % of the maximum tolerated dose ( MTD ) from clinical schedules , whilst temozolomide had IC50 of 6.6 microM and was active at 1-6 % of MTD . In combination , significant synergy was seen with DB06101 in combination with either drug , whilst modest additive effects were observed with Compound 5h . None of the combinations resulted in significant cytotoxicity or apoptosis . DISCUSSION : The results show that ECS of temozolomide and estramustine can be significantly enhanced when combined with specific anti-angiogenic inhibitors in an in vitro HUVEC system . DB05258 receptor 2 expression by peripheral blood monocytes in patients with a high viral load of hepatitis C virus genotype 1 showing substitution of amino Acid 70 in the core region . BACKGROUND/AIM : When patients with chronic hepatitis C ( CHC ) are treated with interferon ( IFN ) -based therapy , achieving serum HCV-RNA negativity by week 12 ( early viral response , EVR ) is an important predictor of a sustained virologic response . The aim of this study was to clarify whether changes in IFN-alpha receptor 2 ( P17181 -2 ) expression by peripheral blood monocytes ( Mo ) and the EVR rate differed between patients with genotype 1b and a high viral load showing substitution of amino acid 70 in the core region of HCV ( mutant , n = 20 ) and patients without this substitution ( wild , n = 23 ) . PATIENTS AND METHODS : Forty-three CHC patients were studied , and received pegylated IFN plus ribavirin . P17181 -2 expression by Mo was determined using flow cytometry to measure the mean fluorescence intensity ( MFI ) before and up to 28 days after starting therapy . RESULTS : The EVR rate of the mutant group was significantly lower than that of the wild group ( 35 vs.70 % ) . The MFI of Mo was significantly higher in the wild group than in the mutant group before and also 3 , 7 , and 28 days after starting therapy . CONCLUSIONS : Mutation of HCV was related to lower P17181 -2 expression by Mo before and after starting therapy . P37231 ligands are potent inhibitors of angiogenesis in vitro and in vivo . P37231 ( PPARgamma ) is a nuclear receptor that functions as a transcription factor to mediate ligand-dependent transcriptional regulation . Activation of PPARgamma by the naturally occurring ligand , 15-deoxy-Delta12,14-prostaglandin J2 ( 15d-PGJ2 ) , or members of a new class of oral antidiabetic agents , e.g. BRL49653 and ciglitizone , has been linked to adipocyte differentiation , regulation of glucose homeostasis , inhibition of macrophage and monocyte activation , and inhibition of tumor cell proliferation . Here we report that human umbilical vein endothelial cells ( HUVEC ) express PPARgamma mRNA and protein . Activation of PPARgamma by the specific ligands 15d-PGJ2 , BRL49653 , or ciglitizone , dose dependently suppresses HUVEC differentiation into tube-like structures in three-dimensional collagen gels . In contrast , specific PPARalpha and -beta ligands do not affect tube formation although mRNA for these receptors are expressed in HUVEC . PPARgamma ligands also inhibit the proliferative response of HUVEC to exogenous growth factors . Treatment of HUVEC with 15d-PGJ2 also reduced mRNA levels of vascular endothelial cell growth factor receptors 1 ( Flt-1 ) and 2 ( Flk/ P35968 ) and urokinase plasminogen activator and increased plasminogen activator inhibitor-1 ( P05121 ) mRNA . Finally , administration of 15d-PGJ2 inhibited vascular endothelial cell growth factor-induced angiogenesis in the rat cornea . These observations demonstrate that PPARgamma ligands are potent inhibitors of angiogenesis in vitro and in vivo , and suggest that PPARgamma may be an important molecular target for the development of small-molecule inhibitors of angiogenesis . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . Genetic aspects of ischemic stroke : coagulation , homocysteine , and lipoprotein metabolism as potential risk factors . Stroke is one of the most common causes of death and long term disability throughout the world . It may be the outcome of a number of monogenic disorders or , more commonly , a polygenic multifactorial disease . Numerous studies have investigated the role of genetics in the pathogenesis of ischemic stroke , with varied and often contradictory results . The candidate ' stroke risk ' genes affecting haemostasis ( P12259 , F2 , P02671 / P02675 , P08709 , P00488 , P04275 , P00748 , P05121 , P05106 / P08514 , P17301 , P07359 , TPA , Q96IY4 , P07204 , PZ , P08758 ) , homocysteine metabolism ( P42898 , P35520 , Q99707 ) , and lipid metabolism ( apo E , P06858 , P11597 , O95477 , apo AI , apo CIII , apo AIV , apo AV , apo B , apo H , apo(a) , P27169 /2/3 , P01130 / P78380 ) are evaluated in this review . By examining meta-analyses and case-control studies , we made a classification of gene/gene polymorphisms according to the degree of association with ischemic stroke risk . The data assembled could be very useful for further meta-analysis and for future clinical applications . An acetylcholinesterase inhibitor , eserine , induces long-term depression at P07451 - P00915 synapses in the hippocampus of adult rats . Studies in humans and rodents support a role for muscarinic ACh receptor ( mAChR ) and nicotinic AChR in learning and memory , and both regulate hippocampal synaptic plasticity using complex and often times opposing mechanisms . P22303 ( P22303 ) inhibitors are commonly prescribed to enhance cholinergic signaling in Alzheimer 's disease in hopes of rescuing cognitive function , caused , in part , by degeneration of cholinergic innervation to the hippocampus and cortex . Unfortunately , therapeutic efficacy is moderate and inconsistent , perhaps due to unanticipated mechanisms . M1 mAChRs bidirectionally control synaptic strength at P07451 - P00915 synapses ; weak pharmacological activation using carbachol ( CCh ) facilitates potentiation , whereas strong agonism induces muscarinic long-term depression ( mLTD ) via an P29323 -dependent mechanism . Here , we tested the prediction that accumulation of extracellular ACh via inhibition of P22303 is sufficient to induce LTD at P07451 - P00915 synapses in hippocampal slices from adult rats . Although P22303 inhibition with eserine induces LTD , it unexpectedly does not share properties with mLTD induced by CCh , as reported previously . DB00981 -LTD was prevented by the M3 mAChR-preferring antagonist 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide ( 4-DAMP ) , and pharmacological inhibition of MEK was completely ineffective . Additionally , pharmacological inhibition of p38 MAPK prevents mLTD but has no effect on eserine-LTD . Finally , long-term expression of eserine-LTD is partially dependent on a decrease in presynaptic release probability , likely caused by tonic activation of mAChRs by the sustained increase in extracellular ACh . Thus these findings extend current literature by showing that pharmacological P22303 inhibition causes a prolonged decrease in presynaptic glutamate release at P07451 - P00915 synapses , in addition to inducing a likely postsynaptic form of LTD . [ The chemical structure and pharmacological properties of a novel isoxazolidinedione insulin sensitizer , DB04971 ] . DB04971 is an isoxazolidine-3,5-dione derivative . This drug activates both Q07869 gamma and Q07869 alpha , and shows not only a hypoglycemic effect but also a stronger triglyceride-lowering effect than the thiazolidine-2,4-diones . DB04971 improved both the impaired insulin-stimulated autophosphorylation levels of Zucker fatty rats and impaired insulin-induced P14672 translocation to the plasma membrane as well as insulin-induced glucose uptake in high fat diet rats , indicating that DB04971 enhances insulin signaling and reduces insulin resistance . Furthermore , DB04971 prevented several diabetic complications , such as cataract , nephropathy , and neuropathy in Zucker diabetic fatty rats . As a non-thiazolidinedione insulin sensitizer , DB04971 has been the first to start clinical trials and is currently undergoing evaluation in clinical studies for diabetic patients . Gene expression profiles for predicting the efficacy of the anticancer drug 5-fluorouracil in breast cancer . Chemotherapy is an important postsurgery adjuvant therapy in the treatment of breast cancer . However , because of the individual genotype differences of patients , the drug efficacy differs from person to person , even when the same chemotherapy drug is administered . The purpose of this research was to probe the gene expression profiles to predict the efficacy of 5-fluorouracil ( DB00544 ) , the common drug used in chemotherapy for various type of cancers , in Taiwanese breast cancer patients . Microarray analysis was conducted on the cancer cell line ZR-75-1 with and without DB00544 stimulation to identify the differentially expressed genes . The significant overexpressed gene groups were selected after bioinformatics software analysis to explore the molecular mechanism of DB00544 . Six strains of breast cancer cell line purchased from American Type Culture Collection were used to analyze the expression profiles of the above target gene groups . Q14116 , Q9NRJ3 , P19875 , P00441 , P01112 , P22570 , and P36222 genes were significantly differentially expressed in DB00544 responder and nonresponder cell lines . The selected gene groups were validated with 20 strains of breast cancer primary cultures established previously in our laboratory . The experimental results demonstrated that Q96IP4 , Q14116 , Q9NRJ3 , P01375 , P19875 , Q96JA3 , P01112 , P22570 , and P36222 genes showed statistically significant differential expression between primary breast cancer culture cells that respond and nonrespond to DB00544 . Six genes , Q14116 , Q9NRJ3 , P19875 , P01112 , P22570 , and P36222 , showed significant differential expression pattern in both American Type Culture Collection and primary breast cancer cultured cells . The findings of this study may serve as basis for predicting the effectiveness of DB00544 on breast cancer . Msx2 promotes cardiovascular calcification by activating paracrine Wnt signals . In diabetic P01130 -/- mice , an ectopic P12643 -Msx2 gene regulatory program is upregulated in association with vascular calcification . We verified the procalcific actions of aortic Msx2 expression in vivo . CMV-Msx2 transgenic ( CMV-Msx2Tg(+) ) mice expressed 3-fold higher levels of aortic Msx2 than nontransgenic littermates . On high-fat diets , CMV-Msx2Tg(+) mice exhibited marked cardiovascular calcification involving aortic and coronary tunica media . This corresponded to regions of Msx2 immunoreactivity in adjacent adventitial myofibroblasts , suggesting a potential paracrine osteogenic signal . To better understand Msx2-regulated calcification , we studied actions in 10T1/2 cells . We found that conditioned media from Msx2-transduced 10T1/2 cells ( Msx2-CM ) is both pro-osteogenic and adipostatic ; these features are characteristic of Wnt signaling . Msx2-CM stimulated Wnt-dependent TCF/LEF transcription , and Msx2-transduced cells exhibited increased nuclear beta-catenin localization with concomitant alkaline phosphatase induction . Msx2 upregulated Wnt3a and Wnt7a but downregulated expression of the canonical inhibitor Dkk1 . Dkk1 treatment reversed osteogenic and adipostatic actions of Msx2 . DB06285 , a Q03431 agonist that inhibits murine vascular calcification , suppressed vascular P12643 -Msx2-Wnt signaling . Analyses of CMV-Msx2Tg(+) mice confirmed that Msx2 suppresses aortic Dkk1 and upregulates vascular Wnts ; moreover , TOPGAL(+) ( Wnt reporter ) ; CMV-Msx2Tg(+) mice exhibited augmented aortic LacZ expression . Thus , Msx2-expressing cells elaborated an osteogenic milieu that promotes vascular calcification in part via paracrine Wnt signals . Genetic variations in humans associated with differences in the course of hepatitis C . The outcome of hepatitis C virus ( HCV ) infection varies among individuals , but the genetic factors involved remain unknown . We conducted a population-based association study in which 238 Japanese individuals positive for anti-HCV antibody were genotyped for 269 single nucleotide polymorphisms ( SNPs ) in 103 candidate genes that might influence the course of infection . Altogether , 50 SNPs in 32 genes were listed . Genetic polymorphisms in P05112 , P25025 , Q13651 , PRL , P00813 , P19838 , O75791 , Q9Y6J0 , P48551 , P40305 , IFI41 , P19438 , P05062 , Q10567 , O00204 , P01133 , P00533 , P01137 , Q14767 , and P01730 were associated with persistent viremia ( P < 0.05 ) , whereas those in P01584 , Q01638 , P14784 , P42701 , Q13478 , P42229 , O75791 , Q9Y6J0 , P17181 , Mx1 , P34820 , Q08830 , Q14767 , P28906 , and P33681 were associated with different serum alanine aminotransferase levels in HCV carriers ( P < 0.05 ) . The sorted genes allow us to draw novel hypotheses for future studies of HCV infection to ultimately identify bona fide genes and their variations . Crystal structures of adrenodoxin reductase in complex with NADP+ and NADPH suggesting a mechanism for the electron transfer of an enzyme family . P22570 is a flavoenzyme that shuffles electrons for the biosynthesis of steroids . Its chain topology belongs to the glutathione reductase family of disulfide oxidoreductases , all of which bind DB03147 at equivalent positions . The three reported structures of adrenodoxin reductase were ligated with reduced and oxidized NADP and have now confirmed this equivalence also for the NADP-binding site . Remarkably , the conformations and relative positions of the prosthetic group DB03147 and the cofactor NADP have been conserved during protein evolution despite very substantial changes in the polypeptide . The ligated enzymes showed small changes in the domain positions . When compared with the structure of the NADP-free enzyme , these positions correspond to several states of the domain motion during NADP binding . On the basis of the observed structures , we suggest an enzymatic mechanism for the subdivision of the received two-electron package into the two single electrons transferred to the carrier protein adrenodoxin . The data banks contain 10 sequences that are closely related to bovine adrenodoxin reductase . Most of them code for gene products with unknown functions . Within this family , the crucial residues of adrenodoxin reductase are strictly conserved . Moreover , the putative docking site of the carrier is rather well conserved . Five of the family members were assigned names related to ferredoxin:NADP(+) reductase , presumably because adrenodoxin reductase was considered a member of this functionally similar family . Since this is not the case , the data bank entries should be corrected . The regulation of rotenone-induced inflammatory factor production by DB00171 -sensitive potassium channel expressed in BV-2 cells . Our previous studies have demonstrated that activating DB00171 -sensitive potassium channel ( K( DB00171 ) channel ) , not only improved Parkinsonian behavior and neurochemical symptoms , but also reduced P35228 activity and mRNA levels in striatum and nigra of rotenone rat model of Parkinson 's disease ( PD ) . In this study , it was first shown that the subunits of K( DB00171 ) channels are expressed in BV-2 cells , and then it was investigated whether K( DB00171 ) channel was involved in regulating inflammatory factor production from BV-2 cells activated by rotenone . It was found that K( DB00171 ) channel was expressed in BV-2 cell and formed by the combination of Kir 6.1 and Q09428 2A/2B . K( DB00171 ) channel openers ( KCOs ) including pinacidil , diazoxide and iptakalim ( Ipt ) exerted beneficial effects on rotenone-induced morphological alterations of BV-2 cells , decreased tumor necrosis factor alpha ( P01375 ) production and the expression and activity of inducible isoform of nitric oxide synthase ( P35228 ) . Either glibenclamide or 5-hydroxydecanoate acid ( a selective mitochondrial K( DB00171 ) channel blocker ) could abolish the effects of KCOs , suggesting that K( DB00171 ) channels , especially mitochondrial DB00171 -sensitive potassium channels ( mitoK( DB00171 ) channels ) , played a crucial role in preventing the activation of BV-2 cells , and subsequently the production of a variety of proinflammatory factors . Therefore , activation of K( DB00171 ) channel might be a new therapeutic strategy for treating neuroinflammatory and neurodegenerative disorders . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . Regulatory regions of growth-related genes can activate an exogenous gene of the alpha-fetoprotein promoter to a comparable degree in human hepatocellular carcinoma cells . We examined the transcriptional activation by the regulatory regions of the midkine ( MK ) , survivin ( Q09428 ) , cyclooxygenase-2 ( P35354 ) , telomerase reverse transcriptase ( O14746 ) and alpha-fetoprotein ( AFP ) genes in human hepatocellular carcinoma cells . Luciferase assays showed that the Q09428 regulatory region exhibited the greatest activity and that the MK regulatory region activated the reporter gene better than the enhancer-linked AFP promoter even in high-AFP-producing cells . The P35354 and O14746 regulatory regions also activated the reporter gene better than the AFP enhancer/promoter in intermediate-AFP-producing cells . Combination of the regulatory regions arranged in tandem modulated their transcriptional activities , depending on the arrangement of the promoters and cells examined . These data suggested that the regulatory regions of the growth-related genes could be useful to activate a therapeutic gene in hepatocellular carcinoma cells irrespective of the amounts of AFP production but combinatory use of the promoter regions could not always contribute to enhanced activity . Endothelial cells in co-culture enhance embryonic stem cell differentiation to pancreatic progenitors and insulin-producing cells through BMP signaling . Endothelial cells ( ECs ) represent the major component of the embryonic pancreatic niche and play a key role in the differentiation of insulin-producing β cells in vivo . However , it is unknown if ECs promote such differentiation in vitro . We investigated whether interaction of ECs with mouse embryoid bodies ( EBs ) in culture promotes differentiation of pancreatic progenitors and insulin-producing cells and the mechanisms involved . We developed a co-culture system of mouse EBs and human microvascular ECs ( HMECs ) . An increase in the expression of the pancreatic markers P52945 , Ngn3 , Nkx6.1 , proinsulin , P11168 , and Ptf1a was observed at the interface between EBs and ECs ( EB-EC ) . No expression of these markers was found at the periphery of EBs cultured without ECs or those co-cultured with mouse embryonic fibroblasts ( MEFs ) . At EB-EC interface , proinsulin and Nkx6.1 positive cells co-expressed phospho- Q15797 /5/8 ( pSmad1/5/8 ) . Therefore , EBs were treated with HMEC conditioned media ( HMEC-CM ) suspecting soluble factors involved in bone morphogenetic protein ( BMP ) pathway activation . Upregulation of P52945 , Ngn3 , Nkx6.1 , insulin-1 , insulin-2 , amylin , Q09428 , GKS , and amylase as well as down-regulation of P61278 were detected in treated EBs . In addition , higher expression of P12643 /-4 and their receptor ( P36894 ) were also found in these EBs . Recombinant human P12643 ( rhBMP-2 ) mimicked the effects of the HMEC-CM on EBs . Q13253 ( Q13253 ) , a BMP antagonist , partially inhibited these effects . These results indicate that the differentiation of EBs to pancreatic progenitors and insulin-producing cells can be enhanced by ECs in vitro and that BMP pathway activation is central to this process . Differences in transcript levels of ABC transporters between pancreatic adenocarcinoma and nonneoplastic tissues . OBJECTIVES : The aim of this study was to evaluate transcript levels of all 49 human DB00171 -binding cassette transporters ( ABCs ) in one of the most drug-resistant cancers , namely , the pancreatic ductal adenocarcinoma ( PDAC ) . Association of ABCs levels with clinical-pathologic characteristics and P01116 mutation status was followed as well . METHODS : Tumors and adjacent nonneoplastic tissues were obtained from 32 histologically verified PDAC patients . The transcript profile of ABCs was assessed using quantitative real-time polymerase chain reaction with a relative standard curve . P01116 mutations in exon 2 were assessed by high-resolution melting analysis and sequencing . RESULTS : Most ABCs were deregulated in PDAC and 10 ABCs were associated with clinical-pathologic characteristics . P01116 mutations did not change the global expression profile of ABCs . CONCLUSIONS : The expression of ABC transporters was significantly deregulated in PDAC tumors when compared to nonmalignant tissues . The observed up-regulation of P21439 , O95342 , P33527 , O15438 , O15440 , Q5T3U5 , and Q9UNQ0 in tumors may contribute to the generally poor treatment response of PDAC . The up-regulation of O95477 , Q8IZY2 , and P45844 implicates a serious impairment of cellular cholesterol homeostasis in PDAC . On the other hand , the observed down-regulation of Q99758 , O95255 , P13569 , and Q09428 suggests a possible role of stem cells in the development and progression of PDAC . Green tea polyphenols in the prevention of colon cancer . Several plant-based nutrients and non-nutrients that can inhibit mutagenesis and proliferation have been identified . Some of the most promising nutrients identified as chemopreventive agents in colon cancer prevention include isoflavones , curcumin , calcium , vitamin D and more recently Green tea polyphenols ( GTP ) . In addition to inhibiting mutagenesis and proliferation , these compounds are relatively non-toxic , are of low cost and can be taken orally or as a part of the daily diet . Epidemiological and laboratory studies have identified epigallocatechin gallate ( EGCG ) in green tea polyphenols ( GTP ) , as the most potent chemopreventive agent that can induce apoptosis , suppress the formation and growth of human cancers including colorectal cancers ( CRC ) . It is only logical then , that future clinical studies should focus on examining the efficacy of phytochemicals such as EGCG in cancer chemoprevention as an alternative to pharmacological agents , especially in populations where administration of P35354 inhibitors , DB00945 and NSAIDS is contraindicated . The goal of this review is to provide the rationale , and discuss the use of EGCG in GTP as a chemopreventive agent for prevention of colon carcinogenesis and present evidence for the efficacy and safety of these agents based on epidemiological , animal , in vitro studies and Phase I clinical trials . Targeted disruption of the methionine synthase gene in mice . Alterations in homocysteine , methionine , folate , and/or B12 homeostasis have been associated with neural tube defects , cardiovascular disease , and cancer . Q99707 , one of only two mammalian enzymes known to require vitamin B12 as a cofactor , lies at the intersection of these metabolic pathways . This enzyme catalyzes the transfer of a methyl group from 5-methyl- DB00116 to homocysteine , generating DB00116 and methionine . Human patients with methionine synthase deficiency exhibit homocysteinemia , homocysteinuria , and hypomethioninemia . They suffer from megaloblastic anemia with or without some degree of neural dysfunction and mental retardation . To better study the pathophysiology of methionine synthase deficiency , we utilized gene-targeting technology to inactivate the methionine synthase gene in mice . On average , heterozygous knockout mice from an outbred background have slightly elevated plasma homocysteine and methionine compared to wild-type mice but seem to be otherwise indistinguishable . Homozygous knockout embryos survive through implantation but die soon thereafter . Nutritional supplementation during pregnancy was unable to rescue embryos that were completely deficient in methionine synthase . Whether any human patients with methionine synthase deficiency have a complete absence of enzyme activity is unclear . These results demonstrate the importance of this enzyme for early development in mice and suggest either that methionine synthase-deficient patients have residual methionine synthase activity or that humans have a compensatory mechanism that is absent in mice . Effect of P35354 inhibitor lumiracoxib and the P01375 -α antagonist etanercept on TNBS-induced colitis in Wistar rats . Crohn 's disease ( CD ) is associated with gut barrier dysfunction . Besides the baseline barrier defect , a subgroup of patients also expresses an intestinal barrier hyperresponsiveness to nonsteroidal anti-inflammatory drugs . On the other hand , the anti-tumour necrosis factor alpha ( P01375 -α ) treatment has brought benefits to these patients . Thus , this study aimed to evaluate the effect of lumiracoxib , a selective-cyclooxygenase-2 ( P35354 ) inhibitor , and DB00005 ( ETC ) , a P01375 -α antagonist on the 2,4,6-trinitrobenzene sulfonic acid ( TNBS ) -induced experimental colitis . A total of 47 Wistar rats were randomized into seven groups , as follows : ( 1 ) Sham : sham induced-colitis ; ( 2 ) TNBS : nontreated induced-colitis ; ( 3 ) DB01283 control ; ( 4 ) DB01283 -treated induced-colitis ; ( 5 ) ETC control ; ( 6 ) ETC-treated induced-colitis ; ( 7 ) DB01283 -ETC-treated induced-colitis . Rats from groups 6 and 7 presented significant improvement of macroscopic and histopathological damages in the distal colon . The gene expression of P35354 mRNA , as well of P01375 -α mRNA , decreased significantly in groups 6 and 7 compared to the TNBS nontreated and lumiracoxib-treated groups . The treatment only with lumiracoxib did not reduce the inflammation on TNBS-induced experimental colitis . ETC attenuated the damage seen in the colon and reduced the inflammation caused by TNBS . Our results suggest that down-regulation of P01375 -α and P35354 resulted in a decrease in inflammation caused by TNBS and thus provided some protection from the colonic damage caused by TNBS .

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Anti- P15692 therapy for the treatment of glaucoma : a focus on ranibizumab and bevacizumab . INTRODUCTION : Anti- P15692 therapy has been widely used in the treatment of ocular neovascular diseases . Because of their anti-angiogenic and anti-fibrotic properties , anti- P15692 antibodies such as bevacizumab and ranibizumab have emerged as an adjunctive treatment modality in glaucoma to improve success of conventional treatments . AREAS COVERED : DB01270 is an anti- P15692 antigen binding fragment currently indicated in neovascular age-related macular degeneration as well as macular edema following retinal vein occlusion . Several off-label uses include the treatment of neovascular glaucoma to regress/suppress iris and iridocorneal angle neovascularization and the modulation of wound healing after glaucoma filtration surgery . DB00112 is a full-length anti- P15692 antibody , which is also being used in aforementioned eye conditions off-label . An overview of these anti- P15692 antibodies and the results of preclinical and clinical studies regarding their use in the treatment of glaucoma are presented . EXPERT OPINION : Early studies on the utility of both bevacizumab and ranibizumab in neovascular glaucoma and filtration surgery reported promising results . However , a large-scale randomized clinical trial as well as comparative studies between the two anti- P15692 antibodies are currently lacking . A single dose of ranibizumab costs approximately 40 times as much as a single dose of bevacizumab . Clinicians should take this into account , in addition to their differences in the efficacy and safety , when treating patients . Modification of the reactivity of three amino-acid residues in elongation factor 2 during its binding to ribosomes and translocation . The accessibility of three amino acids of P13639 , located within highly conserved regions near the N- and C-terminal extremities of the molecule ( the E region and the DB02059 region , respectively ) to modifying enzymes has been compared within nucleotide-complexed P13639 and ribosomal complexes that mimic the pre- and posttranslocational ones : the high-affinity complex ( P13639 ) -nonhydrolysable GTP analog GuoPP[CH2]P ribosome and the low-affinity ( P13639 ) -GDP-ribosome complex , P13639 and ribosomes being from rat liver . We studied the reactivity of two highly conserved residues diphthamide-715 and DB00125 -66 , to diphtheria-toxin-dependent ADP-ribosylation and trypsin attack , and of a threonine that probably lies between residues 51 and 60 , to phosphorylation by a Ca2+/calmodulin-dependent protein kinase . DB03223 715 and this threonine residue were unreactive within the high-affinity complex but seemed fully reactive in the low-affinity complex . DB00125 -66 was resistant to trypsin in both complexes . The possible involvement of the E and DB02059 regions of P13639 in the interaction with ribosome in the two complexes is discussed . Management of ocular inflammation and pain following cataract surgery : focus on bromfenac ophthalmic solution . Recently , several new ophthalmic NSAID products have been introduced for commercial use in the United States . The purpose of this review is to briefly overview the ophthalmic NSAIDs currently in use and to discuss the management of postoperative ocular inflammation and pain following cataract surgery with a particular focus on bromfenac ophthalmic solution 0.09 % . DB00963 ophthalmic solution 0.09 % is indicated for the reduction of ocular pain and inflammation following cataract surgery . Studies have shown that bromfenac ophthalmic solution 0.09 % has equivalent efficacy to the other topical NSAIDs in reducing postsurgical inflammation and controlling pain . The unique chemical structure of bromfenac makes it both a potent inhibitor of the P35354 enzyme and a highly lipophilic molecule that rapidly penetrates to produce early and sustained drug levels in all ocular tissues . Clinically , these pharmacokinetic features are manifested in a rapid reduction of postsurgical inflammation and pain with bid dosing . DB00963 ophthalmic solution 0.09 % is a versatile agent and is effective when used as either monotherapy or as an adjunct therapy to steroids . Role of serotonin in the regulation of interferon-gamma production by human natural killer cells . Monocytes , recovered directly from peripheral blood by counter-current centrifugal elutriation ( CCE ) , were shown to provide two regulatory signals for induction of interferon-gamma ( P01579 ) in natural killer ( NK ) cells in response to interleukin-2 ( P60568 ) : an upregulating signal and an inhibitory signal . The inhibitory signal was time-dependent , irreversible , and operating on a pretranslational level , as indicated by the inability of enriched NK cells to accumulate P01579 mRNA in the presence of elutriated monocytes . Monocyte-induced inhibition of P01579 production was abrogated by the biogenic amine serotonin , acting via the 5-hydroxytryptamine , or serotonin ( P08908 ) , subset of serotonin receptors ( 5-HTR ) . Thereby , serotonin effectively promoted P01579 production in the presence of monocytes . We conclude that serotonergic P08908 receptors transduce signals that are required for NK cells to produce P01579 in response to P60568 . Phosphorylation at serine 208 of the 1alpha,25-dihydroxy P11473 modulates the interaction with transcriptional coactivators . Upon ligand binding the 1alpha,25-dihydroxy P11473 ( P11473 ) undergoes a conformational change that allows interaction with coactivator proteins including P52701 / P12931 family members and the multimeric DRIP complex through the Q15648 subunit . Casein kinase II ( CKII ) phosphorylates P11473 both in vitro and in vivo at serine 208 within the hinge domain . This phosphorylation does not affect the ability of P11473 to bind DNA , but increases its ability to transactivate target promoters . Here , we have analyzed whether phosphorylation of P11473 by CKII modulates the ability of P11473 to interact with coactivators in vitro . We find that both mutation of serine 208 to aspartic acid ( VDRS208D ) or phosphorylation of P11473 by CKII enhance the interaction of P11473 with Q15648 in the presence of 1alpha,25-dihydroxy DB00169 . We also find that the mutation VDRS208D neither affects the ability of this protein to bind DNA nor to interact with Q15788 and RXRalpha . Together , our results indicate that phosphorylation of P11473 at serine 208 contributes to modulate the affinity of P11473 for the DRIP complex and therefore may have a role in vivo regulating P11473 -mediated transcriptional enhancement . [ Treatment of neovascular age-related macular degeneration with DB01270 /Lucentis ] . Vascular endothelial growth factor ( P15692 ) is considered to play an essential role in the pathogenesis of age-related macular degeneration due to its vascular permeability-inducing and angiogenic properties . DB01270 , a small antibody fragment designed to competitively bind all P15692 isoforms , passes after intravitreal injection all retinal layers reaching the retinal pigment epithelium-choroid complex . Experimental animal models showed the drug to be safe and effective . Subsequently , Phase I/II clinical trials conducted in patients with neovascular AMD demonstrated a good safety profile , and a significant functional benefit . DB01270 therapy repeated every four weeks for the treatment of neovascular AMD is currently in Phase III clinical trials . Combination therapy trials aiming for improved treatment durability and effectiveness are currently ongoing as well as new treatment strategies using intermittent , optical coherence tomography ( O75051 ) guided therapy . Anti- P15692 therapy using DB01270 is a promising new treatment option for neovascular AMD . Ocular pharmacokinetics of a single dose of bromfenac sodium ophthalmic solution 0.1 % in human aqueous humor . PURPOSE : The aim of this study was to evaluate the ocular pharmacokinetics of a single dose of bromfenac sodium ophthalmic solution 0.1 % in subjects undergoing routine cataract surgery with intraocular lens implantation . METHODS : An open-label , phase II confirmatory study of 54 subjects undergoing cataract surgery with intraocular lens implantation . A single drop of bromfenac sodium ophthalmic solution 0.1 % was administered at 30 , 60 , 90 , 120 , 180 , or 240 min prior to the initiation of cataract surgery . Samples of aqueous humor were aspirated through a paracentesis and analyzed by using high-performance liquid chromatography . Based upon these data , predicted concentrations of bromfenac in the aqueous humor over 24 h were generated by using computer simulation and compared with the IC(50) for bromfenac as a measure of anti-inflammatory efficacy . RESULTS : Peak aqueous-humor concentrations of bromfenac occurred between 150 and 180 min following ophthalmic dosing , with a mean concentration of 78.7 ng/mL . The level of bromfenac decreased in a log-linear fashion with an elimination-rate constant of 1.4 . DB00963 remained above the IC(50) value of cyclo-oxygenase-2 ( P35354 ) during the evaluated time points and over the 12-h dosing interval , using a computer model of extrapolated time points and assuming first-order elimination . CONCLUSIONS : Pharmacokinetic modeling , based upon samples collected over 240 min after a single dose of bromfenac sodium ophthalmic solution 0.1 % suggests that aqueous-humor concentrations remain at clinically effective levels ( above its IC(50) value for P35354 ) for over 12 h . Based upon this rationale , these results supported clinical trials that demonstrated the efficacy of twice-daily dosing of bromfenac sodium ophthalmic solution 0.1 % to manage patients with postoperative ocular pain and inflammation . DB03615 inhibits the chaperone activity of protein disulfide isomerase . In the process of screening of proteins binding to ribostamycin in bovine liver using the affinity column chromatography , we found that ribostamycin inhibited the chaperone activity of protein disulfide isomerase ( P07237 ) , but it did not inhibit the isomerase activity . P07237 was identified by SDS-PAGE , Western blotting , and N-terminal amino acid sequence analysis . A 100:1 molar ratio of ribostamycin to P07237 was almost sufficient to completely inhibit the chaperone activity of P07237 . The binding affinity of ribostamycin to purified bovine P07237 was determined by the Biacore system , which gave a K(D) value of 3.19 x 10(-4) M . This suggests that ribostamycin binds to region distinct from the CGHC motif of P07237 . This is the first report to describe the inhibitor of the chaperone activity of P07237 . Chronic constipation : let symptom type and severity direct treatment . Increased fiber intake through diet or fiber supplements is an appropriate initial therapy for chronic constipation . Osmotic and stimulant laxatives may be administered to patients who do not respond to more conservative measures if the limitations of these agents are explained . DB01079 , a selective 5-hydroxytryptamine type 4 ( Q13639 ) receptor partial agonist , is more effective than placebo at relieving symptoms of chronic idiopathic constipation in patients younger than 65 years of age . Patients with suspected defecation disorders and those with treatment-refractory symptoms should be referred to a gastroenterologist for further evaluation . Autoantibodies against four kinds of neurotransmitter receptors in psychiatric disorders . There is a hypothesis that autoimmune abnormalities in neurotransmitter receptors might cause some psychiatric disorders . Using a sensitive radioligand assay , we detected serum autoantibodies to recombinant human muscarinic cholinergic receptor 1 ( P11229 , 34.4 % ) , mu-opioid receptor ( P35372 , 13.1 % ) , P08908 ( P08908 , 7.4 % ) , and dopamine receptor D2 ( P14416 , 4.9 % ) in 122 psychiatric patients . Positive antibodies to P11229 were found in 34.1 % , 34.9 % , 33.3 % , and 9.1 % of patients with schizophrenic disorders ( n=44 ) , mood disorders ( n=63 ) , other psychiatric disorders ( n=15 ) and autoimmune diseases ( n=33 ) , respectively . All three patients with neuroleptic maliganant syndrome had high activities of autoantibodies to P11229 , P35372 , and/or P08908 . Our data suggest that autoimmunity to neurotransmitter receptors might be associated with the induction of psychiatric symptoms and have some relation to neuroleptic malignant syndrome . Porous polyimide membranes prepared by wet phase inversion for use in low dielectric applications . A wet phase inversion process of polyamic acid ( PAA ) allowed fabrication of a porous membrane of polyimide ( PI ) with the combination of a low dielectric constant ( 1.7 ) and reasonable mechanical properties ( Tensile strain : 8.04 % , toughness : 3.4 MJ/m3 , tensile stress : 39.17 MPa , and young modulus : 1.13 GPa ) , with further thermal imidization process of PAA . PAA was simply synthesized from purified pyromellitic dianhydride ( PMDA ) and 4,4-oxydianiline ( ODA ) in two different reaction solvents such as γ- DB04699 ( Q9BVC4 ) and N-methyl-2-pyrrolidinone ( NMP ) , which produce Mw/ P07237 of 630,000/1.45 and 280,000/2.0 , respectively . The porous PAA membrane was fabricated by the wet phase inversion process based on a solvent/non-solvent system via tailored composition between Q9BVC4 and NMP . The porosity of PI , indicative of a low electric constant , decreased with increasing concentration of Q9BVC4 , which was caused by sponge-like formation . However , due to interplay between the low electric constant ( structural formation ) and the mechanical properties , Q9BVC4 was employed for further exploration , using toluene and acetone vs. DI-water as a coagulation media . Non-solvents influenced determination of the PAA membrane size and porosity . With this approach , insight into the interplay between dielectric properties and mechanical properties will inform a wide range of potential low-k material applications . DB01819 cycling via mitochondrial phosphoenolpyruvate carboxykinase links anaplerosis and mitochondrial GTP with insulin secretion . Pancreatic beta-cells couple the oxidation of glucose to the secretion of insulin . Apart from the canonical K( DB00171 )-dependent glucose-stimulated insulin secretion ( GSIS ) , there are important K( DB00171 )-independent mechanisms involving both anaplerosis and mitochondrial GTP ( mtGTP ) . How mtGTP that is trapped within the mitochondrial matrix regulates the cytosolic calcium increases that drive GSIS remains a mystery . Here we have investigated whether the mitochondrial isoform of phosphoenolpyruvate carboxykinase ( Q16822 ) is the GTPase linking hydrolysis of mtGTP made by succinyl- DB01992 synthetase ( SCS-GTP ) to an anaplerotic pathway producing phosphoenolpyruvate ( PEP ) . Although cytosolic PEPCK ( P35558 ) is absent , Q16822 message and protein were detected in P01308 -1 832/13 cells , rat islets , and mouse islets . PEPCK enzymatic activity is half that of primary hepatocytes and is localized exclusively to the mitochondria . Novel (13)C-labeling strategies in P01308 -1 832/13 cells and islets measured substantial contribution of Q16822 to the synthesis of PEP . As high as 30 % of PEP in P01308 -1 832/13 cells and 41 % of PEP in rat islets came from Q16822 . The contribution of Q16822 to overall PEP synthesis more than tripled with glucose stimulation . Silencing the Q16822 gene completely inhibited GSIS underscoring its central role in mitochondrial metabolism-mediated insulin secretion . Given that mtGTP synthesized by SCS-GTP is an indicator of TCA flux that is crucial for GSIS , Q16822 is a strong candidate to link mtGTP synthesis with insulin release through anaplerotic PEP cycling . Neurological impairment in experimental antiphospholipid syndrome is associated with increased ligand binding to hippocampal and cortical serotonergic P08908 receptors . The antiphospholipid syndrome ( APS ) is an autoimmune disease where the presence of high titers of circulating autoantibodies causes thrombosis with consecutive infarcts . In experimental APS ( eAPS ) , a mouse model of APS , behavioral abnormalities develop in the absence of vessel occlusion or infarcts . Using brain hemispheres of control and eAPS mice with documented neurological and cognitive deficits , we checked for lymphocytic infiltration , activation of glia and macrophages , as well as alterations of ligand binding densities of various neurotransmitter receptors to unravel the molecular basis of this abnormal behavior . Lymphocytic infiltrates were immunohistochemically characterized using antibodies against CD3 , P01730 , CD8 and forkhead box P09131 ( Foxp3 ) , respectively . P14136 , Iba1 and P34810 -immunohistochemistry was performed , to check for activation of astrocytes , microglia and macrophages . Ligand binding densities of DB01221 , AMPA , GABAA and P08908 receptors were analyzed by in vitro receptor autoradiography . No significant inflammatory reaction occurred in eAPS mice . There was neither activation of astrocytes or microglia nor accumulation of macrophages . Binding values of excitatory and inhibitory neurotransmitter receptors were largely unchanged . However , ligand binding densities of the modulatory serotonergic P08908 receptors in the hippocampus and in the primary somatosensory cortex of eAPS mice were significantly upregulated which is suggested to induce the behavioral abnormalities observed . Neuronal ablation of p-Akt at Ser473 leads to altered P08908 /2A receptor function . The serotonergic system regulates a wide range of behavior , including mood and impulsivity , and its dysregulation has been associated with mood disorders , autism spectrum disorder , and addiction . Diabetes is a risk factor for these conditions . P01308 resistance in the brain is specifically associated with susceptibility to psychostimulant abuse . Here , we examined whether phosphorylation of Akt , a key regulator of the insulin signaling pathway , controls serotonin ( 5-HT ) signaling . To explore how impairment in Akt function regulates 5-HT homeostasis , we used a brain-specific rictor knockout ( KO ) mouse model of impaired neuronal phosphorylation of Akt at Ser473 . Cortical P08908 and 5- Q13049 receptor binding was significantly elevated in rictor KO mice . Concomitant with this elevated receptor expression , the P08908 receptor agonist 8-Hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) led to an increased hypothermic response in rictor KO mice . The increased cortical P08908 receptor density was associated with higher P08908 receptor levels on the cortical cell surface . In contrast , rictor KO mice displayed significantly reduced head-twitch response ( HTR ) to the 5- Q13049 /C agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , with evidence of impaired 5- Q13049 /C receptor signaling . In vitro , pharmacological inhibition of Akt significantly increased P08908 receptor expression and attenuated DOI-induced 5- Q13049 receptor signaling , thereby lending credence to the observed in vivo cross-talk between neuronal Akt signaling and 5-HT receptor regulation . These data reveal that defective central Akt function alters 5-HT signaling as well as 5-HT-associated behaviors , demonstrating a novel role for Akt in maintaining neuronal 5-HT receptor function . Vascular endothelial growth factor signaling is required for the behavioral actions of antidepressant treatment : pharmacological and cellular characterization . This study extends earlier work on the role of vascular endothelial growth factor ( P15692 ) in the actions of antidepressant treatment in two key areas . First , by determining the requirement for P15692 in the actions of a 5-HT selective reuptake inhibitor ( SSRI ) , fluoxetine in behavioral models of depression/antidepressant response ; and second , by examining the role of the P08908 receptor subtype in the regulation of P15692 , and the cellular localization of antidepressant regulation of P15692 expression . The results show that pharmacological inhibition of P15692 receptor signaling blocks the behavioral actions of fluoxetine in rats subjected to chronic unpredictable stress . Infusions of SU5416 or SU1498 , two structurally dissimilar inhibitors of P15692 -Flk-1 receptor signaling , block the antidepressant effects of fluoxetine on sucrose preference , immobility in the forced swim test , and latency to feed in the novelty suppressed feeding paradigm . We also show that activation of P08908 receptors is sufficient to induce P15692 expression and that a P08908 antagonist blocks both the increase in P15692 and behavioral effects induced by fluoxetine . Finally , double labeling studies show that chronic fluoxetine administration increases P15692 expression in both neurons and endothelial cells in the hippocampus . Taken together these studies show that P15692 is necessary for the behavioral effects of the SSRI fluoxetine , as well as norepinephrine selective reuptake inhibitor , and that these effects may be mediated by P08908 receptors located on neurons and endothelial cells . DB00193 and another atypical opioid meperidine have exaggerated serotonin syndrome behavioural effects , but decreased analgesic effects , in genetically deficient serotonin transporter ( P31645 ) mice . The serotonin syndrome is a potential side-effect of serotonin-enhancing drugs , including antidepressants such as selective serotonin reuptake inhibitors ( SSRIs ) and monoamine oxidase inhibitors ( MAOIs ) . We recently reported a genetic mouse model for the serotonin syndrome , as serotonin transporter ( P31645 ) -deficient mice have exaggerated serotonin syndrome behavioural responses to the MAOI tranylcypromine and the serotonin precursor 5-hydroxy-l-tryptophan ( 5-HTP ) . As numerous case reports implicate the atypical opioids tramadol and meperidine in the development of the human serotonin syndrome , we examined tramadol and meperidine as possible causative drugs in the rodent model of the serotonin syndrome in P31645 wild-type ( +/+ ) , heterozygous ( +/- ) and knockout ( -/- ) mice . Comparisons were made with P31645 mice treated with either vehicle or morphine , an opioid not implicated in the serotonin syndrome in humans . Here we show that tramadol and meperidine , but not morphine , induce serotonin syndrome-like behaviours in mice , and we show that this response is exaggerated in mice lacking one or two copies of P31645 . The exaggerated response to tramadol in P31645 -/- mice was blocked by pretreatment with the P08908 antagonist WAY 100635 . Further , we show that morphine- , meperidine- and tramadol-induced analgesia is markedly decreased in P31645 -/- mice . These studies suggest that caution seems warranted in prescribing or not warning patients receiving SSRIs or MAOIs that dangerous side-effects may occur during concurrent use of tramadol and similar agents . These findings suggest that it is conceivable that there might be increased vulnerability in individuals with P31645 polymorphisms that may reduce P31645 by more than 50 % , the level in P31645 +/- mice . Endotoxin induced hyperlactatemia and hypoglycemia is linked to decreased mitochondrial phosphoenolpyruvate carboxykinase . AIMS : DB01819 carboxykinase ( PEPCK ) is the rate limiting enzyme for gluconeogenesis , and plays a key role in recycling lactate for glucose production . It is synthesized as two separate isoforms ; cytosolic ( P35558 , gene code ; P35558 ) and mitochondrial ( Q16822 , gene code ; Q16822 ) . Previous studies of gluconeogenesis in endotoxemia have focused solely on P35558 . We investigated the relative roles of the two isoforms in hepatic and renal gluconeogenesis in a rat model of endotoxic shock , and in cultured hepatocytes . MAIN METHODS : Rats were administered lipopolysaccharide ( 6 mg/kg ; LPS ) for 6 h . Cultured cells were incubated with lactate ( 5 mM ) with or without tumor necrosis factor alpha ( 1 - 10 ng/ml ) . Rat liver and kidney samples as well as cultured cells were subjected to subcellular fractionation to produce mitochondrial and cytosolic fractions for PEPCK activity assay . P35558 and Q16822 mRNA levels were measured using quantitative RT-PCR . KEY FINDINGS : In rat endotoxemia , hepatic Q16822 mRNA and Q16822 enzyme activity decreased by 53 % and 38 % , compared to sham controls . Hepatic P35558 mRNA levels increased by 44 % , but P35558 enzyme activity remained unchanged . The changes in hepatic Q16822 coincided with a marked hypoglycemia and hyperlactatemia as well as elevated plasma interleukin 1 beta ( IL1beta ) . Incubation of cultured hepatocytes with P01375 inhibited lactate-induced increases in glucose production , Q16822 mRNA levels and Q16822 enzyme activity but had no effect on P35558 mRNA levels or P35558 activity . SIGNIFICANCE : These results indicate that decreases in hepatic Q16822 play a key role in the manifestation of hyperlactatemia and hypoglycemia in endotoxemia . A novel 5-HT1-like receptor subtype mediates DB02527 synthesis in porcine pial vein . The 5-hydroxytryptamine ( 5-HT ) receptor subtype mediating 5-HT inhibition of spontaneous rhythmic contractions ( P12931 ) in the porcine pial vein was characterized . Results from pharmacological studies using in vitro tissue bath techniques indicated that the inhibitory effects of 5-HT on P12931 were qualitatively and quantitatively mimicked by 5-HT1-like agonists 5-methoxytryptamine ( 5-MT ) and 5-carboxamidotryptamine ( 5-CT ) . 5-HT- , 5-MT- , and 5-CT-induced inhibitions of P12931 were attenuated in a concentration-dependent manner by methysergide , which yielded similar pA2 values against these three agonists , suggesting that 5-HT , 5-MT , and 5-CT act on the same 5-HT1-like receptors . 5-MT inhibition of P12931 was not affected by blocking 5-HT2 ( with ketanserin and spiperone ) , 5- Q9H205 ( with MDL-72222 and ICS-205-930 ) , or Q13639 ( with ICS-205-930 ) receptors . Neither was 5-MT inhibition of P12931 affected by blocking P08908 ( with propranolol and spiperone ) , P28222 ( with propranolol ) , or P28335 ( with ketanserin ) receptors . Furthermore , 5-HT and 5-MT inhibitions of P12931 were enhanced by cilostazol [ a selective adenosine 3',5'-cyclic monophosphate ( DB02527 ) phosphodiesterase inhibitor ] and were diminished by KT-5720 ( a DB02527 -dependent protein kinase inhibitor ) but were not affected by M & B-22948 [ a selective guanosine 3',5'-cyclic monophosphate ( cGMP ) phosphodiesterase inhibitor ] or KT-5823 ( a cGMP-dependent protein kinase inhibitor ) . Biochemical studies further demonstrated that 5-HT inhibition of P12931 in porcine pial veins was accompanied by an increase in DB02527 , but not cGMP , synthesis. ( ABSTRACT TRUNCATED AT 250 WORDS ) The effects of pertussis toxin on dopamine D2 and serotonin P08908 autoreceptor-mediated inhibition of neurotransmitter synthesis : relationship to receptor reserve . Irreversible inactivation of striatal D2 dopamine ( DA ) autoreceptors with N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline ( EEDQ ) or inactivation of striatal guanine nucleotide binding proteins ( G proteins ) with pertussis toxin ( PT ) shifted the dose-response curve for N-n-propylnorapomorphine ( NPA ) -mediated inhibition of DB04699 ( Q9BVC4 ) -induced elevation of DB01235 ( DB01235 ) to the right , with a decrease in the maximum response . For the partial agonist (+)-3-(3-hydroxyphenyl)-N-n-propylpiperidine [ (+)-3-PPP ] , in contrast , there was little shift in the ED50 , after inactivation of either D2 receptors or G proteins . Completely analogous effects were found at the somatodendritic P08908 autoreceptor in the raphe nuclei , mediating inhibition of the synthesis of serotonin ( 5-HT ) ; the full agonist , 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) and the partial agonist , buspirone were utilized to inhibit the synthesis of 5-HT , as measured by changes in levels of L-5-hydroxytryptophan ( 5-HTP ) . Additionally , in both systems , combined treatment with pertussis toxin , followed by EEDQ , reduced the maximum effect , when compared to either agent alone but had little further effect on the ED50 . In systems exhibiting a large receptor reserve for agonists , such as those described above , the same pattern of response seen after inactivation of receptors or G proteins may reflect the operation of a common mechanism underlying the phenomenon of receptor reserve . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . Explorative immunohistochemical study to evaluate the addition of a topical corticosteroid in the early phase of alefacept treatment for psoriasis . The aim of this study was to explore the additional effect of betamethasone dipropionate cream in the early phase of an intramuscular ( IM ) alefacept course , on plaque severity and on modulating T-cell subsets , cells expressing NK-receptors , epidermal proliferation and keratinocyte differentiation in lesional psoriatic skin . Therefore , sixteen patients with moderate-to-severe chronic plaque psoriasis received 15 mg alefacept IM for 12 weeks , followed by a 12-week follow-up period . The first 4 weeks , patients were randomized 1:1 to either betamethasone dipropionate , or the vehicle cream , once daily . Plaque severity ( SUM ) was assessed and serial biopsies were immunohistochemically stained for T-cell subsets ( CD3 , P01730 , CD8 , CD45RO , CD45RA , P06729 , CD25 , Q9Y5U5 ) , cells expressing NK-receptors ( Q13241 and CD161 ) , epidermal proliferation ( Ki67 ) and differentiation ( P13645 ) , which were quantified using manual and digital image analysis . DB00092 monotherapy resulted in statistically significant improvement in plaque severity . Subsequently , immunohistochemical assessments on T-cell subsets , epidermal proliferation ( Ki67 ) and keratinization ( P13645 ) revealed marked time-related improvements with respect to the mentioned parameters , without significant differences between both treatment regimens . DB00092 monotherapy induces improvement of plaque severity , which is accompanied by a reduction in activated ( P06729 + , CD25+ , CD45RO+ ) dermal P01730 + and activated epidermal CD8+ T cells , epidermal proliferation and differentiation . Once daily treatment with betamethasone dipropionate cream during the first 4 weeks of an intramuscular alefacept course did not provide substantial additional clinical and immunohistochemical improvement . DB01079 and other serotonergic agents : what is the evidence ? Through effects on gastrointestinal motor and secretory function as well as visceral sensation , serotonin ( 5-HT ) plays a key role in the pathogenesis of irritable bowel syndrome ( IBS ) . In particular , 5- Q9H205 and Q13639 receptors appear to be very important in IBS . This article critically appraises the evidence supporting the use of the 5- Q9H205 receptor antagonist alosetron in the treatment of women with diarrhea-predominant IBS . The safety profile and restricted-use program for alosetron is also reviewed . This discussion is followed by a comprehensive review of the efficacy and safety data in support of tegaserod for women with constipation-predominant IBS . The effect of 1,25-dihydroxyvitamin D3 on lymphoma cell lines and expression of vitamin D receptor in lymphoma . 1,25(OH)2D3 promotes differentiation and has an antiproliferative effect in a variety of cell lines derived from the immunohaematopoetic system . alpha-Calcidol which is metabolised to 1,25(OH)2D3 has been shown to produce tumour regression in follicular low grade non-Hodgkin 's lymphoma ( Q9NZ71 ) and the dose limiting toxicity is hypercalcaemia . The cellular action of 1,25(OH)2D3 is mediated by binding to an intracellular protein , the vitamin D receptor ( P11473 ) . We have evaluated the activity of 1,25(OH)2D3 and its non-calcaemogenic analogue MC903 in the SU-DHL4 and SU-DUL5 B cell lines which carry the 14;18 translocation characteristic of follicular Q9NZ71 , and also the expression of the P11473 in a range of B cell NHLs . Both agents induced differentiation and had an antiproliferative effect on the SU-DHL4 and SU-DUL5 cell lines . However this occurred at a relatively high concentration ( 10(-7) M ) which exceeds the physiological concentration of 1,25(OH)2D3 by approximately 10(3)-10(4)-fold . Expression of the P11473 was low in each cell line and in the low grade lymphoma tumour samples . To account for the observed clinical response to 1 alpha OHD3 ( alpha-calcidol ) in follicular Q9NZ71 a network is suggested whereby 1,25(OH)2D3 modulates the activity of P01730 +T cells which have previously been shown to promote follicle centre cell proliferation . DB00169 analogues may enable serum levels to be achieved which produce a direct action on follicular lymphoma cells without disturbing calcium metabolism . Characterization of a novel Q13639 receptor antagonist of the azabicycloalkyl benzimidazolone class : DAU 6285 . Three chemical classes of serotonin Q13639 receptor agonists have been identified so far : 5-substituted indoles ( e.g. 5-HT ) , benzamides ( e.g. renzapride ) and benzimidazolones ( e.g. BIMU 8 ) . In a search for Q13639 receptor antagonists , we have discovered that the benzimidazolone derivative DAU 6285 ( for structure see text ) , is 3-5 times more potent than tropisetron in blocking 5-HT , renzapride and BIMU 8 induced stimulation of adenylate cyclase activity in mouse embryo colliculi neurons . Schild plot analysis yielded Ki values of 220 , 181 and 255 nmol/l , respectively . In addition , DAU 6285 showed poor activity as a 5- Q9H205 receptor ligand with respect to tropisetron , as demonstrated by in vitro binding studies ( Ki , 322 vs 2.8 nmol/l ) and by its antagonistic activity in the Bezold-Jarisch reflex test ( ID50 , 231 vs 0.5 micrograms/kg , i.v. ) . No significant binding ( Ki greater than 10 mumol/l ) of DAU 6285 to serotonergic P08908 , P28222 , P28335 , P28221 , and 5-HT2 receptors as well as to adrenergic alpha 1 , alpha 2 , dopaminergic D1 , D2 or muscarinic M1-M3 receptor subtypes was found . The data indicate that DAU 6285 has a somewhat higher affinity than tropisetron for Q13639 receptors , a property confirmed in functional tests , and much lower affinity than tropisetron for 5- Q9H205 receptors . The compound represents a new interesting tool for investigating the pharmacological and physiological properties of Q13639 receptors . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . Leishmania major protein disulfide isomerase as a drug target : enzymatic and functional characterization . Leishmaniasis is a major health problem worldwide and tools available for their control are limited . Effective vaccines are still lacking , drugs are toxic and expensive , and parasites develop resistance to chemotherapy . In this context , new antimicrobials are urgently needed to control the disease in both human and animal . Here , we report the enzymatic and functional characterization of a Leishmania virulence factor , Leishmania major Protein disulfide isomerase ( LmPDI ) that could constitute a potential drug target . LmPDI possesses domain structure organization similar to other P07237 family members ( a , a ' , b , b ' and c domains ) , and it displays the three enzymatic and functional activities specific of P07237 family members : isomerase , reductase and chaperone . These results suggest that LmPDI plays a key role in assisting Leishmania protein folding via its capacity to catalyze formation , breakage , and rearrangement of disulfide bonds in nascent polypeptides . Moreover , DB00626 , a reductase activity inhibitor , and DB03615 , a chaperone activity inhibitor , were tested in LmPDI enzymatic assays and versus Leishmania promastigote in vitro cultures and Leishmania amastigote multiplication inside infected THP-1-derived macrophages . DB00626 inhibited both isomerase and reductase activities , while DB03615 had no effect on the chaperone activity . Interestingly , DB00626 blocked in vitro promastigote growth as well as amastigote multiplication inside macrophages with EC(50) values of 39 μM . These results suggest that LmPDI may constitute an interesting target for the development of new anti-Leishmania drugs . The role of endothelium-derived hyperpolarizing factor and cyclooxygenase pathways in the inhibitory serotonergic response to the pressor effect elicited by sympathetic stimulation in chronic sarpogrelate treated rats . We have demonstrated that the antagonism of 5-HT2 receptors produces an enhancement of serotonergic sympathoinhibitory effect by P28221 and P34969 activation . The aim of this work was to determine mechanisms involved in the 5-hydroxytriptaminergic inhibitory action on the pressor responses elicited by sympathostimulation in pithed rats treated with a 5-HT2 receptor blocker . The blockade of 5-HT2 receptors was induced by orally sarpogrelate treatment ( 30 mg/kg/day ) . Two weeks later , animals were anaesthetized and pithed . A bolus injection of 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one ( ODQ ) ( 10 µg/kg ) , a guanylyl cyclase inhibitor , or indomethacin ( 2mg/kg ) , a non-selective P36551 inhibitor , prior to the infusion of ( 2S ) (+)-5-(1,3,5-trimethylpyrazol-4-yl)-2-(dimethylamino)tetralin , AS-19 ( 5 µg/kg/min ) were not able to abolish its inhibitory action . However , i.v. administration of glibenclamide ( 20mg/kg ) , a blocker of DB00171 -sensitive K(+) channels , completely reversed AS-19 sympathoinhibitory action . The inhibitory effect of 2-[5-[3-(4-methylsulfonylamino)benzyl-1,2,4-oxadiazol-5-yl]-1H-indol-3-yl]ethanamine , L-694,247 ( 5 µg/kg/min ) was abolished by indomethacin , whereas pretreatment with ODQ had no effect . DB04743 ( 3mg/kg ) , a P35354 inhibitor , completely reversed the inhibitory action of L-694,247 , whereas 1- [ [ 4,5-bis (4-methoxyphenyl)-2-thiazolyl ] carbonyl ] -4-methylpiperazine hydrochloride ( FR122047 ) ( 3mg/kg ) , a P23219 inhibitor , partially blocked this action . The sympathoinhibition by 5-HT ( 20 µg/kg/min ) could not be elicited after i.v. treatment with indomethacin plus glibenclamide . In conclusion , these results suggest that in chronic sarpogrelate-treated rats , the inhibitory serotonergic effect of the pressor responses induced by electrical stimulation of the sympathetic outflow via P34969 and P28221 receptor activation is mediated by KATP channel-mediated smooth muscle hyperpolarization and the P36551 pathway , respectively . P35372 -dependent and independent components in effects of tramadol . DB00193 is thought to induce analgesia via both opioid and non-opioid pathways , although the precise mechanisms remain to be elucidated . In this study , we investigated the roles of the mu-opioid receptor ( MOP ) in analgesic and rewarding effects of tramadol by using MOP knockout ( KO ) mice . DB00193 -induced antinociception , assessed by hot-plate and tail-flick tests , was significantly reduced in heterozygous and homozygous MOP-KO mice when compared with that in wild-type mice . Interestingly , however , tramadol retained its ability to induce significant antinociception in homozygous MOP-KO mice . The tramadol-induced antinociception remaining in homozygous MOP-KO mice was not significantly affected by methysergide , a serotonin receptor antagonist , but was partially blocked by yohimbine , an adrenaline alpha2 receptor antagonist , and both naloxone , a non-selective opioid receptor antagonist , and yohimbine . In addition , antinociceptive effects of an active tramadol metabolite M1 were abolished or remarkably reduced in MOP-KO mice . On the other hand , neither wild-type nor homozygous MOP-KO mice showed significant place preference for tramadol in a conditioned place preference test , although there were slight tendencies toward preference in wild-type mice and avoidance in homozygous MOP-KO mice . These results strongly support the idea suggested in the previous pharmacological studies that MOP and the adrenaline alpha2 receptor mediate most of the analgesic properties of tramadol . DB00092 ( anti- P06729 ) causes a selective reduction in circulating effector memory T cells ( Tem ) and relative preservation of central memory T cells ( Tcm ) in psoriasis . BACKGROUND : DB00092 ( anti- P06729 ) biological therapy selectively targets effector memory T cells ( Tem ) in psoriasis vulgaris , a model Type 1 autoimmune disease . METHODS : Circulating leukocytes were phenotyped in patients receiving alefacept for moderate to severe psoriasis . RESULTS : In all patients , this treatment caused a preferential decrease in effector memory T cells ( P32248 - CD45RA- ) ( mean 63 % reduction ) for both P01730 + and CD8+ Tem , while central memory T cells ( Tcm ) ( P32248 +CD45RA- ) were less affected , and naïve T cells ( P32248 +CD45RA+ ) were relatively spared . Circulating CD8+ effector T cells and Type 1 T cells ( P01579 -producing ) were also significantly reduced . CONCLUSION : DB00092 causes a selective reduction in circulating effector memory T cells ( Tem ) and relative preservation of central memory T cells ( Tcm ) in psoriasis . Production of an antiserum specific to the ADP-ribosylated form of elongation factor 2 from archaebacteria and eukaryotes . An antiserum to ADP-ribosylated elongation factor 2 ( DB02059 - P13639 ) from S. acidocaldarius was raised in rabbits using stained , homogenized , DB02059 - P13639 -containing slices from SDS-gels as a source of antigen . P13639 ( P13639 ) from S. acidocaldarius was cloned in E. coli and the expressed gene product was used in order to adsorb all anti- P13639 antibodies which do not contain the ADP-ribosyl group within their epitopes , as E. coli is unable to synthesize the ADP-ribosyl acceptor diphthamide . The remaining antibodies were specific to ADP-ribosylated P13639 from Thermoplasma acidophilum , S. acidocaldarius and Desulfurococcus mucosus . ADP-ribosylated P13639 from eukaryotic sources also reacted with the adsorbed antiserum as shown for P13639 isolated from the killi-fish Cynolebias whitei , the mouse species BALB/c and Han/Wistar rats . The adsorbed antiserum did not cross-react with ADP-ribosylated actin or rho protein or with DB03147 -containing D-amino acid oxidase . Organization and nucleotide sequence of a transcriptional unit of Methanococcus vannielii comprising genes for protein synthesis elongation factors and ribosomal proteins . By a chromosome walking strategy the DNA region from Methanococcus vannielii flanking the genes for protein synthesis elongation factor ( EF ) 1 alpha and P13639 was cloned and sequenced . A gene organization of 5' - beta ' - open reading frame ( ORF ) 1 - ORF2 - P28222 - S7 - P13639 - EF-1 alpha - S10 - ORF3 - ORF4 - 3' was found where beta ' , P28222 , S7 , S10 , P13639 , and EF-1 alpha represent gene products with sequences similar to the beta ' subunit of RNA polymerase , ribosomal proteins P28222 , S7 , and S10 , and EF-G and EF-Tu from Escherichia coli , respectively . ORF1-4 represent gene products with no known eubacterial counterparts . Northern blot analysis of transcripts and nuclease S1 mapping showed that transcription initiates between beta ' and ORF1 and terminates at the 3' side of the S10 gene and that the genes from ORF1 to S10 are cotranscribed . Apart from the presence of two additional ORFs , ORF1 and ORF2 , and of the gene for S10 , this organization is identical to that of the eubacterial " streptomycin operon. " ORF1 displays sequence similarity to rat liver ribosomal protein L30 and may represent one of the " additional " ribosomal proteins of Methanococcus . The sequenced part of the beta ' gene and the P13639 and EF-1 alpha gene products from Methanococcus are more similar to their eukaryotic than to their eubacterial counterparts . It appears , therefore , that the genetic organization of the translational components resembles the situation in eubacteria , whereas their primary structures are more eukaryotic in nature .

MH_train_1376

MH_train_1376

MH_train_1376

interacts_with DB01406?

multiple_choice

Therapeutic targeting of the PDGF and TGF-beta-signaling pathways in hepatic stellate cells by PTK787/ZK22258 . Stimulation of hepatic stellate cells ( HSCs ) by platelet-derived growth factor ( PDGF ) and transforming growth factor-beta1 ( TGF-beta1 ) is an essential pathway of proliferation and fibrogenesis , respectively , in liver fibrosis . We provide evidence that PTK787/ZK222584 ( DB04879 ) , a potent tyrosine kinase inhibitor that blocks vascular endothelial growth factor receptor ( VEGFR ) , significantly inhibits PDGF receptor expression , as well as PDGF-simulated P19526 proliferation , migration and phosphorylation of P27361 /2 , Akt and p70S6 kinase . Interestingly , DB04879 also antagonizes the TGF-beta1-induced expression of P15692 and P17948 . Furthermore , DB04879 downregulates TGF-beta receptor expression , which is associated with reduced Akt , P29323 and p38MAPK phosphorylation . Furthermore , PDGF-induced TGF-beta1 expression is inhibited by DB04879 . These findings provide evidence that DB04879 targets multiple essential pathways of stellate cell activation that provoke proliferation and fibrogenesis . Our study underscores the potential use of DB04879 as an antifibrotic drug in chronic liver disease . DB00644 agonist inhibits human telomerase reverse transcriptase mRNA expression in endometrial cancer cells . We investigated the relationship between the antiproliferative effect of DB00644 agonist and telomerase activity using the endometrial cancer cell line O14777 -1A . The subjects were 38 endometrial cancer , and 2 atypical endometrial hyperplasia patients . P30968 expression was detected using RT-PCR . O14777 -1A cells were incubated with 10(-7)-10(-4) M DB00644 agonist ( leuprolide acetate ) , and cell proliferation was determined using MTT assay . The telomerase activity was detected by the TRAP assay and expression of human telomerase reverse transcriptase ( hTERT ) was assessed by RT-PCR . P30968 mRNA was detected at 94.7 % ( 36/38 ) in endometrial cancer and in both of the atypical endometrial hyperplasia and in O14777 -1A cells . Cell proliferation of O14777 -1A showed significant inhibition at leuprolide acetate concentrations of 10(-6) M or higher compared with untreated control culture ( p < 0.05 ) . The telomerase activity showed no marked difference compared with untreated culture . However , hTERT mRNA expression showed a decrease in the leuprolide-treated cells . It is suggested that the mechanism of the antitumor effect of DB00644 agonist involved the inhibition of hTERT mRNA expression in the endometrial cancer cells . Ghrelin promotes intestinal epithelial cell proliferation through PI3K/Akt pathway and P00533 trans-activation both converging to P29323 1/2 phosphorylation . Little is known about ghrelin 's effects on intestinal epithelial cells even though it is known to be a mitogen for a variety of other cell types . Because ghrelin is released in close proximity to the proliferative compartment of the intestinal tract , we hypothesized that ghrelin may have potent pro-proliferative effect on intestinal epithelial cells as well . To test this hypothesis , we characterized the effects of ghrelin on FHs74Int and Caco-2 intestinal epithelial cell lines in vitro . We found that ghrelin has potent dose dependent proliferative effects in both cell lines through a yet to be characterized G protein coupled growth hormone secretagogue receptor ( Q92847 ) subtype . Consistent with above findings , cell cycle flowcytometric analyses demonstrated that ghrelin shifts cells from the P55008 to S phase and thereby promotes cell cycle progression . Further characterization of subcellular events , suggested that ghrelin mediates its pro-proliferative effect through DB00131 cyclase ( AC ) -independent epidermal growth factor receptor ( P00533 ) trans-activation and PI3K-Akt phosphorylation . Both these pathways converge to stimulate MAPK , P29323 1/2 downstream . The role of ghrelin in states where intestinal mucosal injury and rapid mucosal repair occur warrants further investigation . P50579 is required for P19526 initiation and proliferation . In a chemical screening , we tested the antiangiogenic effects of fumagillin derivatives and identified fumagillin as an inhibitor of definitive hematopoiesis in zebrafish embryos . DB02640 is known to target methionine aminopeptidase II ( MetAP2 ) , an enzyme whose function in hematopoiesis is unknown . We investigated the role of MetAP2 in hematopoiesis by using zebrafish embryo and human umbilical cord blood models . Zebrafish metap2 was expressed ubiquitously during early embryogenesis and later in the somitic region , the caudal hematopoietic tissue , and pronephric duct . metap2 was inhibited by morpholino and fumagillin treatment , resulting in increased mpo expression at 18 hours postfertilization and reduced c-myb expression along the ventral wall of dorsal aorta at 36 hours postfertilization . It also disrupted intersegmental vessels in Tg(fli1:gfp) embryos without affecting development of major axial vasculatures . Inhibition of MetAP2 in CB P28906 (+) cells by fumagillin had no effect on overall clonogenic activity but significantly reduced their engraftment into immunodeficient nonobese diabetes/severe combined immunodeficiency mice . metap2 knock-down in zebrafish and inhibition by fumagillin in zebrafish and human CB P28906 (+) cells inhibited P62158 Kinase II activity and induced P29323 phosphorylation . This study demonstrated a hitherto-undescribed role of MetAP2 in definitive hematopoiesis and a possible link to noncanonical Wnt and P29323 signaling . P01308 action on H292 bronchial carcinoma cells as compared to normal bronchial epithelial cells . DB00030 may contribute to bronchial carcinoma due to P08069 activation by high local concentrations . Therefore , effects of insulin and P05019 on human bronchial carcinoma cells ( H292 ) and normal bronchial epithelium cells ( P02100 ) were studied . TGF-β was included since it also influences carcinoma progression . H292 and P02100 cells expressed both the insulin receptor and the P08069 ; in H292 cells an additional , shorter , splicing variant ( IR-A ) of the insulin receptor was present . P06213 expression was around four to five times higher in H292 than in P02100 cells at mRNA and protein levels . P01308 and TGF-β exerted contrary actions on proliferation and gene expression in H292 cells . Genes regulated by insulin , P05019 , and TGF-β were linked to inflammation , cell adhesion , muscle contraction and differentiation . P01308 and P05019 also suppressed DNA repair genes . EC(50) for insulin-induced proliferation was around 5 nM in H292 and around 30 nM P02100 cells . The EC(50) values for gene expression ranged from 9 to 90 nM in both cell types , dependent on the gene studied . In H292 cells , the proliferative response was much stronger if TGF-β was present . In P02100 cells this interaction of insulin and TGF-β was not observed , and changes in gene expression were mostly lower by at least 10-fold as compared to H292 . All in all , the insulin effects in H292 were generally much stronger than in P02100 cells and - with regard to proliferation - occurred at lower concentrations . Thus , insulin will hardly induce cancer from normal bronchial cells but may favour progression of pre-existing tumours . Analysis of breast cancer related gene expression using natural splines and the Cox proportional hazard model to identify prognostic associations . Many studies correlating gene expression data to clinical parameters assume a linear increase or decrease of the clinical parameter under investigation with the expression of a gene . We have studied genes encoding important breast cancer-related proteins using a model for survival-type data that is based on natural splines and the Cox proportional hazard model , thereby removing the linearity assumption . Expression data of 16 genes were studied in relation to metastasis-free probability in a cohort of 295 consecutive breast cancer patients treated at The Netherlands Cancer Institute . The independent predictive power for disease outcome of the 16 individual genes was tested in a multivariable model with known clinical and pathological risk factors . There is a linear relationship between increasing expression and a higher or lower hazard for distant metastasis for P03372 , Q15303 , P15692 , O96020 , Q15910 , and Q96NZ9 ; for P04626 , P21860 , P24385 , P24864 , O75530 , P61073 , P32248 , P48061 , and P05121 there is no clear increase or decrease ; and for P00533 there seems to be a non-linear relation . Multivariable analysis showed that the 70-gene prognosis profile outperforms all the other variables in the model ( hazard-rate 5.4 , 95 % CI 2.5-11.7 ; P = 0.000018 ) . P00533 -expression seems to have a non-linear relation with disease outcome , indicating that lower but also higher expression of P00533 are associated with worse outcome compared to intermediate expression levels ; the other genes show no or a linear relation . Expression of the human concentrative nucleotide transporter 1 ( O00337 ) gene correlates with clinical response in patients affected by Waldenström 's Macroglobulinemia ( WM ) and small lymphocytic lymphoma ( SLL ) undergoing a combination treatment with 2-chloro-2'-deoxyadenosine ( DB00242 ) and DB00073 . PURPOSE : Resistance to nucleoside analogues agents is likely to be multifactorial and could involve a number of mechanisms affecting drug penetration , metabolism and targeting . In vitro studies of resistant human cell lines have confirmed that human concentrative nucleoside transporter 1 ( O00337 ) -deficient cells display resistance . EXPERIMENTAL DESIGN : We applied real-time PCR method to assess the mRNA expression of equilibrative and concentrative nucleoside transporter ( hENT1 , O00337 ) , deoxycytidine and deoxyguanosine kinase ( P27707 , Q16854 ) , 5'-nucleotidase ( 5'-NT ) , ribonucleotide reductase catalytic and regulatory ( P23921 , P31350 ) subunits in bone marrow cells from 32 patients with Waldenström 's Macroglobulinemia ( WM ) and small lymphocytic lymphoma ( SLL ) who received 2CdA-based chemotherapy . Responses to chemotherapy , were then correlated to the expression of these markers . RESULTS : All 32 patients enrolled expressed lower levels of O00337 as compared to healthy donors . In univariate analysis , lower expression level of O00337 ( p=0.0021 ) and P31350 ( p=0.02 ) correlated with response to chemotherapy . In particular , patients with low levels of O00337 achieved inferior clinical response . No significant correlation between these genes expression and age , stage of disease was found . This study suggests that nucleotidase expression levels can be used to identify subgroups of WM and SLL patients who will likely respond differently to a 2CdA-based therapy . Adenoid cystic carcinomas of the breast have low Topo IIα expression but frequently overexpress P00533 protein without P00533 gene amplification . Adenoid cystic carcinoma of the breast is a rare subtype of breast cancer with basal-like features . Published studies on breast adenoid cystic carcinoma are limited , resulting in relatively scarce information on the value of predictive tumor markers . We studied 20 primary cases of adenoid cystic carcinoma of the breast for expression of estrogen receptor , progesterone receptor , androgen receptor , epidermal growth factor receptor , HER-2/neu , and topoisomerase IIα using immunohistochemistry and fluorescent in situ hybridization methods . Estrogen and progesterone receptor expression were detected in 1 case each . All tumors were uniformly negative for Her-2/neu expression . P10275 and topoisomerase IIα expression were weakly positive in three cases and 7 cases , respectively . P00533 overexpression was detected in 13 cases ( 65 % of all cases ) . Amplification of P11388 or HER-2/neu gene was not detected in any of the cases . Our study shows that the majority of adenoid cystic carcinomas of the breast do not overexpress Her-2/neu , topoisomerase IIα , or estrogen receptor , and thus , they are unlikely to respond to therapies targeting these proteins . However , these tumors frequently over-express epidermal growth factor receptor , indicating a potential benefit from anti-epidermal growth factor receptor therapy for patients with advanced adenoid cystic carcinomas of the breast . [ 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 . Q92847 agonist ( DB05657 ) accelerates gastric emptying in adults with diabetes and symptomatic gastroparesis . BACKGROUND : DB05657 is a synthetic , selective ghrelin agonist in development for gastroparesis . AIM : To assess safety and effects of DB05657 in diabetes patients with symptomatic gastroparesis . METHODS : Adults with type 1 or type 2 diabetes mellitus received placebo and DB05657 ( 80 , 160 , 320 or 600 microg/kg ) infusions in a cross-over manner following a radiolabelled meal . Blood glucose levels were stabilized using a hyperinsulinemic-euglycemic clamp . Primary endpoints were gastric half emptying and latency times . Secondary measures included assessment of gastroparesis symptoms and endocrine responses . RESULTS : Ten patients with type 1 ( n = 7 ) or 2 ( n = 3 ) diabetes , moderate-to-severe gastroparesis symptoms and > or =29 % retention 4 h after a radiolabelled solid meal were enrolled . DB05657 produced significant reductions in solid meal half-emptying ( 20 % , P = 0.043 ) and latency ( 34 % , P = 0.037 ) times vs. placebo . Reductions in overall postmeal symptom intensity ( 24 % ) and postprandial fullness ( 37 % ) following DB05657 infusion were not statistically significant . Most adverse events were mild and self-limiting and there were no identifiable differences in numbers or types of adverse events between DB05657 and placebo . CONCLUSIONS : This proof-of-concept study demonstrates that the ghrelin agonist DB05657 is well-tolerated in diabetes patients with moderate-to-severe chronic gastroparesis and shows statistically significant improvements in gastric emptying . 3-deazaadenosine inhibits leukocyte adhesion and P05362 biosynthesis in tumor necrosis factor-stimulated human endothelial cells . Previous reports demonstrate that cultured human umbilical vein endothelial cells ( O14777 ) treated with P01375 and other inflammatory mediators show an increased capacity to adhere human neutrophils . This increase is associated with the up-regulation of intercellular adhesion molecule 1 ( P05362 ) and other adhesion molecules on the O14777 surface . We have found that 200 microM 3-deazaadenosine ( c3Ado ) prevented this P01375 -induced increase in O14777 adhesiveness . This effect resulted from interactions of c3Ado with O14777 and not with polymorphonuclear neutrophils . Transport of c3Ado into the O14777 was required for its activity , as evidenced by antagonism with the nucleoside transport inhibitor , nitrobenzylthioinosine . Treatment of O14777 with c3Ado led to the intracellular buildup of S-adenosylhomocysteine and to the metabolic formation of S-3-deazaadenosylhomocysteine and 3-deazaadenosine 5'-triphosphate , events that appeared not to contribute to c3Ado activity . Exogenous L-homocysteine potentiated c3Ado activity , and this potentiation was prevented by the S-adenosylhomocysteine hydrolase inhibitor , periodate-oxidized adenosine . By using the mAb P23921 /1 , we have determined that c3Ado also inhibited the P01375 -induced expression of P05362 on the surface of the O14777 , as well as cytosol-associated P05362 . Northern blot and in vitro translation analyses of poly(A+) RNA from c3Ado-treated O14777 revealed that this nucleoside analog selectively decreased steady-state levels of P05362 mRNA . The capacity of c3Ado to selectively inhibit O14777 adhesiveness , P05362 production , and steady-state levels of P05362 mRNA may contribute to the drug 's activity as an anti-inflammatory agent . Functional expression of rat pituitary gonadotrophin-releasing hormone receptors in Xenopus oocytes . Expression of receptors for the hypothalamic regulatory peptide , gonadotrophin-releasing hormone ( DB00644 ) , was investigated by intracellular recording from Xenopus oocytes injected with poly(A)+ mRNA isolated from rat anterior pituitary glands . Membrane depolarizations were induced in oocytes in a dose-dependent fashion following the application of DB00644 ( 10nM - 1 microM ) or a DB00644 superactive agonist , buserelin ( 1nM - 1 microM ) . The response was reversibly blocked by the addition of a DB00644 antagonist ( 1 microM ) . TRH ( 10nM - 1 microM ) had no effect on most of these oocytes . In contrast , some other oocytes which showed no responses to DB00644 or to the DB00644 agonist , displayed depolarizing responses to TRH ( 10nM - 1 microM ) . A relatively small number of oocytes responded to both ligands . Control oocytes did not respond to the DB00644 analogues or to TRH . This successful expression of the P30968 could provide a new approach to the study of the receptor , and serve as a means for the isolation and cloning of the encoding genes . Q08462 selectively couples to E prostanoid type 2 receptors , whereas adenylyl cyclase 3 is not receptor-regulated in airway smooth muscle . Adenylyl cyclases ( ACs ) are important regulators of airway smooth muscle function , because β-adrenergic receptor ( βAR ) agonists stimulate AC activity and DB02527 production . We have previously shown in a number of cell types that AC6 selectively couples to βAR and these proteins are coexpressed in lipid rafts . We overexpressed AC2 , O60266 , and AC6 in mouse bronchial smooth muscle cells ( mBSMCs ) and human embryonic kidney ( P29320 ) -293 cells by using recombinant adenoviruses and assessed their localization and regulation by various G protein-coupled receptors ( GPCRs ) . O60266 and AC6 were expressed primarily in caveolin-rich fractions , whereas AC2 expression was excluded from these domains . AC6 expression enhanced DB02527 production in response to isoproterenol but did not increase responses to butaprost , reflecting the colocalization of AC6 with β(2)AR but not E prostanoid type 2 receptor ( EP(2)R ) in lipid raft fractions . AC2 expression enhanced butaprost-stimulated DB02527 production but had no effect on the β(2)AR-mediated response . O60266 did not couple to any GPCR tested . DB02587 -induced arborization of mBSMCs was assessed as a functional readout of DB02527 signaling . Arborization was enhanced by overexpression of AC6 and O60266 , but AC2 had no effect . GPCR-stimulated arborization mirrored the selective coupling observed for DB02527 production . With the addition of the phosphodiesterase 4 ( DB05876 ) inhibitor rolipram AC2 accelerated forskolin-stimulated arborization . Thus , AC2 selectively couples to EP(2)R , but signals from this complex are limited by DB05876 activity . O60266 does not seem to couple to GPCR in either mBSMCs or P29320 -293 cells , so it probably exists in a distinct signaling domain in these cells . Mite antigens enhance P05362 and induce P19320 expression on human umbilical vein endothelium . Although sublingual allergen-specific immunotherapy has been proved to be effective in the treatment of allergic diseases , controversy surrounds the means by which such a local therapy can induce systemic immunological changes . Adhesion molecules are critical in the regulation of leukocyte traffic . It has been hypothesized that allergenic extract , administered locally , may induce an up-regulation of the mucosal vessel vascular adhesion molecules ( CAMs ) resulting in local recruitment of circulating inflammatory cells . In the present study we investigated whether the mite antigens , Der p1 and Der p2 , can modulate P62158 expression of human endothelial cells ( O14777 ) . To do this , slices of whole human umbilical cord vein underwent short-term ( 8 hours ) cultures in the presence or absence of mite antigen ( baseline , unstimulated controls ) . Cryostatic sections of the specimens were then evaluated immunohistochemically for expression of intercellular adhesion molecule ( P05362 ) and vascular cell adhesion molecule ( P19320 ) molecules . The results revealed that while Der p1 is capable of significantly up-regulating P05362 and P19320 on O14777 , Der p2 antigen moderately up-regulates P05362 expression but is ineffective in modulating P19320 . Although preliminary , these results clearly support the hypothesis that at least some of the effects of sublingual immunotherapy may derive from inflammatory cell recruitment at the site of allergen release . P10275 repression of gonadotropin-releasing hormone gene transcription via enhancer 1 . DB00644 ( DB00644 ) plays a major role in the hypothalamic-pituitary-gonadal ( HPG ) axis , and synthesis and secretion of DB00644 are regulated by gonadal steroid hormones . Disruptions in androgen levels are involved in a number of reproductive defects , including hypogonadotropic hypogonadism and polycystic ovarian syndrome . Androgens down-regulate DB00644 mRNA synthesis in vivo and in vitro via an androgen receptor ( AR ) -dependent mechanism . DB02998 ( R1881 ) , a synthetic AR agonist , represses DB00644 expression through multiple sites in the proximal promoter . In this study , we show AR also represses DB00644 transcription via the major enhancer ( DB00644 -E1 ) . A multimer of the -1800/-1766 region was repressed by R1881 treatment . Mutation of two bases , -1792 and -1791 , resulted in decreased basal activity and a loss of AR-mediated repression . AR bound to the -1796/-1791 sequence in electrophoretic mobility shift assays , indicating a direct interaction with DNA or other transcription factors in this region . We conclude that AR repression of DB00644 -E1 acts via multiple AR-responsive regions , including the site at -1792/-1791 . The infection of human primary cells and cell lines by human cytomegalovirus : new tropism and new reservoirs for HCMV . Human cytomegalovirus ( HCMV ) infection is asymptomatic in common persons and could reactive in immunosuppression groups . HCMV was considered as endothelial cells ( EC ) tropism and leukocyte tropism . We hypothesized that HCMV will infect other cell types from human which have not been reported yet . The HCMV released from human MRC-5 was inoculated into eight human primary cells and cell lines , including human dermal fibroblasts ( HDF ) , human embryo-chondrocytes ( O14777 ) , human embryo-myoblasts ( HEM ) , and human embryo-kidney endothelial cell ( P29320 -EC ) , human marrow stromal cell ( HMSC ) . The cell lines were ECV304 , Chung liver cell and L02 . Several detection methods specific for HCMV , in which PCR for HCMV DNA sequences , immunofluorescence for pp65 antigen , Western-blot for gB protein , as well as cytopathic effect observation were conducted at different time post-infection . The results indicated that four cells in our experiment ( HDF , HEM , O14777 and HMSC ) were HCMV-positive . The occurring time of cytopathic effect was different in these four cells . Our experiment found the new tropism and new reservoirs for HCMV . P05362 -independent lymphocyte transmigration across high endothelium : differential up-regulation by interferon gamma , tumor necrosis factor-alpha and interleukin 1 beta . The adhesion of lymphocytes to cytokine-treated high endothelium was studied using cultured high endothelial cells ( O14777 ) . Pretreatment of the O14777 layer with a variety of cytokines caused up-regulation of lymphocyte adhesion with the effects ordered interferon gamma ( P01579 ) greater than tumor necrosis factor-alpha ( P01375 ) greater than or equal to interleukin 1 beta ( IL 1 beta ) . Increased lymphocyte adhesion was found to be independent of P05362 as expression by O14777 was not increased by cytokines and antibodies against P05362 did not block adhesion . The peptide CS1 and anti-beta 1 integrin subunit antibodies , however , caused partial inhibition of lymphocyte adhesion thus indicating a role for fibronectin on O14777 and alpha 4 beta 1 on lymphocytes . Study of the kinetics of lymphocyte adhesion showed that the effects of P01579 and P01375 were persistent and remained detectable 2.5 h after removal of the cytokines whereas the effects of IL 1 beta were transient and were not sustained beyond 1 h . All of the cytokines used caused transient increases in the number of surface-bound lymphocytes with P01579 greater than P01375 greater than or equal to IL 1 beta , however , the most dramatic effect was on the transmigration of lymphocytes across the O14777 . Both P01579 and P01375 caused sustained increased transmigration with P01579 having the greater effect . IL 1 beta had little effect on transmigration . This model demonstrates that the binding and transmigration of lymphocytes across O14777 can be differentially regulated by the actions of individual cytokines . These results support the concept that locally produced cytokines regulate O14777 function within the lymph node . Cytoplasmic and nuclear estrogen binding capacity in the rat uterus during treatment with danazol and testosterone . DB01406 , testosterone and dihydrotestosterone ( DB02901 ) were tested as competitors for estrogen receptors on immature rat uterus cytosol . No competitive binding could be demonstrated for any of these steroids . After that , prepubertal Wistar rats were exposed to danazol , testosterone or propylene glycol ( control ) for 3 days or 17 days . After the appropriate exposure to medication , the animals were killed . Both danazol and testosterone appeared to be uterotropic after 3 days of treatment , although the increase in the uterine weight was significant only in the danazol-treated group ( p less than 0.05 ) . This effect was lost after 17 days of treatment . P03372 binding assays were done on the cytosolic and nuclear fractions of the homogenized uterine tissue of each group . The estrogen binding capacity of cytosols was increased in both the danazol ( p less than 0.05 ) and the testosterone ( p less than 0.01 ) groups after 3 days of treatment . A parallel increase was found in the nuclear fraction of both groups . After 17 days of treatment , the comparison between the 3 groups showed no differences in the cytosolic or nuclear estrogen binding capacity . The information provided by this study suggests that some effects of danazol may be due to an androgenic action and that may be associated to increases in the free fraction of testosterone . Regulation of PTP1D mRNA by peptide growth factors in the human endometrial cell line O14777 -1-A . OBJECTIVE : To assess , in the human endometrial cell line O14777 -1-A , the presence of protein tyrosine phosphatase 1D ( PTP1D ) and the possible regulation of its mRNA expression by mitogens such as forskolin ( an agent that increases intracellular cyclic adenosine monophosphate [ DB02527 ] levels ) , epidermal growth factor ( P01133 ) , and insulin-like growth factor-I ( P05019 ) . METHODS : Cells were grown to confluence and maintained in serum-free media for 24 hours before treatment . Cells were exposed to forskolin , P01133 , and P05019 for increasing time periods ( 0 , 1 , 3 , 6 , and 24 hours ) , and PTP1D mRNA expression was determined by Northern blot analysis . In addition , cells were incubated with increasing doses of forskolin ( final concentrations : 1 , 5 , 10 , 20 , and 30 mumol/L ) for 6 hours . RESULTS : When treated with the various mitogens , cells increased their stimulation of PTP1D mRNA expression in a time- and dose-dependent fashion . Specifically , forskolin , P01133 , and P05019 induced maximal mRNA expression at 6 , 3 , and 6 hours , respectively . Expression induced by forskolin , P01133 , and P05019 was five , three , and six times control levels , respectively . At a dose of 10 mumol/L , forskolin induced PTP1D mRNA expression almost two times higher than control values . CONCLUSION : These data suggest that in human endometrial carcinomas , DB02527 , P01133 , and P05019 may regulate the expression of PTP1D mRNA , which may , in turn , play a role in uncontrolled cell proliferation and neoplastic transformation . DB08904 : a new biologic targeting rheumatoid arthritis . The past decade has been an exciting period for clinical research and patient care in rheumatoid arthritis . This is mostly due to targeted biologic agents that have changed the outcome of this disease . DB08904 ( Cimzia(®) , UCB Inc. , GA , USA ) , which targets P01375 -α with a different mechanism of action than widely used biologics , was initially investigated for Crohn 's disease but has now been shown to be effective for rheumatoid arthritis . There have been three significant clinical trials demonstrating the efficacy of certolizumab pegol in active rheumatoid arthritis ; two with combination methotrexate and one with monotherapy . This article will summarize the data from those trials and compare some of the characteristics of certolizumab pegol to conventional disease-modifying antirheumatic drugs and other biologic agents . Treatment recommendations are beyond the scope of this review ; however , with many options available , there will be annotations on current trends in the care of this chronic disease .

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interacts_with DB00266?

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Xanthan/chitosan gold chip for metal enhanced protein biomarker detection . Protein microarrays for disease diagnostics are required to accurately quantify analytes in the low pg/mL range . This task is hampered by weak signal strengths and too low detector sensitivity . Herein we present reflective gold chips coated with polyelectrolyte multilayers ( PEMs ) for signal enhancement in immunoassays for melanoma-relevant biomarkers . Among tested (semi)natural polysaccharides ( xanthan , chitosan , carboxymethylcellulose , hyaluronic acid ) PEMs composed of xanthan and chitosan performed best in terms of detection of low analyte concentrations ( ED10 ) , spot morphology , fluorescence background and variability ( < 10 % ) . Fluorescence signals on gold slides with a 75 nm coating of seven crosslinked polyelectrolyte double layers were up to 50 times higher than on bare glass slides . In comparison to commercial substrates the signal to noise ratio is enhanced by up to factor 11 . Furthermore sandwich assays for interleukins 6 , 8 , 10 , tumour necrosis factor alpha ( TNFα ) , vascular endothelial growth factor A ( P15692 ) and P04271 show working ranges which cover significantly lower concentrations ( up to 38-fold ) . Not limited to above assays the presented substrates , which combine a biocompatible interface with metal-based signal amplification , are a valuable tool in a variety of biosensor applications . Proteomic discovery of genistein action in the rat mammary gland . DB01645 , the primary isoflavone component of soy , consumed in diet during the prepubertal period suppresses chemically induced mammary cancer in rats . The current study used two-dimensional gel electrophoresis ( 2-DE ) /MS-based proteomic technology to identify proteins responsible for genistein breast cancer protection In Vivo . Female offspring were exposed via lactating dams treated with 250 mg genistein/kg AIN-76A diet from days 1 to 21 postpartum ( prepubertal period ) . Mammary glands were collected at 21 and 50 day of age and subjected to 2-DE/MS and immuno-blot analyses . Twenty-three proteins were determined to be differentially regulated ( p < 0.05 ) and identified using 2-DE , followed by MALDI-TOF/TOF or LC- P19957 -MS/MS . Five of these proteins were validated by immuno-blots . P07355 was significantly increased at 21 days yet found to be decreased at 50 days . Fetuin B was found to be unchanged at day 21 but increased at day 50 . P00558 ( P00558 ) was unchanged at day 21 but decreased at day 50 . P06396 was increased at day 21 but not at day 50 . P30101 ( P30101 ) was decreased at day 21 and unchanged at day 50 . Also , we found that vascular endothelial growth factor receptor 2 ( P15692 -R2 ) and epidermal growth factor receptor ( P01133 -R ) were decreased in mammary glands of 50-day-old rats treated prepubertally with genistein . This study demonstrates the usefulness of proteomics for the discovery of key proteins involved in signaling pathways to understand genistein mechanisms of action in breast cancer prevention . Altered growth factor expression in the aging penis : the Brown-Norway rat model . The objective of the present study was to evaluate age-related changes in the protein and gene expression of modulators of erectile function ( nitric oxide [ NO ] and endothelin-1 [ ET-1 ] ) and growth factors such as transforming growth factor ( TGF-beta1 ) and vascular endothelial growth factor ( P15692 ) in the penile tissue of Brown-Norway ( BN ) rats . Young and old BN male rats were euthanized , and the penile tissue was processed for immunohistochemical and molecular analyses . Total RNA was extracted , and an Access reverse transcription-polymerase chain reaction ( RT-PCR ) system was used for messenger RNA ( mRNA ) expression analysis . Immunohistochemical studies showed a decreased expression of endothelial nitric oxide synthase ( P29474 ) protein and an increased staining for ET-1 . Quantitative analysis of PCR products revealed decreased levels of P15692 mRNA expression in the old population of rats . The most significant decrease was detected between bands corresponding to splice forms 164 ( 21 % ) and 120 ( 18 % ) . The observed alterations in the gene expression of growth factors such as P15692 may contribute to the abnormal age-related morphological and physiological alterations in the erectile tissue . [ Pharmacological and clinical profile of mitiglinide calcium hydrate ( Glufast ) , a new insulinotropic agent with rapid onset ] . DB01252 calcium hydrate ( mitiglinide , Glufast ) is a new insulinotropic agent of the glinide class with rapid onset . DB01252 is thought to stimulate insulin secretion by closing the DB00171 -sensitive K(+) ( K( DB00171 ) ) channels in pancreatic beta-cells , and its early insulin release and short duration of action would be effective in improving postprandial hyperglycemia . In studies of various cloned K( DB00171 ) channels , mitiglinide shows a higher selectivity for the beta-cell type of Q09428 /Kir6.2 than the cardiac and smooth muscle types of K( DB00171 ) channels in comparison with glibenclamide and glimepiride . In vitro and in vivo studies demonstrated the insulinotropic effect of mitiglinide is more potent than that of nateglinide , and mitiglinide surpassed in controlling postprandial hyperglycemia in normal and diabetic animals . In clinical trials , treatment with mitiglinide provided lasting improvement of postprandial hyperglycemia in Type 2 diabetic patients and decreased the fasting plasma glucose levels and HbA(1C) values . The incidence of adverse events related to mitiglinide were nearly equivalent to placebo ; in particular there was no difference with the frequency of hypoglycemia . The results from these studies indicated that mitiglinide could be expected to possess good therapeutic features of being effective in reducing postprandial glucose excursions in the early stage of Type 2 diabetes and less incidence of events suggestive of hypoglycemia . Attenuation of angiotensin II signaling recouples P29474 and inhibits nonendothelial NOX activity in diabetic mice . Angiotensin II ( Ang II ) levels are increased in patients with diabetes , but mechanisms underlying its contribution to diabetic vascular diseases are incompletely understood . We recently reported that in aortic endothelial cells , Ang II induces endothelial nitric oxide synthase ( P29474 ) uncoupling to produce superoxide ( O(2)*(-) ) rather than nitric oxide ( NO* ) , upon loss of the tetrahydrobiopterin ( H(4)B ) salvage enzyme dihydrofolate reductase ( P00374 ) . Here , we found that streptozotocin-induced diabetic mice had a marked increase in aortic O(2)*(-) production , which was inhibited by DB04223 methyl ester hydrochloride , indicating uncoupling of P29474 . Ang II receptor type 1 blocker candesartan or P12821 inhibitor captopril markedly attenuated P29474 -derived O(2)*(-) and hydrogen peroxide production while augmenting NO* bioavailability in diabetic aortas , implicating recoupling of P29474 . O(2)*(-) and NO* production were characteristically and quantitatively measured by electron spin resonance . P00374 expression was decreased in diabetic aortas but significantly restored by candesartan or captopril . Either also improved vascular H(4)B content and endothelium-dependent vasorelaxation in diabetes . Rac1-dependent NAD(P)H oxidase ( NOX ) activity was more than doubled in the endothelium-denuded diabetic aortas but was attenuated by candesartan or captopril , indicating that NOX remains active in nonendothelial vascular tissues , although uncoupled P29474 is responsible for endothelial production of O(2)*(-) . These data demonstrate a novel role of Ang II in diabetic uncoupling of P29474 and that Ang II-targeted therapy improves endothelial function via the novel mechanism of recoupling P29474 . Dual effectiveness on uncoupled P29474 and NOX may explain the high efficacy of Ang II antagonists in restoring endothelial function . P01308 secretory defects and impaired islet architecture in pancreatic beta-cell-specific P40763 knockout mice . Normal islet formation and function depends on the action of various growth factors operating in pre- and postnatal development ; however , the specific physiological function of each factor is largely unknown . Loss-of-function analyses in mice have provided little information so far , perhaps due to functional redundancies of the growth factors acting on the pancreas . The present study focuses on the role of the transcription factor P40763 in insulin-producing cells . P40763 is one of the potential downstream mediators for multiple growth factors acting on the pancreatic beta-cells , including betacellulin , hepatocyte growth factor , growth hormone , and heparin-binding P01133 -like growth factor . To elucidate its role in the beta-cells , the P40763 gene was disrupted in insulin-producing cells in mice ( P40763 -insKO ) , using a cre-mediated gene recombination approach . Unexpectedly , P40763 -insKO mice exhibited an increase in appetite and obesity at 8 weeks of age or older . The mice showed partial leptin resistance , suggesting that expression of the RIP ( rat insulin promoter ) -cre transgene in hypothalamus partially inhibited the appetite-regulating system . Intraperitoneal glucose tolerance tests , performed in non-obese 5-week-old mice , showed that the P40763 -insKO mice were glucose intolerant . Islet perifusion experiments further revealed a deficiency in early-phase insulin secretion . Whereas islet insulin content or islet mass was not affected , expression levels of P11168 , Q09428 , and P15692 were significantly reduced in P40763 -insKO islets . Interestingly , P40763 -insKO mice displayed impaired islet morphology : alpha-cells were frequently seen in central regions of islets . Our present observations demonstrate a unique role of P40763 in maintaining glucose-mediated early-phase insulin secretion and normal islet morphology . Renal lymph circulation blockage alters the epithelial cell phenotype and tubular integrity : role of distinct regulation of P18075 and TGF-β/Smads signaling pathway . AIMS : To investigate the effect of lymph circulation blockage on the alteration of renal epithelial cell phenotype and the tubular integrity , as well as the underlying mechanisms . METHODS : Wistar rats received left renal lymph ligation and right renal nephrectomy ( KL group ) or right renal nephrectomy without lymph ligation ( KN group ) and then were killed on day 14 , day 28 and day 56 . The urine , blood and kidney tissue were collected for the analysis of protein and gene expressions and morphological changes . RESULTS : The urine albumin and serum creatinine ( Cr ) in KL group were significantly increased compared with KN group . Masson and DB00233 staining indicated the epithelial cell degeneration , necrosis , sublethal loss and atrophy in KL rats , but not in KN group . Interestingly , from the atrophic tubules , some epithelial cells exhibited polarity changes with hypertrophy contrasting to the normal epithelial morphology of KN group throughout the experiment . By EM , ligated kidneys showed irregularly wrinkled basement membranes and epithelial cell swelling . Some intertubular areas of the KL kidney were expanded with fibrotic matrix and fibroblast-like cells . In line with these morphological changes , the fibroblast cell markers of FSP1 and α-SMA were markedly increased in contrast to the remarkable reduction in epithelial cell marker P12830 and tight junction protein ZO-1 . Moreover , the TGF-β1/ Q15796 /3 signaling pathway was significantly activated in KL rats in contrast to a robust downregulation of P18075 / Q99717 signaling . CONCLUSIONS : Disturbance of renal lymphatic circulation resulted in the epithelial cell phenotypic alteration and impaired tubular integrity possibly via distinct regulation of TGFβ1/Smads and P18075 / Q99717 signaling pathway . 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 . F-actin involvement in guinea pig sperm motility . Sperm motility is a must for natural fertilization to occur . During their travel through the epididymis , mammalian spermatozoa gradually acquire the ability to move . This is accomplished through a sliding movement of the outer doublet microtubules of the axoneme which is energized by the dynein ATPase . Within its complex structure , the mammalian sperm flagellum contains F-actin and thus , we decided to test in the guinea pig sperm flagellum the role of F-actin in motility . During maturation , capacitation , and the acrosome reaction , a gradual decrease of the relative concentration of F-actin was observed . Motility increased as spermatozoa became able to fertilize . P06396 , phalloidin , and KI inhibited sperm motility . P06396 canceled sperm motility within 20 min of treatment while 0.6 M KI had immediate effects . Phalloidin diminished hyperactive sperm motility slightly . All three compounds significantly increased the relative concentration of F-actin . Latrunculins are conventional drugs that destabilize the F-actin cytoskeleton . DB02621 ( O43561 A ) did not affect sperm motility ; but significantly increased F-actin relative concentration . The results suggested that in guinea pig spermatozoa , randomly severing F-actin filaments inhibits flagellar motility ; while end filament alteration does not . Thus , specific filament regions seem to be important for sperm motility . Could treatment with arundic acid ( DB05343 ) increase vulnerability for depression ? Arundic acid ( DB05343 ) is believed to be neuroprotective because of its actions on glia cells ; i.e. , its inhibitory effects on the synthesis of a calcium-binding protein P04271 . DB05343 is undergoing clinical trials for the treatment of patients with stroke and Alzheimer 's disease . Recent clinical studies point to a pervasive comorbidity of depression with stroke and Alzheimer 's disease . Previously , P04271 has been implicated in the pathobiological mechanisms of depression . Preclinical studies have shown that antidepressant treatment significantly increases brain P04271 . Here we hypothesize that available data that link P04271 with depression , along with the proposed inhibitory action of DB05343 on P04271 synthesis , indicate that this compound could increase vulnerability for depression in patients at risk for this disorder , and we propose that evaluation of patients with stroke and Alzheimer 's disease for the presence of depression should be routine in clinical trials employing DB05343 . Although it may be open for discussion whether the neuroprotective effects of DB05343 are exclusively due to its inhibition of P04271 synthesis , the latter action of DB05343 warrants studies of the effects of this drug in the pathobiology of depression . [ Vaccine-associated paralytic poliomyelitis showing biphasic motor paresis ] . We report a 38-year-old man with vaccine associated paralytic poliomyelitis ( VAPP ) which showed unusual biphasic worsening . The patient developed mild paresis of left upper and right lower extremities , five weeks after the oral poliovirus vaccination of patient 's son and two weeks after the intramuscular injection of mumps/varicella vaccine in the left triceps muscle for himself . Needle electromyography ( EMG ) of his left arm and right leg was not remarkable , and the weakness recovered almost completely in three weeks . However , four weeks after the needle EMG , severe weakness and muscle atrophy of the four extremities , accentuated at the left arm and right leg , developed again . Cervical Q9BWK5 showed gadolinium-enhanced , T(2) high-signal intensity area in the left C4- P13671 anterior horn , most prominent at the height of P01031 spine . Significant elevation of serum anti-poliomyelitis type 2 neutralizing antibody confirmed the diagnosis of VAPP . Immunomodulatory treatment , intravenous immunoglobulin ( DB00028 ) , did not improve weakness . We consider that the second clinical worsening of this patient was provoked by the needle EMG performed just after the first exacerbation , which injured the skeletal muscles and might have enhanced the retrograde transport of poliovirus via neural pathway . Targeting eIF4GI translation initiation factor affords an attractive therapeutic strategy in multiple myeloma . BACKGROUND : Deregulation of protein synthesis is integral to the malignant phenotype and translation initiation is the rate limiting stage . Therefore , eIF4F translation initiation complex components are attractive therapeutic targets . METHODS : Protein lysates of myeloma cells ( cell lines/patients ' bone marrow samples ) untreated/treated with bevacizumab were assayed for eIF4GI expression , regulation ( P15559 /proteosome dependent fragmentation ) ( WB , DB00266 , qPCR ) and targets (WB). eIF4GI was inhibited by knockdown and 4EGI-1 . Cells were tested for viability ( ELISA ) , death ( FACS ) and eIF4GI targets ( WB ) . RESULTS : Previously , we have shown that manipulation of P15692 in myeloma cells attenuated P06730 dependent translation initiation . Here we assessed the significance of eIF4GI to MM cells . We demonstrated increased expression of eIF4GI in myeloma cells and its attenuation upon P15692 inhibition attributed to elevated P15559 /proteasome dependent fragmentation and diminished mRNA levels . Knockdown of eIF4GI was deleterious to myeloma cells phenotype and expression of specific molecular targets ( Q99717 /ERα/HIF1α/c-Myc ) . Finally , we showed that the small molecule 4EGI-1 inhibits eIF4GI and causes a reduction in expression of its molecular targets in myeloma . CONCLUSION : Our findings substantiate that translation initiation of particular targets in MM is contingent on the function of eIF4GI , critical to cell phenotype , and mark it as a viable target for pharmacological intervention . Identification of epigenetically silenced genes in tumor endothelial cells . Tumor angiogenesis requires intricate regulation of gene expression in endothelial cells . We recently showed that DNA methyltransferase ( P26358 ) and histone deacetylase ( HDAC ) inhibitors directly repress endothelial cell growth and tumor angiogenesis , suggesting that epigenetic modifications mediated by DNMTs and HDAC are involved in regulation of endothelial cell gene expression during tumor angiogenesis . To understand the mechanisms behind the epigenetic regulation of tumor angiogenesis , we used microarray analysis to perform a comprehensive screen to identify genes down-regulated in tumor-conditioned versus quiescent endothelial cells , and reexpressed by DB01262 ( P22760 ) and trichostatin A ( P32119 ) . Among the 81 genes identified , 77 % harbored a promoter CpG island . Validation of mRNA levels of a subset of genes confirmed significant down-regulation in tumor-conditioned endothelial cells and reactivation by treatment with a combination of P22760 and P32119 , as well as by both compounds separately . Silencing of these genes in tumor-conditioned endothelial cells correlated with promoter histone H3 deacetylation and loss of H3 lysine 4 methylation , but did not involve DNA methylation of promoter CpG islands . For six genes , down-regulation in microdissected human tumor endothelium was confirmed . Functional validation by RNA interference revealed that clusterin , fibrillin 1 , and quiescin Q6 are negative regulators of endothelial cell growth and angiogenesis . In summary , our data identify novel angiogenesis-suppressing genes that become silenced in tumor-conditioned endothelial cells in association with promoter histone modifications and reactivated by P26358 and HDAC inhibitors through reversal of these epigenetic modifications , providing a mechanism for epigenetic regulation of tumor angiogenesis . The effect of aspirin and nonsteroidal anti-inflammatory drugs on prostaglandins . Cyclic prostanoids play important physiologic roles in inflammation and maintaining normal function of several organ systems . Prostaglandin production requires the conversion of arachidonate to the intermediate prostaglandin H2 , by the 2-step cyclo-oxygenation and peroxidation catalyzed by the enzyme cyclo-oxygenase ( also called prostaglandin H synthase ) . DB00945 and nonsteroidal anti-inflammatory drugs ( NSAIDs ) block the production of cyclic prostanoids by binding in different ways to this enzyme and blocking the active site . This results in decreased inflammation , but it can also produce side effects in the gastrointestinal tract , kidney , and platelets . Recent data demonstrate that there are 2 isoforms of the cyclo-oxygenase enzyme , called P23219 and P35354 . These isoforms are similar in size , substrate specificity , and kinetics , but vary in their expression and distribution . Normal physiologic functions appear to be maintained by P23219 , while P35354 appears to mediate the inflammatory response . Nonsteroidal drugs with selective inhibitory activity on the P35354 isoform should theoretically decrease inflammation while maintaining normal physiologic prostaglandin levels . Current NSAIDs are not selective enough to confirm this , but newer , more selective inhibitors of P35354 may answer this important question . Epigenetic regulation of protein tyrosine phosphatase Q05209 in triple-negative breast cancer . AIMS : The present study showed that the expression of Q05209 is epigenetically regulated . DB00928 ( 5-Azac ) , a DNA hypomethylating agent , significantly increased the expression of Q05209 at low concentrations ( 1μM and 2.5μM ) and decreased the expression of Q05209 at 5μM in the MDA-MB-231 and BT-549 triple-negative breast cancer cell lines . MAIN METHODS : Human MCF-7 , MDA-MB-231 and BT-549 cells were exposed to different concentrations of 5-Azac for 24 and 48h . RT-PCR was performed to determine the mRNA expression of Q05209 , P12830 and miRNA-124 . Western blotting was performed to assess the protein expression of various proteins , including Q05209 , P12830 , P26358 and PARP . KEY FINDINGS : 5-Azac , a DNA hypomethylating agent , significantly increased the expression of Q05209 at low concentrations ( 1μM and 2.5μM ) and decreased Q05209 expression at 5μM . We provide the first evidence that Q05209 expression is epigenetically regulated and that it is up-regulated at a lower dose of a P26358 inhibitor in MDA-MB-231 and BT-549 cells . Interestingly , the levels of miRNA-124 were increased only at 5μM , the concentration at which Q05209 expression was suppressed . SIGNIFICANCE : To the best of our knowledge , this is the first report that highlights the therapeutic potential of low-dose 5-Azac for the treatment of TNBC . Therefore , 5-Azac , an agent that has already been tested in acute myeloid leukemia , may be more effective at lower doses for the treatment of triple-negative breast cancer . Monoclonal antibody treatments for rheumatoid arthritis . INTRODUCTION : Rheumatoid arthritis ( RA ) is a systemic disease and the most prevalent of all autoimmune disorders . Here we review recent advances in the development and availability of biologic agents with a focus on monoclonal antibody or smaller formats of targeted engineered therapeutics including novel , non-antibody-based therapeutics . AREAS COVERED : Today an array of biologics blocking either proinflammatory cytokines or lymphocyte activation/survival are available that enable a substantial improvement over conventional disease-modifying antirheumatic drugs ( DMARDs ) . We review the engineering process of antibody-based biologics , their preclinical and clinical application , and current efforts to treat RA by interfering with B-cell function ( notable targets covered are P11836 , P28907 , B-cell activating factor , transmembrane activator and calcium-modulating and cyclophilin interactor ) , with T-cell function ( CD3 , P01730 , P10747 ) , with bone erosion ( O14788 ) , and with cytokines or growth factors ( tumor necrosis factor , interleukin-1 [ IL-1 ] , P05231 , Q16552 , P15692 ) . Future treatment choices might encompass the blockade or modulation of danger-associated molecular patterns such as P09429 , pattern recognition receptors , messenger RNAs or noncoding RNAs , histone acetylation , and inflammasome components . EXPERT OPINION : Although current therapies can reduce the signs and symptoms of RA for many patients , the quest for a cure ( or a more complete blockade of the structural damage ) in RA is still ongoing and will need treatment approaches , which are not exclusively confined to blocking a particular cytokine , receptor , or autoreactive B or T cell involved in disease progression . To this end exciting treatment alternatives and drug targets are on the horizon that may become available to patients in the future . DNA methyl transferase I acts as a negative regulator of allergic skin inflammation . The role of DNA methyl transferase I ( P26358 ) in allergic inflammation was investigated . Antigen stimulation decreased expression of P26358 in rat basophilic leukemia cells ( RBL2H3 ) . The down regulation of P26358 induced expression of histone deacetylase 3 ( O15379 ) . O15379 was necessary for allergic skin inflammation , such as such as triphasic cutaneous reaction and passive cutaneous anaphylaxis . The down regulation of P26358 resulted from activation of PKC and rac1 which were necessary for proteasome-dependent ubiquitination of P26358 by antigen stimulation . N-acetyl-L-cysteine , an inhibitor of reactive oxygen species production , exerted negative effects on allergic skin inflammation . Antigen stimulation led to increased expression of Q92993 , a histone acetyl transferase . Wild type , but not mutant form , Q92993 decreased expression of P26358 while increasing expression of O15379 , suggesting role for acetylation in ubiquitin-dependent proteasomal degradation of P26358 . In vivo down regulation of P26358 increased ear thickness , typical of allergic skin inflammation , induced vascular leakage and promoted angiogenesis in BALB/c mouse . The down regulation of P26358 enhanced angiogenic potential of rat aortic endothelial cells ( RAEC ) accompanied by activation of VEGR-2 and induced interaction between VEGR-2 and syk in RAEC . The enhanced angiogenic potential of RAEC was associated with the induction of P15692 by down regulation of P26358 in RBL2H3 cells . The down regulation of P26358 induced leukocytes-endothelial cell interaction and expression of various adhesion molecules . DB00945 exerted a negative effect on allergic skin inflammation by indirect regulation on P26358 via Q92993 . Taken together , these results suggest novel role for P26358 in allergic skin inflammation . DB09052 induces autologous T-cell killing of chronic lymphocytic leukemia cells . Chronic lymphocytic leukemia is an incurable B-cell malignancy that is associated with tumor cell-mediated T-cell dysfunction . It therefore represents a challenging disease for T-cell immunotherapeutics . The P15391 /CD3 bi-specific antibody construct blinatumomab ( AMG103 or MT103 ) has been tested clinically in non-Hodgkin 's lymphoma and acute lymphoblastic leukemia but has not been assessed in chronic lymphocytic leukemia . We investigated whether blinatumomab could overcome T-cell dysfunction in chronic lymphocytic leukemia in vitro . DB09052 was tested on peripheral blood mononuclear cells from 28 patients ( treatment naïve and previously treated ) . T-cell activation and function , as well as cytotoxicity against leukemic tumor cells were measured . DB09052 induced T-cell activation , proliferation , cytokine secretion and granzyme B release in a manner similar to that occurring with stimulation with anti-CD3/anti- P10747 beads . However , only blinatumomab was able to induce tumor cell death and this was found to require blinatumomab-mediated conjugate formation between T cells and tumor cells . Cytotoxicity of tumor cells was observed at very low T-cell:tumor cell ratios . A three-dimensional model based on confocal microscopy suggested that up to 11 tumor cells could cluster round each T cell . Importantly , blinatumomab induced cytotoxicity against tumor cells in samples from both treatment-naïve and treated patients , and in the presence of co-culture pro-survival signals . The potent cytotoxic action of blinatumomab on tumor cells appears to involve conjugation of T cells with tumor cells at both the activation and effector stages . The efficacy of blinatumomab in vitro suggests that the bi-specific antibody approach may be a powerful immunotherapeutic strategy in chronic lymphocytic leukemia . Epithelial mesenchymal transition during the neoplastic transformation of human breast epithelial cells by estrogen . Epithelial-mesenchymal transition ( EMT ) in epithelial cells has been indicated as an important component of neoplastic transformation although , the genetic mechanism involved in this process has not been defined . The aim of this study was to evaluate the expression of different genes related to EMT such as P12830 , TGFbeta1 , TGFbeta2 , h- DB01367 , Q15672 , SNAIL2 , Q99717 , P02751 , P13688 and P78504 using the in vitro-in vivo model of the estrogen induced cell transformation developed in our laboratory . The E2-transformed MCF-10F ( E2 70 ) cells and the tumorigenic cell line P01031 - Q92854 -T8 ( P01031 -T8 ) exhibit progressive loss of ductulogenesis as demonstrated by growth in collagen matrix . MCF-10F cells form ductal structures while E2 70 cells form solid spherical masses that in histological sections exhibit a pattern of growth resembling ductal hyperplasia or carcinoma in situ . The tumorigenic cells P01031 -T8 did not form structures on collagen acquiring an invasive pattern with spindle like features . We have observed a reduction in P12830 expression in E2 70 cells and a complete loss in P01031 -T8 cells . TGFbeta1 , TGFbeta2 , P13688 and P78504 were down-regulated in E2 70 and P01031 -T8 cells . Q99717 and h- DB01367 were up-regulated in the tumorigenic P01031 -T8 cells whereas P02751 , Twist1 and Snail2 were up-regulated in P01031 -T8 and down-regulated in E2 70 . We conclude that the loss of expression of TGFbeta1 , TGFbeta2 , P13688 and P78504 are related to ductulogenesis and branching and the overexpression of h- DB01367 with loss of P12830 expression and up-modulation of Q15672 , SNAIL2 and Q99717 expressions are involved in the EMT modulation .

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P15056 inhibitors suppress apoptosis through off-target inhibition of JNK signaling . DB08881 and dabrafenib selectively inhibit the P15056 ( P15056 ) kinase , resulting in high response rates and increased survival in melanoma . Approximately 22 % of individuals treated with vemurafenib develop cutaneous squamous cell carcinoma ( cSCC ) during therapy . The prevailing explanation for this is drug-induced paradoxical P29323 activation , resulting in hyperproliferation . Here we show an unexpected and novel effect of vemurafenib/PLX4720 in suppressing apoptosis through the inhibition of multiple off-target kinases upstream of c-Jun N-terminal kinase ( JNK ) , principally Q9NYL2 . JNK signaling is suppressed in multiple contexts , including in cSCC of vemurafenib-treated patients , as well as in mice . Expression of a mutant Q9NYL2 that can not be inhibited reverses the suppression of JNK activation and apoptosis . Our results implicate suppression of JNK-dependent apoptosis as a significant , independent mechanism that cooperates with paradoxical P29323 activation to induce cSCC , suggesting broad implications for understanding toxicities associated with P15056 inhibitors and for their use in combination therapies . DOI : http://dx.doi.org/10.7554/eLife.00969.001 . DB01269 : a new anti- P00533 antibody for the treatment of advanced colorectal cancer . Neuroprotection mediated via neurotrophic factors and induction of neurotrophic factors . Neurotrophins and other neurotrophic factors have been shown to support the survival and differentiation of many neuronal populations of the central and peripheral nervous system . Therefore , administering neurotrophic factors could represent an alternative strategy for the treatment of acute and chronic brain disorders . However , the delivery of neurotrophic factors to the brain is one of the largest obstacles in the development of effective therapy for neurodegenerative disorders , because these proteins are not able to cross the blood-brain barrier . The induction of growth factor synthesis in the brain tissue by systemically administered lipophilic drugs , such as beta-adrenoceptor agonists , shown to increase endogenous nerve growth factor ( P01138 ) synthesis in the brain , would be an elegant way to overcome these problems of application . Stimulation of beta-adrenoceptors with clenbuterol led to increased P01138 synthesis in cultured central nervous system ( CNS ) cells and rat brain tissue . DB01407 -induced P01138 expression was reduced to the control levels by coadministration of beta-adrenoceptor antagonist propranolol . Furthermore , clenbuterol protected rat hippocampal neurons subjected to excitotoxic damage . The neuroprotective effect of clenbuterol in vitro depended on increased P01138 synthesis , since the neuroprotection was abolished by P01138 antisense oligonucleotide as well as by antibodies directed against P01138 itself . In vivo , clenbuterol protected rat hippocampus in a model of transient forebrain ischemia and reduced the infarct volume in a rat model of permanent middle cerebral artery occlusion ( MCAo ) . The neuroprotective effect of clenbuterol in vivo was accompanied by enhanced P01138 synthesis in brain tissue . These findings support our hypothesis that orally active P01138 inducers may have a potential as therapeutic agents for the treatment of neurodegenerative disorders and stroke . P35372 phosphorylation , desensitization , and ligand efficacy . Mu opioid receptors are subject to phosphorylation and desensitization through actions of at least two distinct biochemical pathways : agonist-dependent mu receptor phosphorylation and desensitization induced by a biochemically distinct second pathway dependent on protein kinase C activation ( 1 ) . To better understand the nature of the agonist-induced mu receptor phosphorylation events , we have investigated the effects of a variety of opiate ligands of varying potencies and intrinsic activities on mu receptor phosphorylation and desensitization . Exposure to the potent full agonists sufentanil , dihydroetorphine , etorphine , etonitazine , and [ D-Ala2 , MePhe4 , Glyol5 ] enkephalin ( DAMGO ) led to strong receptor phosphorylation , while methadone , l-alpha-acetylmethadone ( DB01227 ) , morphine , meperidine , DADL , beta-endorphin(1-31) , enkephalins , and dynorphin A(1-17) produced intermediate effects . The partial agonist buprenorphine minimally enhanced receptor phosphorylation while antagonists failed to alter phosphorylation . DB00921 and full antagonists each antagonized the enhanced mu receptor phosphorylation induced by morphine or DAMGO . The rank order of opiate ligand efficacies in producing mu receptor-mediated functional desensitization generally paralleled their rank order of efficacies in producing receptor phosphorylation . Interestingly , the desensitization and phosphorylation mediated by methadone and DB01227 were disproportionate to their efficacies in two distinct test systems . This generally good fit between the efficacies of opiates in mu receptor activation , phosphorylation , and desensitization supports the idea that activated receptor/agonist/G-protein complexes and/or receptor conformational changes induced by agonists are required for agonist-induced mu receptor phosphorylation . Data for methadone and DB01227 suggest possible contribution from their enhanced desensitizing abilities to their therapeutic efficacies . The recovery of the B-cell population in adult thymectomized , lethally irradiated and bone marrow-reconstituted mice . The recovery of the B-cell population in adult thymectomized , irradiated and bone marrow-reconstituted mice ( T X BM mice was estimated at various times after bone marrow transplantation . The spleen cells to be tested were mixed with dexamethasone-resistant thymocytes ( P29323 ) and sheep red blood cells ( SRBC ) and transferrred to irradiated recipients . The number of plaque-forming cells ( P27918 ) in the spleen of the recipients was determined 7 days later . Using this functional B-cell assay a sequential appearance of the precursors of IgM- , IgG- and IgA- P27918 in the spleen of T X BM mice was observed . The precursors of IgM-PFG ( IgM-B cells ) were present immediately after transplantation . The first IgG-B cells could be detected at 13-16 days after transplantation and the IgA-B cells finally appeared at 22 days after transplantation . The number of B cells reached a constant and normal level at 30 days after transplantation . The IgM- , IgG- and IgA-B cell development in sham-thymectomized , irradiated and bone narrow-reconstituted mice ( ST X BM mice ) was virtually the same as in T X BM mice . Q8N0V5 V ( Mgat5 ) -mediated N-glycosylation negatively regulates Th1 cytokine production by T cells . The differentiation of naive P01730 (+) T cells into either proinflammatory Th1 or proallergic Th2 cells strongly influences autoimmunity , allergy , and tumor immune surveillance . We previously demonstrated that beta1,6GlcNAc-branched complex-type ( Q8N0V5 V ( Mgat5 ) ) N-glycans on TCR are bound to galectins , an interaction that reduces TCR signaling by opposing agonist-induced TCR clustering at the immune synapse . Mgat5(-/-) mice display late-onset spontaneous autoimmune disease and enhanced resistance to tumor progression and metastasis . In this study we examined the role of beta1,6GlcNAc N-glycan expression in Th1/Th2 cytokine production and differentiation . beta1,6GlcNAc N-glycan expression is enhanced by TCR stimulation independent of cell division and declines at the end of the stimulation cycle . DB00075 -activated splenocytes and naive T cells from Mgat5(-/-) mice produce more P01579 and less P05112 compared with wild-type cells , the latter resulting in the loss of P05112 -dependent down-regulation of IL-4Ralpha . DB02034 , an inhibitor of Q16706 , blocked beta1,6GlcNAc N-glycan expression and caused a similar increase in P01579 production by T cells from humans and mice , but no additional enhancement in Mgat5(-/-) T cells . Mgat5 deficiency did not alter P01579 / P05112 production by polarized Th1 cells , but caused an approximately 10-fold increase in P01579 production by polarized Th2 cells . These data indicate that negative regulation of TCR signaling by beta1,6GlcNAc N-glycans promotes development of Th2 over Th1 responses , enhances polarization of Th2 cells , and suggests a mechanism for the increased autoimmune disease susceptibility observed in Mgat5(-/-) mice . Dermatological adverse events from P15056 inhibitors : a growing problem . The development of targeted therapies has ushered in a new era in the management of melanoma . Inhibitors of the DB01367 -RAF-MEK- P29323 pathway have taken the center stage with development at a rapid pace . DB08881 was recently approved by regulatory agencies , and other agents ( e.g. dabrafenib ) are in various stages of clinical testing . These agents are producing remarkable results for patients , but are also presenting new challenges . Clinical toxicities and drug resistance are topmost issues . Some of the most common and vivid representations of adverse events to these agents are the dermatologic manifestations . Published trials and initial observations reflect a toxicity profile ( e.g. squamous cell carcinomas/keratoacanthomas , maculopapular rashes , hyperkeratosis ) that is distinct from cutaneous toxicities from P00533 and P42345 inhibitors ( acneiform rash , paronychia , xerosis ) . Their management extends beyond conservative treatment and includes specific physical and surgical treatment modalities , skill sets unique to dermatologists . All these pose significant challenges to clinicians , and sound knowledge of such toxicities and their management will likely result in improved patient outcomes and quality of life . In this manuscript , we provide an overview of the emerging scientific literature on dermatological adverse events arising out of P15056 inhibition . Cardiac glycosides regulate endothelial tissue factor expression in culture . BACKGROUND : P13726 ( TF ) plays an important role in acute coronary syndromes and stent thrombosis . This study investigates whether Na(+)/K(+)-ATPase regulates TF expression in human endothelial cells . METHODS AND RESULTS : Ouabain inhibited tumor necrosis factor ( P01375 ) -alpha-induced endothelial TF protein expression ; maximal inhibition occurred at 10(-5) mol/L , reached more than 70 % , and was observed throughout the 5 hours stimulation period . The decrease in protein expression was paralleled by a reduced TF surface activity . Similarly , lowering of extracellular potassium concentration inhibited P01375 -induced TF protein expression . In contrast , ouabain did not affect P01375 -induced expression of full-length TF mRNA for up to 5 hours of stimulation ; instead , expression of alternatively-spliced TF mRNA was upregulated after 3 and 5 hours of stimulation . Ouabain did not affect P01375 -induced activation of the Q96HU1 kinases p38 , extracellular signal-regulated kinase ( P29323 ) , and c-Jun terminal NH(2) kinase ; activation of Akt and p70S6 kinase remained unaltered as well . Similar to the Q96HU1 kinases , ouabain did not affect P01375 -induced degradation of IkappaB-alpha . Ouabain had no effect on TF protein degradation . CONCLUSIONS : Na(+)/K(+)-ATPase is required for protein translation of endothelial TF in culture . This observation provides novel insights into posttranscriptional regulation of TF expression . Effective dasatinib uptake may occur without human organic cation transporter 1 ( O15245 ) : implications for the treatment of imatinib-resistant chronic myeloid leukemia . We have previously shown that imatinib uptake into chronic myeloid leukemia ( CML ) cells is dependent on human organic cation transporter 1 ( O15245 ; O15245 ) , and that low O15245 expression is an important determinant of clinical outcome to imatinib treatment . We hypothesized that dasatinib might be transported differently than imatinib , possibly accounting for its favorable effects in imatinib-resistant patients . (14)C-dasatinib uptake was greater in KCL22-transfected cells with pcDNA3- O15245 plasmid ( high O15245 -expressing cells ) than in control cells ( P = .02 ) . However , hOCT inhibitors did not decrease dasatinib uptake into either control or primary cells , in contrast to their block on imatinib uptake . Dasa-tinib decreased the level of phosphorylated CrkL to 49.9 % in control and 40.3 % in high O15245 -expressing cells . Dasa-tinib efflux was investigated in confluent P08183 -transfected MDCKII cell monolayers . Both dasatinib and imatinib were transported from the basal to the apical layer , indicating that they were transported by P08183 , which was confirmed using the P08183 inhibitor PSC833 ( P = .001 and P < .001 , respectively ) . Compared with imatinib , dasatinib achieved superior intracellular levels and P11274 - P00519 suppression even in cells with low or blocked O15245 . Efflux of dasatinib and imatinib appear similar via P08183 . Dasatinib may therefore offer an advantage over imatinib in patients with low O15245 expression . mu-Opioid receptor agonists differentially regulate the expression of miR-190 and Q13562 . The agonists of mu-opioid receptor ( P35372 ) induce extracellular signal-regulated kinase ( P29323 ) phosphorylation through different pathways : morphine uses the protein kinase C ( PKC ) -pathway , whereas fentanyl functions in a beta-arrestin2-dependent manner . In addition , the two pathways result in the different cellular location of phosphorylated P29323 and the activation of different sets of transcriptional factors . In the current study , the influence of the two pathways on the expression of microRNAs ( miRNAs ) was investigated . After treating the primary culture of rat hippocampal neurons and the mouse hippocampi with morphine or fentanyl for 3 days , seven miRNAs regulated by one or two of the agonists were identified . One of the identified miRNAs , miR-190 , was down-regulated by fentanyl but not by morphine . This down-regulation was attenuated by 1,4-diamino-2,3-dicyano-1,4-bis(methylthio)butadiene ( U0126 ) , which blocks the phosphorylation of P29323 . When fentanyl-induced but not morphine-induced P29323 phosphorylation was blocked in the primary cultures from beta-arrestin2(-/-) mouse , fentanyl did not decrease the expression of miR-190 . However , a PKC inhibitor that blocked morphine-induced P29323 phosphorylation specifically had no effect on the miR-190 down-regulation . Therefore the decrease in miR-190 expression resulted from the agonist-selective P29323 phosphorylation . In addition , the expressional changes in one of the miR-190 targets , neurogenic differentiation 1 ( Q13562 ) , correlated with those in miR-190 expression , suggesting the P35372 could regulate the Q13562 pathways via the control of miR-190 expression . DB08881 for the treatment of P15056 mutant metastatic melanoma . DB08881 was the first selective P15056 inhibitor licensed in cancer . It is indicated for the treatment of patients affected by advanced melanoma with P15056 V600 mutation . It has shown successful results in terms of efficacy together with a favorable toxicity profile . Other compounds such as the P15056 inhibitor dabrafenib and the immunotherapeutic agent ipilimumab are also approved in the same group of patients . This article reviews the chemistry , pharmacokinetics , pharmacodynamics and clinical development of vemurafenib . Moreover , its efficacy and toxicity are compared with dabrafenib and ipilimumab . A number of trials with vemurafenib alone or in combination with other drugs are also analyzed . These trials will determine the role of vemurafenib in the treatment of P15056 mutant melanoma in forthcoming years . DB09045 for the treatment of type 2 diabetes . DB09045 is a novel glucagon-like peptide 1 ( P0C6A0 ) receptor agonist with a unique structure that supports once-weekly dosing in patients with type 2 diabetes ( T2DM ) , most of whom have a big pill burden . It appears to be efficacious in reducing hemoglobin A1c ( HbA1c ) up to 1.59 % and promotes modest weight loss up to 3 kg with a low incidence of hypoglycemia and mild to moderate gastrointestinal adverse events . Convenient weekly dosing could improve compliance and help attain sustained glycemic goals . Addressing obesity is an integral part of T2DM management and weight loss may contribute to better glycemic and cardiovascular benefits . Results of ongoing clinical trials on cardiovascular safety are important to determine the risk-to-benefit ratio . As with any drug , patient selection and ongoing monitoring will be important . If approved , dulaglutide will be one of the first weekly P43220 agonist to be available in a ready-to-use pen device with an automatic injector . Immunologic effects of an orally available BRAFV600E inhibitor in P15056 wild-type murine models . DB08881 is an orally available small molecule that targets constitutively activated BRAFV600E , an integral part of the MAPK pathway involved in melanomagenesis . We examined the effects of vemurafenib on cytokine production and antitumor response in a P15056 wild-type ( WT ) non-tumor-bearing murine model and a P15056 WT murine insulinoma system to determine its effect on immune function during immunotherapy . We demonstrate no significant effect from vemurafenib on P01730 + and CD8+ T-cell cytokine production or on a T-cell-mediated antitumor response . Our data demonstrate that vemurafenib does not significantly affect P15056 WT targets , suggesting that there may be a role for combining vemurafenib treatment with T-cell-directed immunotherapy . 3,4-methylenedioxymethamphetamine self-administration is abolished in serotonin transporter knockout mice . BACKGROUND : The neurobiological mechanism underlying the reinforcing effects of 3,4-methylenedioxymethamphetamine ( DB01454 ) remains unclear . The aim of the present study was to determine the contribution of the serotonin transporter ( P31645 ) in DB01454 self-administration behavior by using knockout ( KO ) mice deficient in P31645 . METHODS : Knockout mice and wild-type ( WT ) littermates were trained to acquire intravenous self-administration of DB01454 ( 0 , .03 , .06 , .125 , and .25 mg/kg/infusion ) on a fixed ratio 1 ( FR1 ) schedule of reinforcement . Additional groups of mice were trained to obtain food and water to rule out operant responding impairments . Microdialysis studies were performed to evaluate dopamine ( DA ) and serotonin ( 5-HT ) extracellular levels in the nucleus accumbens ( Q9C000 ) and prefrontal cortex ( P27918 ) , respectively , after acute DB01454 ( 10 mg/kg ) . RESULTS : None of the DB01454 doses tested maintained intravenous self-administration in KO animals , whereas WT mice acquired responding for DB01454 . Acquisition of operant responding for food and water was delayed in KO mice , but no differences between genotypes were observed on the last day of training . DB01454 increased DA extracellular levels to a similar extent in the Q9C000 of WT and KO mice . Conversely , extracellular concentrations of 5-HT in the P27918 were increased following DB01454 only in WT mice . CONCLUSIONS : These findings provide evidence for the specific involvement of P31645 in DB01454 reinforcing properties . Antineoplastic mechanisms of niclosamide in acute myelogenous leukemia stem cells : inactivation of the NF-kappaB pathway and generation of reactive oxygen species . NF-kappaB may be a potential therapeutic target for acute myelogenous leukemia ( AML ) because NF-kappaB activation is found in primitive human AML blast cells . In this report , we initially discovered that the potent antineoplastic effect of niclosamide , a Food and Drug Administration-approved antihelminthic agent , was through inhibition of the NF-kappaB pathway in AML cells . DB06803 inhibited the transcription and DNA binding of NF-kappaB . It blocked tumor necrosis factor-induced P25963 phosphorylation , translocation of p65 , and expression of NF-kappaB-regulated genes . DB06803 inhibited the steps TAK1 --> O15111 ( IKK ) and IKK --> P25963 . DB06803 also increased the levels of reactive oxygen species ( ROS ) in AML cells . Quenching ROS by the glutathione precursor DB06151 attenuated niclosamide-induced apoptosis . Our results together suggest that niclosamide inhibited the NF-kappaB pathway and increased ROS levels to induce apoptosis in AML cells . On translational study of the efficacy of niclosamide against AML , niclosamide killed progenitor/stem cells from AML patients but spared those from normal bone marrow . DB06803 was synergistic with the frontline chemotherapeutic agents cytarabine , etoposide , and daunorubicin . It potently inhibited the growth of AML cells in vitro and in nude mice . Our results support further investigation of niclosamide in clinical trials of AML patients . Enhancement of cytotoxicity of natural product drugs against multidrug resistant variant cell lines of human head and neck squamous cell carcinoma and breast carcinoma by tesmilifene . N,N-diethyl-2-[4-(phenylmethyl)phenoxyl]ethanamine ( tesmilifene ) , a tamoxifen derivative with antihistamine activity , greatly enhanced the survival of doxorubicin-treated , advanced stage breast cancer patients in a phase III trial . However , the molecular basis of tesmilifene action is not firmly established . The effects of tesmilifene on activity of several anticancer drugs was investigated using human head and neck squamous cell carcinoma ( HNSCC ) and breast carcinoma cell lines as a model system . Multidrug resistant ( MDR ) variants of an HNSCC cell line , HN-5a/V15e , and a breast carcinoma cell line , MCF-7/V25a , both highly overexpressed mdr1 ( P08183 ) mRNA and the proteins P-glycoprotein and glutathione transferase-pi . Drug sensitivities were measured by a vital stain after 4 days of continuous exposure to anticancer drug in the absence and presence of tesmilifene at a concentration that alone had no antiproliferative effect . DB04905 had minimal effect on drug cytotoxicity against the parental cell lines . However , the same tesmilifene treatment enhanced cytotoxicity of docetaxel , paclitaxel , epirubicin , doxorubicin , and vinorelbine against both MDR cell lines by up to 50 % . Flow cytometric measurement of annexin V/propidium iodide staining demonstrated that tesmilifene increased the killing of HN-5a/V15e cells caused by docetaxel after 24 and 48h exposure . DB04905 increased accumulation of radiolabelled vincristine in HN-5a/V15e cells , over 4h , by up to 100 % . The results suggest that tesmilifene might be effective in the treatment of tumors that are resistant to natural product drugs . The mechanism of enhancement appears to be related to expression of an ABC pump-dependent , MDR phenotype . P13726 and factor VIIa as therapeutic targets in disorders of hemostasis . For hemophilia patients with inhibitors against FVIII or FIX , the development of recombinant factor VIIa ( DB00036 ) raises the possibility of a therapeutic alternative whose availability and convenience of treatment are comparable to those of FVIII or FIX . In support of this new concept for the treatment of bleeding episodes , pharmacological doses of FVIIa have been shown to induce hemostasis . Pharmacological doses of DB00036 enhance thrombin generation on thrombin-activated platelets , thereby facilitating the formation of strong , well-structured fibrin plugs resistant to premature proteolysis . Modified DB00036 molecules with a stronger hemostatic potential have been produced . Inhibition of the FVII-TF-dependent pathway ( P10646 and rFVIIai ) has been tried in attempts to prevent thrombosis , with promising results in animal models so far not confirmed in clinical trials . 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 . Global expression analysis identified a preferentially nerve growth factor-induced transcriptional program regulated by sustained mitogen-activated protein kinase/extracellular signal-regulated kinase ( P29323 ) and AP-1 protein activation during PC12 cell differentiation . Neuronal differentiation of PC12 cells in response to P01138 is a prototypical model in which signal duration determines a biological response . Sustained P29323 activity induced by P01138 , as compared with transient activity induced by P01133 , is critical to the differentiation of these cells . To characterize the transcriptional program activated preferentially by P01138 , we compared global gene expression profiles between cells treated with P01138 and P01133 for 2-4 h , when sustained P29323 signaling in response to P01138 is most distinct from the transient signal elicited by P01133 . This analysis identified 69 genes that were preferentially up-regulated in response to P01138 . As expected , up-regulation of these genes was mediated by sustained P29323 signaling . In addition , they were up-regulated in response to other neuritogenic treatments ( pituitary adenylate cyclase-activating polypeptide and 12-O-tetradecanoylphorbol-13-acetate plus dbcAMP ) and were enriched for genes related to neuronal differentiation/function . Computational analysis and chromatin immunoprecipitation identified binding of CREB and AP-1 family members ( Fos , FosB , Fra1 , JunB , JunD ) upstream of > 30 and 50 % , respectively , of the preferentially P01138 -induced genes . Expression of several AP-1 family members was induced by both P01133 and P01138 , but their induction was more robust and sustained in response to P01138 . The binding of Fos family members to their target genes was similarly sustained in response to P01138 and was reduced upon MEK inhibition , suggesting that AP-1 contributes significantly to the P01138 transcriptional program . Interestingly , Fra1 as well as two other P01138 -induced AP-1 targets ( HB- P01133 and miR-21 ) function in positive feedback loops that may contribute to sustained AP-1 activity . Impaired MEK signaling and SERCA expression promote ER stress and apoptosis in insulin-resistant macrophages and are reversed by exenatide treatment . Accumulation of toxic lipids evokes the unfolded protein response ( UPR ) and apoptotic death of macrophages and vascular cells in atherosclerotic plaques . Primary macrophages from insulin-resistant ob/ob and insulin receptor ( Insr ) (-/-) mice display increased apoptosis in response to loading with free cholesterol or oxysterol , but underlying mechanisms have not been elucidated . We show increased activation of all three major branches of the UPR in response to free cholesterol or oxysterol loading in insulin-resistant macrophages . Inhibition and rescue experiments revealed that defective MEK/extracellular signal\x{2013}related kinase ( P29323 ) / DB02527 -responsive element-binding protein ( CREBP ) signaling in insulin-resistant macrophages leads to decreased expression of sarcoplasmic endoplasmic reticulum ( ER ) Ca(2+)-ATPase , depletion of ER calcium stores , P19525 -like ER kinase activation , and ER stress-associated apoptosis . Activation of macrophage glucagon-like peptide 1 ( P0C6A0 ) receptor via the antidiabetic drug exenatide led to improvements in both P29323 and AKT signaling and reversed the increase in UPR and apoptosis of insulin-resistant macrophages in atherosclerotic lesions of ob/ob.Ldlr(-/-) and Insr(-/-).Ldlr(-/-) mice . Increased signaling via P43220 or the CREBP activator protein kinase A thus offers a way to rescue insulin-resistant macrophages from excessive ER stress responses and apoptosis in insulin resistance and type 2 diabetes . Personalized medicine and pharmacogenetic biomarkers : progress in molecular oncology testing . In the field of oncology , clinical molecular diagnostics and biomarker discoveries are constantly advancing as the intricate molecular mechanisms that transform a normal cell into an aberrant state in concert with the dysregulation of alternative complementary pathways are increasingly understood . Progress in biomarker technology , coupled with the companion clinical diagnostic laboratory tests , continue to advance this field , where individualized and customized treatment appropriate for each individual patient define the standard of care . Here , we discuss the current commonly used predictive pharmacogenetic biomarkers in clinical oncology molecular testing : P15056 V600E for vemurafenib in melanoma ; Q9HC35 - Q9UM73 for crizotinib and P00533 for erlotinib and gefitinib in non-small-cell lung cancer ; P01116 against the use of cetuximab and panitumumab in colorectal cancer ; P04626 ( P04626 /neu ) for trastuzumab in breast cancer ; P11274 - P00519 for tyrosine kinase inhibitors in chronic myeloid leukemia ; and P29590 /RARα for all-trans-retinoic acid and arsenic trioxide treatment for acute promyelocytic leukemia .

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Persistent Cdk2 inactivation drives growth arrest of P11274 - P00519 -expressing cells in response to dual inhibitor of P12931 and P00519 kinases SKI606 . Complementary inhibition of tyrosine and P12931 kinases implement dual P12931 / P00519 inhibitor effects in chronic myeloid leukemia ( CML ) . Here , we show that one such inhibitor , DB06616 , induces persistent Cdk2 inactivation leading to growth arrest of P11274 - P00519 -expressing cells either IM-sensitive or driven to IM-resistance by other events than gene overexpression and point mutations . Inhibition of Akt serine/threonine kinase , a phosphatidylinositol 3 kinase ( PI-3k ) target that integrates Q92817 TK signaling with membrane-associated P12931 kinases , is a central component of restored expression and subcellular redistribution of Cdk2 regulatory signals ( P38936 and p27 and Cdc25A phosphatase ) in response to DB06616 . The putative roles of growth factor ( namely P08700 ) autocrine loop in P11274 - P00519 -expressing progenitor progression towards a drug-resistant phenotype are discussed . Neuroprotective effects of huperzine A . A natural cholinesterase inhibitor for the treatment of Alzheimer 's disease . DB04864 ( HupA ) , isolated from Chinese herb Huperzia serrata , is a potent , highly specific and reversible inhibitor of acetylcholinesterase . It has been found to reverse or attenuate cognitive deficits in a broad range of animal models . Clinical trials in China have demonstrated that HupA significantly relieves memory deficits in aged subjects , patients with benign senescent forgetfulness , Alzheimer 's disease ( AD ) and vascular dementia ( VD ) , with minimal peripheral cholinergic side effects compared with other AChEIs in use . HupA possesses the ability to protect cells against hydrogen peroxide , beta-amyloid protein ( or peptide ) , glutamate , ischemia and staurosporine-induced cytotoxicity and apoptosis . These protective effects are related to its ability to attenuate oxidative stress , regulate the expression of apoptotic proteins Bcl-2 , Bax , P04637 and caspase-3 , protect mitochondria , and interfere with P05067 metabolism . Antagonizing effects on DB01221 receptors and potassium currents may contribute to the neuroprotection as well . It is also possible that the non-catalytic function of P22303 is involved in neuroprotective effects of HupA . The therapeutic effects of HupA on AD or VD are probably exerted via a multi-target mechanism . P62937 and calcineurin functions investigated by gene inactivation , cyclosporin A inhibition and cDNA arrays approaches in the phytopathogenic fungus Botrytis cinerea . Calcineurin phosphatase and cyclophilin A are cellular components involved in fungal morphogenesis and virulence . Their roles were investigated in the phytopathogenic fungus Botrytis cinerea using gene inactivation , drug inhibition and cDNA macroarrays approaches . First , the BCP1 gene coding for cyclophilin A was identified and inactivated by homologous recombination . The bcp1Delta null mutant obtained was still able to develop infection structures but was altered in symptom development on bean and tomato leaves . Opposite to this , calcineurin inhibition using cyclosporin A ( DB00091 ) modified hyphal morphology and prevented infection structure formation . DB00091 drug pattern signature on macroarrays allowed the identification of 18 calcineurin-dependent ( CND ) genes among 2839 B. cinerea genes . Among the co-regulated CND genes , three were shown to be organized as a physical cluster that could be involved in secondary metabolism . The signature of BCP1 inactivation on macroarrays allowed the identification of only three BCP1 cyclophilin-dependent ( O75976 ) genes that were different from CND genes . Finally , no DB00091 drug pattern signature was observed in the bcp1Delta null mutant which provided a molecular target validation of the drug . Crystal structure of the O00206 -MD-2 complex with bound endotoxin antagonist DB04933 . O00206 and MD-2 form a heterodimer that recognizes LPS ( lipopolysaccharide ) from Gram-negative bacteria . DB04933 is an analog of LPS that antagonizes its activity by binding to the O00206 -MD-2 complex . We determined the structure of the full-length ectodomain of the mouse O00206 and MD-2 complex . We also produced a series of hybrids of human O00206 and hagfish VLR and determined their structures with and without bound MD-2 and DB04933 . O00206 is an atypical member of the LRR family and is composed of N-terminal , central , and C-terminal domains . The beta sheet of the central domain shows unusually small radii and large twist angles . MD-2 binds to the concave surface of the N-terminal and central domains . The interaction with DB04933 is mediated by a hydrophobic internal pocket in MD-2 . Based on structural analysis and mutagenesis experiments on MD-2 and O00206 , we propose a model of O00206 -MD-2 dimerization induced by LPS . BMI‑1 is important in bufalin‑induced apoptosis of K562 cells . The purpose of this study was to analyze the effects of bufalin on the gene expression of K562 cells and on the expression of BMI‑1 pathway constituents in K562 cell apoptosis . K562 cells were treated with bufalin , and the inhibition rate and apoptosis were detected by an MTT assay , flow cytometry and a microarray assay . BMI‑1 , p16INK4a and Q8N726 were examined by quantitative polymerase chain reaction ( qPCR ) . Bufalin induced significant changes in the gene expression of the K562 cells ; 4296 genes were differentially expressed , 2185 were upregulated and 2111 were downregulated . The most upregulated genes were associated with transcription regulation , while the most downregulated genes were associated with the non-coding RNA metabolic processes and DNA repair . qPCR analysis demonstrated that BMI‑1 was overexpressed in the K562 cells . Bufalin is able to downregulate BMI‑1 expression levels in K562 cells prematurely and cause an increase in the expression levels of p16INK4a and Q8N726 . Moreover , bufalin downregulated P11274 / P00519 expression levels in a time‑dependent manner , and the expression of P11274 / P00519 was not associated with the upregulation or downregulation of BMI‑1 expression . Bufalin may induce K562 cell apoptosis by downregulating BMI‑1 expression levels and accordingly upregulating the expression levels of p16INK4a and Q8N726 . Bufalin may also induce K562 cell apoptosis via downregulating P11274 / P00519 expression levels , and this pathway may be independent of the BMI‑1 pathway . Synergism between bosutinib ( DB06616 ) and the Chk1 inhibitor ( PF-00477736 ) in highly imatinib-resistant P11274 /ABL⁺ leukemia cells . Interactions between the dual P11274 / P00519 and Src inhibitor bosutinib and the Chk1 inhibitor PF-00477736 were examined in P11274 / P00519 (+) leukemia cells , particularly imatinib-resistant cells , including those with the T315I mutation . Bosutinib blocked PF-00477736-induced P27361 /2 activation and sharply increased apoptosis in association with Mcl-1 inhibition , p34(cdc2) dephosphorylation , BimEL up-regulation , and DNA damage in imatinib-resistant CML or Ph(+) ALL cell lines . Inhibition of Src or Q02750 by shRNA significantly enhanced PF-0047736 lethality . Bosutinib/PF-00477736 co-treatment also potentiated cell death in P28906 (+) CML patient samples , including dasatinib-resistant blast crisis cells exhibiting both T315I and E355G mutations , but was minimally toxic to normal P28906 (+) cells . Finally , combined in vivo treatment significantly suppressed BaF3/T315I tumor growth and prolonged survival in an allogeneic mouse model . Together , these findings suggest that this targeted combination strategy warrants attention in IM-resistant CML or Ph(+) ALL . [ Clinical and cytogenetic analyses of 45 adult patients with acute lymphoblastic leukemia ] . OBJECTIVE : To analyze the correlation between clinical features and cytogenetic finding of 45 adult patients with acute lymphoblastic leukemia ( ALL ) , and to assess the value of chromosomal examination for the diagnosis and prognosis . METHODS : Fluorescence in situ hybridization ( Q5TCZ1 ) was utilized for detecting the P11274 / P00519 fusion gene and P04637 gene . Median survival time ( MST ) of patients was compared using Log-rank test . RESULTS : Respectively , the MST of patients with white blood cell count ( WBC ) ≤30 × 10(9)/L , normal karyotype , or without a Philadelphia chromosome were significantly greater than those with WBC > 30 × 10(9)/L , abnormal karyotype or Philadelphia chromosome ( P < 0.05 ) . CONCLUSION : WBC , karyotype abnormalities and presence of Philadelphia chromosome are independent factors for the prognosis of ALL in adult patients . The proteomic study of sodium butyrate antiproliferative/cytodifferentiation effects on K562 cells . Employing methods of cell biology and proteome analysis tools , we examined effects of an inhibitor of histone deacetylases , sodium butyrate ( SB ) , on the proliferation/differentiation characteristics of chronic myelogenous leukemia ( CML ) -derived cells K562 . SB suppressed proliferation of K562 cells by inducing cell cycle arrest in P55008 phase , which was followed by their transition to G0 phase ( decrease of Ki-67 antigen-positive cells ) and erythroid differentiation ( increased glycophorin A expression and synthesis of hemoglobins ) . Neither terminal apoptosis ( low counts of TUNEL-positive cells ) nor necrosis ( moderate counts of propidium iodide-positive cells ) occurred . Importantly , SB attenuated protein expression of CML-related chimeric kinase P11274 - P00519 that is responsible for the deregulated proliferation of CML cells . The proteomic analysis ( 2-D electrophoresis combined with MALDI-TOF mass spectrometry and/or Western blotting ) revealed several proteins that were differentially expressed or their mobility was altered due to butyrate treatment , namely , HSP90 , HSP70 , p23 , cyclophilin A ( P62937 ) , prefoldin2 ( Q9UHV9 ) and alpha- , gamma- , epsilon-human globin chains . Perturbation of HSP90 multichaperone complex of which P11274 - P00519 is the client protein is presumably a cause of P11274 - P00519 suppression . Changes in other proteins with chaperonic functions , P62937 and Q9UHV9 , may reflect SB antiproliferative and cytodifferentiation effects . Increased serum levels of neutrophil gelatinase-associated lipocalin in patients with essential thrombocythemia and polycythemia vera . Neutrophil gelatinaase-associated lipocalin ( P80188 ) is a glycoprotein bound with matrix metalloproteinase-9 ( P14780 ) in human neutrophils , and elevated tissue P80188 expression has been documented in different infectious and inflammatory conditions . Recent evidence suggests that P80188 expression is induced in many types of human cancer . Moreover , P80188 is required for P11274 - P00519 -induced tumorigenesis . The aim of the present study was to measure serum levels of P80188 in patients with essential thrombocythemia ( ET ) and polycythemia vera ( PV ) . We also evaluated P80188 levels in patients with ET and PV with and without thrombotic events , to explore a possible correlation of P80188 with platelet and leukocyte activation , and in patients with sepsis . Serum P80188 levels in the study population were significantly higher than in healthy adults and in subjects with sepsis . A correlation between P80188 and the number of white cells and neutrophils was found in patients with PV and ET . P80188 serum levels were not different depending on the presence or not of the O60674 mutation , and a mutant allele dosage effect was not observed for P80188 levels . Patients with PV and ET with thrombosis did not have significantly higher levels of P80188 . We were unable to demonstrate a significant association between serum P80188 levels and CD11b or CD62 expression . In conclusion , our study reports evidence demonstrating that increased levels of P80188 appear to be a characteristic of patients with PV and ET . Differential expression of urokinase-type plasminogen activator and plasminogen activator inhibitor-1 in early and late gestational mouse skin and skin wounds . Early gestation fetal mouse skin heals without scars . P00747 activator inhibitor-1 ( P05121 ) has been associated with postnatal organ fibrosis . We hypothesized that the relative balance between urokinase-type plasminogen activator ( uPA ) and P05121 expression in favor of uPA prevents scarring in early fetal skin wounds , whereas a change in favor of P05121 in late gestation results in wound scarring . To evaluate uPA and P05121 expression , 1-mm skin wounds were made in Q14207 .5 and E18 mice and harvested 24 , 48 , or 96 hours postwounding . DB06692 ( 2 mg/ml ) -coated beads were injected into selected Q14207 .5 wounds . Normal skin and skin wounds were evaluated for uPA , P05121 , and collagen expression . We showed that in normal skin uPA level is higher in Q14207 .5 than in E18 mice , while P05121 is lower in Q14207 .5 than in E18 mice . After wounding , Q14207 .5 wounds show a moderate increase in uPA and a minimal increase in P05121 . E18 wounds show a transient increase in uPA but a significant , sustained increase in P05121 . Addition of aprotinin to Q14207 .5 wounds causes an increase in collagen deposition . We conclude that the differential expression of uPA and P05121 in the skin of early vs. late gestation mice may contribute to the degree of scar formation seen after cutaneous injury . Effect of the hemoregulatory peptide (pEEDCK)2 (pyroGlu- DB00142 - DB00128 - DB00151 -Lys)2 and MIP-1alpha is reduced in bone marrow cultures from patients with chronic myeloid leukemia ( CML ) . The granulocyte-derived hemoregulatory peptide pyroGlu- DB00142 - DB00128 - DB00151 -Lys = pEEDCK is known to keep hematopoietic cells quiescent . When oxidized to its dimeric form (pEEDCK)2 , it activates growth of hematopoietic progenitors in association with stroma-derived cytokines . (pEEDCK)2 has a DB00151 - DB00151 motif which is also a typical feature of the macrophage inflammatory protein ( MIP-1alpha ) . The present study was designed to analyze differences between the response of normal and leukemic progenitor cells to (pEEDCK)2 or MIP-1alpha . When long-term bone marrow cultures ( LTBMCs ) were incubated with (pEEDCK)2 or MIP-1alpha and/or cytokines , the stimulatory effect on colony-forming units-granulocyte/erythroid/macrophage/megakaryocyte of LTBMC from chronic myeloid leukemia ( CML ) patients was less than 50 % compared to LTBMC from healthy humans . No difference in oncogene expression could be observed in LTBMC from CML patients regarding reduction of Philadelphia chromosome-associated transcription of the P11274 - P00519 gene . With respect to the expression of growth and differentiation-associated genes ( Galpha16 , P09917 , phospholipaseA2 , c-kit , and P28906 ) , which were analyzed from LTBMC by semiquantitative reverse transcriptase-polymerase chain reaction , the same transcription rate was observed in CML patients and healthy donors . However , two isoforms of a key enzyme of oxidative metabolism , carnitine palmitoyltransferase ( P50416 and Q92523 ) , showed 50-fold higher expression rates in LTBMC cells of healthy donors compared to CML patients . It is known that a decrease in oxidative metabolism is associated with an increase in redox equivalents in malignancy . This might result in a reduction of disulphide bonds in (pEEDCK)2 or MIP-1alpha , thus inducing a downregulation of these factors in bone marrow from CML patients . Natriuretic and renoprotective effect of chronic oral neutral endopeptidase inhibition in acute renal failure . P08473 ( NEP : EC 3.4.24.11 ) is involved in the degradation of peptides such as atrial natriuretic peptide , angiotensin II ( AngII ) , and endothelin-1 ( ET-1 ) . In this study we propose that NEP inhibition provides protection in glycerol-induced acute renal failure ( Q8N726 ) . Renal vascular responses were evaluated in Q8N726 rats where Q8N726 was induced by injecting 50 % glycerol in candoxatril , a NEP inhibitor ( 30 mg/kg , orally ; for 3 weeks ) pretreated rats . AngII and U46619 ( a TxA2 mimetic ) vasoconstriction was increased ( 2- to 4-fold ) in Q8N726 while ET-1 vasoconstriction was surprisingly reduced ( 23+/-3 % ; p < 0.05 ) . In Q8N726 , candoxatril paradoxically enhanced ET-1 response ( 60+/-20 % ; p < 0.05 ) but reduced AngII vasoconstriction ( 51+/-11 % ; p < 0.05 ) without affecting U46619 response . However , candoxatril treatment was without effect on plasma ET-1 and TxB2 levels in Q8N726 . DB00616 reduced plasma AngII by 34+/-4 % ( p < 0.05 ) in Q8N726 which was approximately 3.5-fold higher compared to control . DB00616 doubled the nitrite excretion in control but was without effect on proteinuria or nitrite excretion in Q8N726 . DB00616 enhanced Na+ and creatinine excretion in Q8N726 by 73+/-9 % and 33+/-2 % , respectively . These results suggest that NEP inhibition may confer protection in glycerol-induced Q8N726 by stimulating renal function but without a consistent effect on renal production and renal vascular responses to endogenous vasoconstrictors . The lipoxygenase-cyclooxygenase inhibitor licofelone prevents thromboxane A2-mediated cardiovascular derangement triggered by the inflammatory peptide fMLP in the rabbit . DB04725 is an analogue of arachidonic acid that inhibits P09917 ( P28300 ) , cyclooxygenase ( P36551 ) -1 and P35354 . We investigated the effects of licofelone on cardiovascular derangements and production of thromboxane (Tx)A(2) induced by the inflammatory agonist n-formyl-methionyl-leucyl-phenylalanine ( fMLP ) in the rabbit , in comparison with those of aspirin or rofecoxib , inhibitors of P23219 and P35354 , respectively . In control rabbits , injection of fMLP ( 30 nmol/kg ) in the jugular vein evokes ischemic electrocardiographic ( ECG ) changes in the first 1-5 min , i.e. a profound depression of the ST segment and inversion of the T wave . Simultaneously , fMLP induces bradycardia and hypotension and increases TxB(2) blood levels . All changes are transient . DB04725 ( 60 mg/kg/5 days , p.os ) prevented fMLP-induced ECG ischemic changes in all treated animals , reverted bradycardia and hypotension , and significantly reduced TxB(2) . DB00945 ( 10 mg/kg/5 days , p.os ) prevented ischemic ECG alterations in 2 out of 5 treated animals and did not modify either bradycardia or hypotension . One rabbit died two min after fMLP . In 2 rabbits , aspirin reduced TxB(2) levels by more than 80 % respect to mean control values ; the remaining two rabbits produced an amount of TxB(2) similar to controls . These two rabbits also showed ischemic ECG changes . DB00533 ( 10 mg/kg/5 days , p.os ) did not prevent fMLP-induced ischemic ECG alteration , bradycardia and hypotension , and did not significantly modify the increase of TxB(2) . These results indicate that the capacity of licofelone to efficiently suppress TxA(2) production , is responsible for the protection from the cardiovascular derangement triggered by an inflammatory stimulus . Regulation of interleukin-6 promoter activation in gastric epithelial cells infected with Helicobacter pylori . The regulation of Helicobacter pylori induced interleukin ( IL ) -6 in the gastric epithelium remains unclear . Primary gastric epithelial cells and MKN28 cells were cocultured with H. pylori and its isogenic cag pathogenicity island ( P05121 ) mutant and/or oipA mutants . H. pylori infection-induced P05231 mRNA expression and P05231 protein production , which was further enhanced by the cag P05121 and OipA . Luciferase reporter gene assays and electrophoretic mobility shift assays showed that full P05231 transcription required binding sites for nuclear factor-kappaB ( NF-kappaB ) , DB02527 response element ( CRE ) , CCAAT/enhancer binding protein ( C/EBP ) , and activator protein ( AP ) -1 . The cag P05121 and OipA were involved in binding to NF-kappaB , AP-1 , CRE , and C/EBP sites . The cag P05121 activated the extracellular signal-regulated kinase ( P29323 ) and Jun N-terminal kinase ( JNK ) pathways ; OipA activated the p38 pathway . Transfection of dominant negative G-protein confirmed roles for Raf , Rac1 , and RhoA in P05231 induction . Overall , the cag P05121 -related P05231 signal transduction pathway involved the Ras/Raf/ Q02750 /2/ P29323 /AP-1/CRE pathway and the JNK/AP-1/CRE pathway ; the OipA-related pathway is p38/AP-1/CRE and both the cag P05121 and OipA appear to be involved in the RhoA/Rac1/NF-kappaB pathway . Combination of different pathways by the cag P05121 and OipA will lead to the maximum P05231 induction . O00206 -dependent induction of vascular adhesion molecule-1 in rheumatoid arthritis synovial fibroblasts : Roles of cytosolic phospholipase A(2)alpha/cyclooxygenase-2 . Lipopolysaccharide ( LPS ) / O00206 ( O00206 ) -mediated signaling pathways have caught the attention of strategies designed for rheumatoid arthritis ( RA ) . In this study , we identified that cPLA(2)alpha acted as a modulator of LPS-induced P19320 expression and THP-1 ( human acute monocytic leukemia cell line ) adherence . Treatment of RA synovial fibroblasts ( RASFs ) with LPS , a O00206 agonist , promoted the P19320 expression and THP-1 adherence which were decreased by pretreatment with a selective cytosolic phospholipase A(2) ( cPLA(2) ) inhibitor ( AACOCF(3) ) , implying the involvement of cPLA(2)alpha in these responses . This notion was further confirmed by knockdown of cPLA(2)alpha expression by transfection with cPLA(2)alpha small interfering RNA ( siRNA ) leading to a decrease in P19320 expression and THP-1 adherence induced by LPS . Subsequently , the LPS-stimulated cPLA(2)alpha phosphorylation was attenuated by pretreatment with a Q02750 /2 inhibitor ( U0126 ) , suggesting that LPS-stimulated cPLA(2)alpha phosphorylation and activity are mediated through an P29323 -dependent mechanism . Moreover , P35354 -derived PGE(2) production appeared to involve in LPS-induced P19320 expression which was attenuated by pretreatment with selective P35354 inhibitors ( NS-398 and celecoxib ) , transfection with P35354 siRNA , or PGE(2) receptor antagonists . In addition , pretreatment with ecosapentaenoic acid ( EPA ) , a substrate competitor of arachidonic acid ( AA ) , also blocked LPS-induced P19320 mRNA and protein expression , and THP-1 adherence . Collectively , these results suggest that LPS-induced P19320 expression and adhesion of THP-1 cells are mediated through the O00206 / P29323 /cPLA(2)alpha phosphorylation and P35354 expression/PGE(2) synthesis in RASFs . Bcr/Abl interferes with the Fanconi anemia/BRCA pathway : implications in the chromosomal instability of chronic myeloid leukemia cells . Chronic myeloid leukemia ( CML ) is a malignant clonal disorder of the hematopoietic system caused by the expression of the P11274 / P00519 fusion oncogene . Although it is well known that CML cells are genetically unstable , the mechanisms accounting for this genomic instability are still poorly understood . Because the Fanconi anemia ( FA ) pathway is believed to control several mechanisms of DNA repair , we investigated whether this pathway was disrupted in CML cells . Our data show that CML cells have a defective capacity to generate Q9BXW9 nuclear foci , either in dividing cells or after DNA damage . Similarly , human cord blood P28906 (+) cells transduced with P11274 / P00519 retroviral vectors showed impaired Q9BXW9 foci formation , whereas Q9BXW9 monoubiquitination in these cells was unaffected . Soon after the transduction of P28906 (+) cells with P11274 / P00519 retroviral vectors a high proportion of cells with supernumerary centrosomes was observed . Similarly , P11274 / P00519 induced a high proportion of chromosomal abnormalities , while mediated a cell survival advantage after exposure to DNA cross-linking agents . Significantly , both the impaired formation of Q9BXW9 nuclear foci , and also the predisposition of P11274 / P00519 cells to develop centrosomal and chromosomal aberrations were reverted by the ectopic expression of P38398 . Taken together , our data show for the first time a disruption of the FA/BRCA pathway in P11274 / P00519 cells , suggesting that this defective pathway should play an important role in the genomic instability of CML by the co-occurrence of centrosomal amplification and DNA repair deficiencies . Purification and characterization of mouse O15528 overproduced by an Escherichia coli system coexpressing molecular chaperonins GroEL/ES . The expression of mouse O15528 in Escherichia coli has been dramatically enhanced by coexpression of GroEL/ES . To reveal the enzymatic properties of O15528 , we measured its hydroxylation activity toward vitamin D3 and DB01436 ( 1alpha(OH)D3 ) in addition to the physiological substrate DB00146 . Surprisingly , O15528 converted vitamin D3 to 1alpha, DB00146 . Both 1alpha-hydroxylation activity toward vitamin D3 , and 25-hydroxylation activity toward 1alpha(OH)D3 were observed . The Km and Vmax values for 25-hydroxylation activity toward 1alpha(OH)D3 were estimated to be 1.7 microM and 0.51 mol/min/mol P450 , respectively , while those for 1alpha-hydroxylation activity toward DB00146 were 0.050 microM and 2.73 mol/min/mol P450 , respectively . Note that the substrate must be fixed in the opposite direction in the substrate-binding pocket of O15528 between 1alpha-hydroxylation and 25-hydroxylation . Based on these results and the fact that human Q02318 and Streptomyces CYP105A1 also convert vitamin D3 to 1alpha, DB00146 , 1alpha-hydroxylation , and 25-hydroxylation of vitamin D3 appear to be closely linked together . Matrix proteinase inhibition by DB05387 , a multifunctional antiangiogenic compound . BACKGROUND : Matrix metalloproteinases ( MMPs ) play an important role in tissue remodeling under normal physiological and pathological conditions and are thus attractive targets for both diagnostic and therapeutic purposes . Here , we examined the effect of DB05387 , an orally bioavailable standardized extract made of cartilage that shows significant antiangiogenic and antimetastatic properties in vivo , on the activity of various members of the MMP family . MATERIALS AND METHODS : The effect of DB05387 on the activity of MMPs was assessed by fluorimetric assays and by substrate gel zymography . RESULTS : DB05387 markedly inhibits the gelatinolytic activity of P08253 and to a lesser extent those of P03956 , P09237 , P14780 and P45452 . DB05387 also inhibited the elastinolytic activities of P08253 and P14780 as well as P39900 ( metalloelastase ) , porcine pancreatic elastase ( PPE ) , and human leukocyte elastase ( P08246 ) . Western blot analysis revealed the presence within DB05387 of immunoreactive P01033 -like proteins , suggesting that these proteins may be at least partly responsible for the observed MMP inhibition . CONCLUSIONS : Taken together , these results demonstrate that DB05387 contains P01033 -like proteins that could be responsible for the specific inhibition of MMPs . Given the recent studies suggesting the presence within this compound of specific inhibitor(s) of endothelial cell proliferation , DB05387 appears as a pleotropic agent able to interfere with several biochemical steps leading to angiogenesis and to other physiopathological conditions . Since DB05387 is currently under Phase III clinical investigations , these findings are also of considerable importance for our understanding of its anticancer properties . Interactions Between Ataxia Telangiectasia Mutated Kinase Inhibition , Poly(ADP-ribose) Polymerase-1 Inhibition and P38398 Status in Breast Cancer Cells . BACKGROUND : Cells harboring P38398 / P51587 mutations are hypersensitive to inhibition of poly(ADP-ribose) polymerase-1 ( P09874 ) . We recently showed that interference with P09874 activity by DB02690 is strongly cytotoxic for P38398 -positive BT-20 cells but not P38398 -deficient SKBr-3 cells . These unexpected observations prompted speculation that other P09874 inhibitor(s) may be more cytotoxic towards SKBr-3 cells . In addition , interference with the DNA damage signaling pathway via ( for instance ) Q13315 ( Q13315 ) kinase inhibition may induce synthetic lethality in DNA repair-deficient breast cancer cells and pharmacological interference with Q13315 activity may sensitize breast cancer cells to P09874 inactivation . METHODS : We determined drug cytotoxicity in human MCF-7 and SKBr-3 breast cancer cells using the CellTiterGLO Luminescent cell viability assay and a Tecan multi-label , multitask plate counter to measure generated luminescence . Changes in cell cycle progression were monitored by flow cytometric measurement of DNA content in cells stained with propidium iodide . RESULTS : Unlike DB02690 , AZD2461 , a new P09874 inhibitor , markedly reduced the numbers of living MCF-7 and SKBr-3 cells . Q13315 kinase inhibition ( CP466722 ) was also cytotoxic for both MCF-7 and SKBr-3 cells . Furthermore , AZD2461 enhanced the cytotoxicity of CP466722 in both cell lines by inducing apoptosis , and concurrent inhibition of Q13315 and P09874 reduced cell proliferation more strongly than either single treatment . CONCLUSIONS : Our data show that inhibition of P09874 by AZD2461 is synthetically lethal for DB02690 -resistant MCF-7 and SKBr-3 breast cancer cells . They also indicate that DNA damage signaling is essential for survival of both SKBr-3 and MCF-7 cells , especially after inactivation of P09874 . Expression of Th2-skewed pathology mediators in monocyte-derived type 2 of dendritic cells ( DC2 ) . The information conveyed from dendritic cells ( DCs ) to naïve P01730 (+) T cells has crucial influence on their differentiation toward effector T cells . In an effort to identify DC-derived molecules directly contributing to T cell differentiation , we searched for molecules distinctively expressed between two DC subtypes , which were differentiated from peripheral monocytes by cultivation with GM- P04141 ( for Q9NPG8 ) or P08700 ( for DC2 ) in the presence of P05112 and had the ability to induce naïve T cells to differentiate into Th1 or Th2 cells , respectively . As the first step to address this issue , we subtracted Q9NPG8 transcripts from those of DC2 and compiled the gene profile dominantly expressed in DC2 , whose products are known to reside in other than the nucleus . Intriguingly , many of them were molecules involved in Th2-skewed disease pathologies , such as P02751 , P38570 , Q14956 , Q03405 , P25089 , Q8NHJ6 , P05121 , P16050 , P24557 , P19878 , P10147 , P18510 , P09486 , and Q9NY15 , suggesting that DCs function not only as antigen presenting cells but also as producers of Th2 pathology specific milieus leading to disease deteriorations . We also found that expressions of Q02318 , O14495 , Q8WXG1 , and O15438 were up-regulated in DC2 , implying their significant function in Th2-deviated states . The identification of differentially expressed genes between DC subtypes provides new insights into their functions and our comparative gene expression profile will be highly useful for the identification of DC-derived key molecules for T cell differentiation .

MH_train_1380

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MH_train_1380

interacts_with DB00477?

multiple_choice

Raddeanin A , a triterpenoid saponin isolated from Anemone raddeana , suppresses the angiogenesis and growth of human colorectal tumor by inhibiting P35968 signaling . Raddeanin A ( RA ) is an active triterpenoid saponin from a traditional Chinese medicinal herb , Anemone raddeana Regel . It was previously reported that RA possessed attractive antitumor activity through inhibiting proliferation and inducing apoptosis of multiple cancer cells . However , whether RA can inhibit angiogenesis , an essential step in cancer development , remains unknown . In this study , we found that RA could significantly inhibit human umbilical vein endothelial cell ( HUVEC ) proliferation , motility , migration , and tube formation . RA also dramatically reduced angiogenesis in chick embryo chorioallantoic membrane ( P62158 ) , restrained the trunk angiogenesis in zebrafish , and suppressed angiogenesis and growth of human HCT-15 colorectal cancer xenograft in mice . Western blot assay showed that RA suppressed P15692 -induced phosphorylation of P35968 and its downstream protein kinases including PLCγ1 , O60674 , Q05397 , Src , and Akt . Molecular docking simulation indicated that RA formed hydrogen bonds and hydrophobic interactions within the DB00171 binding pocket of P35968 kinase domain . Our study firstly provides the evidence that RA has high antiangiogenic potency and explores its molecular basis , demonstrating that RA is a potential agent or lead candidate for antiangiogenic cancer therapy . Proliferation of endothelial cell on polytetrafluoroethylene vascular graft materials carried P15692 gene plasmid . OBJECTIVE : To investigate whether vascular endothelial growth factor ( P15692 ) gene plasmid carried by polytetrafluoroethylene ( PTFE ) vascular graft materials could transfect endothelial cells ( ECs ) and promote their growth . METHODS : PTFE vascular graft materials carried with pCDI-hVEGF(121) , pCDI or pEGFP were incubated in DB03754 -buffer solution and the values of optical density of 260 nm at different time were plotted , then the DNA controlled release curve was made . ECs derived from human umbilical vein were seeded on the pCDI-hVEGF(121)/pCDI/pEGFP-PTFE materials or tissue culture plates , ECs numbers were counted and P15692 protein concentrations at different time were measured by enzyme-linked immunoadsorbent assay method . Green fluorescent protein ( GFP ) expression in ECs on pEGFP-PTFE materials was examined with fluorescence microscopy . RESULTS : The controlled release curve showed that the gene released from PTFE materials was rapid within 8 h , then slowed down and that the gene released continuously even after 72 h . At 24 , 72 and 120 h , ECs number and proliferation rate of pCDI-hVEGF(121)-PTFE materials were higher than those of pCDI or pEGFP-PTFE materials ( P < 0.05 ) . P15692 protein concentration of pCDI-hVEGF(121)-PTFE materials was higher than that of pCDI or pEGFP-PTFE materials at 6 , 24 , 72 and 120 h ( P < 0.01 ) . GFP expression in ECs on the pEGFP-PTFE materials could be detected by fluorescence microscopy . CONCLUSION : PTFE graft can be used as a carrier of P15692 gene plasmid , P15692 gene carried by PTFE can transfect ECs and promote ECs growth . Proangiogenesis action of the thyroid hormone analog 3,5-diiodothyropropionic acid ( DITPA ) is initiated at the cell surface and is integrin mediated . We have recently described the proangiogenesis effects of thyroid hormone in the chick chorioallantoic membrane ( P62158 ) model . Generation of new blood vessels from existing vessels was promoted 2- to 3-fold by either T(4) or T(3) at 10(-8)-10(-7) M total hormone concentrations . In the present studies , nanomolar concentrations of 3,5-diiodothyropropionic acid ( DITPA ) , a thyroid hormone analog with inotropic but not chronotropic properties , exhibited potent proangiogenic activity that was comparable to that obtained with T(3) and T(4) in both the P62158 model and in an in vitro three-dimensional human microvascular endothelial sprouting assay . The proangiogenesis effect of DITPA was inhibited by tetraiodothyroacetic acid , a thyroid hormone analog that competes with T(4) and T(3) for a novel cell surface hormone receptor site on integrin alphavbeta3 . The thyroid hormone analogs DITPA , T(4) , and T(4)-agarose , as well as basic fibroblast growth factor ( b-FGF ) and vascular endothelial cell growth factor , demonstrated comparable proangiogenic effects in the P62158 model and in the three-dimensional human microvascular endothelial sprouting model . The proangiogenesis effect of either DITPA or b-FGF was blocked by PD 98059 , an inhibitor of the P27361 /2 signal transduction cascade . Additionally , a specific integrin alphavbeta3 small molecule antagonist , XT199 , effectively inhibited the proangiogenesis effect of DITPA and b-FGF . Thus , the proangiogenesis actions of thyroid hormone and its analog DITPA are initiated at the plasma membrane , apparently at integrin alphavbeta3 , and are MAPK dependent . Implantation of P15692 transfected preadipocytes improves vascularization of fibrin implants on the cylinder chorioallantoic membrane ( P62158 ) model . The successful substitution or augmentation of soft tissues by implantation of three dimensional cell constructs , consisting of human preadipocytes and fibrin glue as a carrier matrix , requires a rapid and homogeneous vascularization of the whole implant in order to provide a sufficient blood supply of centrally situated cells . Previous investigations have shown that under in vivo conditions primary human preadipocytes induce vascularization of fibrin matrices by secretion of several growth factors , such as P15692 and P09038 . The current study investigates whether vascularization of implants can be improved by transplantation of preadipocytes following transfection with a P15692 -vector . Transfection was performed by electroporation with an pCMX-GFP and pCMX-VEGF165 vector . Transfection efficiency ( GFP expression ) and P15692 expression were determined in vitro by FACS analysis and P15692 immunoassay , respectively . In vivo investigations to determine the vascularization of the implants were performed on the cylinder chorioallantoic membrane ( P62158 ) . Four million P15692 transfected cells were transferred within a fibrin matrix onto the P62158 on the 7(th) day of incubation and after 8 days the vascularization of the implant was histologically examined and evaluated by means of a computer-assisted image analysis program . Transfection of preadipocytes with the GFP vector by electroporation yielded transfection efficiencies between 12 % and 41 % of surviving cells . Results of the P15692 immunoassay demonstrated that P15692 expression was significantly higher following transfection . Investigations on the P62158 outlined a significantly higher rate of vascularization in the transfected vs. control population . Our investigations demonstrate that primary human preadipocytes can be successfully transfected by electroporation with a P15692 vector . The enhanced P15692 expression on transfected cells results in an increase of vascularization of the cell constructs on the P62158 . P43490 / P43490 /visfatin and cancer . P43490 / P43490 /visfatin is the rate-limiting enzyme that catalyzes the first step in NAD biosynthesis from nicotinamide and regulates growth , apoptosis and angiogenesis of mammalian cells . This enzyme was originally cloned as a putative cytokine shown to enhance the B cell precursor maturation in the presence of P13232 and stem cell factor . A number of cancers have increased expression of P43490 / P43490 /visfatin , which regulates a variety of different signaling pathways such as PI3K/Akt , P27361 /2 and P40763 . FK866/APO866 and CHS828/ DB05217 are two known inhibitors of P43490 / P43490 /visfatin and have been evaluated as anticancer agents in the clinic . This review will focus on its role in carcinogenesis and cancer progression and its inhibitors as therapeutic target for cancer treatment . SAR and in vivo evaluation of 4-aryl-2-aminoalkylpyrimidines as potent and selective O60674 ( O60674 ) inhibitors . We report the discovery of a series of 4-aryl-2-aminoalkylpyrimidine derivatives as potent and selective O60674 inhibitors . High throughput screening of our in-house compound library led to the identification of hit 1 , from which optimization resulted in the discovery of highly potent and selective O60674 inhibitors . Advanced lead 10d demonstrated a significant dose-dependent pharmacodynamic and antitumor effect in a mouse xenograft model . Based upon the desirable profile of 10d ( DB05243 ) it was advanced into clinical trials . Role of methionine adenosyltransferase α2 and β phosphorylation and stabilization in human hepatic stellate cell trans-differentiation . Myofibroblastic trans-differentiation of hepatic stellate cells ( HSCs ) is an essential event in the development of liver fibrogenesis . These changes involve modulation of key regulators of the genome and the proteome . DB00134 adenosyltransferases ( P24752 ) catalyze the biosynthesis of the methyl donor , S-adenosylmethionine ( DB00118 ) from methionine . We have previously shown that two P24752 genes , P31153 and Q9NZL9 ( encoding MATα2 and MATβ proteins respectively ) , are required for P19526 activation and loss of P31153 transcriptional control favors its up-regulation during trans-differentiation . Hence P24752 genes are intrinsically linked to the P19526 machinery during activation . In the current study , we have identified for the first time , post-translational modifications in the MATα2 and MATβ proteins that stabilize them and favor human P19526 trans-differentiation . Culture-activation of human HSCs induced the MATα2 and MATβ proteins . Using mass spectrometry , we identified phosphorylation sites in MATα2 and MATβ predicted to be phosphorylated by mitogen-activated protein kinase ( MAPK ) family members ( P27361 /2 , V-Raf Murine Sarcoma Viral Oncogene Homolog B1 [ B-Raf ] , MEK ) . Phosphorylation of both proteins was enhanced during P19526 activation . Blocking MEK activation lowered the phosphorylation and stability of P24752 proteins without influencing their mRNA levels . Silencing P27361 /2 or B-Raf lowered the phosphorylation and stability of MATβ but not MATα2 . Reversal of the activated human P19526 cell line , LX2 to quiescence lowered phosphorylation and destabilized P24752 proteins . Mutagenesis of MATα2 and MATβ phospho-sites destabilized them and prevented P19526 trans-differentiation . The data reveal that phosphorylation of P24752 proteins during P19526 activation stabilizes them thereby positively regulating trans-differentiation . Phosphodiesterase type 4 isozymes expression in human brain examined by in situ hybridization histochemistry and[3H]rolipram binding autoradiography . Comparison with monkey and rat brain . We have examined the distribution of four different cyclic AMP-specific phosphodiesterase isozyme ( P27815 , Q07343 , Q08493 and Q08499 ) mRNAs in the brain of different species by in situ hybridization histochemistry and by autoradiography with [3H]rolipram . We have compared the localization of each isozyme in human brain with that in rat and monkey brain . We have found that the four DB05876 isoforms display a differential expression pattern at both regional and cellular level in the three species . P27815 , Q07343 and Q08499 are widely distributed in human brain , with the two latter appearing more abundant . In contrast , Q08493 in human brain , presents a more restricted distribution , limited to cortex , some thalamic nuclei and cerebellum . This is at variance with the distribution of Q08493 in rat brain , where it is found exclusively in olfactory bulb . In monkey brain , the highest expression for this isoform is found in the claustrum , and at lower levels in cortical areas and cerebellum . Q07343 presented a broad distribution , being expressed in both neuronal and non neuronal cell populations . In general , the distribution of binding sites visualized with [3H]rolipram correlated well with the expression of each DB05876 isozyme . Expression of human all-trans-retinoic acid receptor beta and its ligand-binding domain in Escherichia coli . DB00755 , one of the hormonally active derivatives of vitamin A , occurs physiologically in plasma at a concentration below 10 nmol/l . The methods currently used for its quantification are based on HPLC , need about 1 ml of serum , are relatively laborious and thus not well suited for mass analysis . The affinity and specificity of retinoic acid receptors for all-trans-retinoic acid encouraged us to express both the entire human retinoic acid receptor beta ( P10826 ) and two versions of its retinoic acid-binding domain in Escherichia coli in the hope that these recombinant proteins might be used as binders in a ligand-binding assay for all-trans-retinoic acid . The recombinant receptors , the whole receptor [ P10826 -( Q93033 -Q448) ] , corresponding to domains A-F , and the ligand-binding domain [ P10826 -(E149-Q448) ] , corresponding to domains D-F , were expressed in the vector pET 3d/BL21 ( DE3 ) as inclusion bodies , solubilized with guanidinium chloride , renatured and purified by ion-exchange chromatography . P10826 -(P193-Q448) , corresponding to domains E-F , was expressed in the vector pET 3d/BL21(DE3)pLysS , and purified by reversed-phase chromatography . Under non-denaturing conditions , the expressed whole receptor [ P10826 -( Q93033 -Q448) ] and the D-F construct ( P10826 -(E149-Q448) ] behaved chromatographically as monomeric proteins whereas the E-F construct [ P10826 -(P193-Q448) ] had a strong tendency to aggregate . P10826 -( Q93033 -Q448) and P10826 -(E149-Q448) had similar Kd values for all-trans-retinoic acid ( 1.4 and 0.6 nmol/l respectively ) whereas P10826 -(P193-Q448) bound all-trans-retinoic acid less avidly ( Kd 9.6 nmol/l ) . DB00523 bound to P10826 -(E149-Q448) and P10826 -( Q93033 -Q448) as avidly as all-trans-retinoic acid . Competition experiments showed weak or no binding of 4-oxo-all-trans-retinoic acid , 4-oxo-13-cis-retinoic acid , 13-cis-retinoic acid , acitretin and retinol by P10826 -(E149-Q448) . Brucine suppresses colon cancer cells growth via mediating P35968 signalling pathway . Angiogenesis plays an important role in colon cancer development . This study aimed to demonstrate the effect of brucine on tumour angiogenesis and its mechanism of action . The anti-angiogenic effect was evaluated on the chicken chorioallantoic membrane ( P62158 ) model and tube formation . The mechanism was demonstrated through detecting mRNA and protein expressions of P35968 ( P35968 ) , PKCα , PLCγ and Raf1 by reverse transcription-polymerase chain reaction ( RT-PCR ) and Western blot ( WB ) , as well as expressions of P15692 and PKCβ and P42345 by ELISA and WB . The results showed that brucine significantly reduced angiogenesis of P62158 and tube formation , inhibited the P15692 secretion and P42345 expression in LoVo cell and down-regulated the mRNA and phosphorylation protein expressions of P35968 , PKCα , PLCγ and Raf1 . In addition , the effects of brucine on P35968 kinase activity , viability of LoVo cell and gene knockdown cell were detected with the Lance™ assay , WST-1 assay and instantaneous siRNA . Compared to that of normal LoVo cells , the inhibition on proliferation of knockdown cells by brucine decreased significantly . These results suggest that brucine could inhibit angiogenesis and be a useful therapeutic candidate for colon cancer intervention . Characterization of DB02527 degradation by phosphodiesterases in the accessory olfactory system . To characterize the potential role of DB02527 in pheromone transduction , we have examined the occurrence of cyclic nucleotide phosphodiesterases ( PDEs ) in the mouse vomeronasal organ ( VNO ) . We show that the DB02527 -specific isoforms P27815 and Q08499 are found preferentially in the apical and basal layers , respectively , of the VNO neuroepithelium and in the rostral ( P27815 ) and caudal ( Q08499 ) portions of the accessory olfactory bulb glomerular layer . Assays for DB02527 hydrolysis showed that PDE activity in VNO homogenates was about half that measured in the cerebral cortex and olfactory epithelium , and the proportion of total activity inhibited by rolipram , a DB05876 -specific inhibitor , was approximately 40 % . Activity in the VNO was enhanced 60 % by Ca(2+) and calmodulin ( P62158 ) , implicating the presence of Ca(2+)/ P62158 -dependent PDE1 . Zaprinast , which is known to inhibit Q14123 isoforms , completely suppressed Ca(2+)/ P62158 -stimulated activity and , together , zaprinast and rolipram inhibited DB02527 hydrolysis by approximately 70 % . Our results suggest that PDE1 and DB05876 isoforms are the primary source of DB02527 degradation in the VNO . Oral leucine supplementation is sensed by the brain but neither reduces food intake nor induces an anorectic pattern of gene expression in the hypothalamus . DB00149 activates the intracellular mammalian target of the rapamycin ( P42345 ) pathway , and hypothalamic P42345 signaling regulates food intake . Although central infusion of leucine reduces food intake , it is still uncertain whether oral leucine supplementation is able to affect the hypothalamic circuits that control energy balance . We observed increased phosphorylation of p70s6k in the mouse hypothalamus after an acute oral gavage of leucine . We then assessed whether acute oral gavage of leucine induces the activation of neurons in several hypothalamic nuclei and in the brainstem . DB00149 did not induce the expression of Fos in hypothalamic nuclei , but it increased the number of Fos-immunoreactive neurons in the area postrema . In addition , oral gavage of leucine acutely increased the 24 h food intake of mice . Nonetheless , chronic leucine supplementation in the drinking water did not change the food intake and the weight gain of ob/ob mice and of wild-type mice consuming a low- or a high-fat diet . We assessed the hypothalamic gene expression and observed that leucine supplementation increased the expression of enzymes ( P54687 , O15382 and O14874 ) that metabolize branched-chain amino acids . Despite these effects , leucine supplementation did not induce an anorectic pattern of gene expression in the hypothalamus . In conclusion , our data show that the brain is able to sense oral leucine intake . However , the food intake is not modified by chronic oral leucine supplementation . These results question the possible efficacy of leucine supplementation as an appetite suppressant to treat obesity . Candidate genes for cannabis use disorders : findings , challenges and directions . AIM : Twin studies have shown that cannabis use disorders ( abuse/dependence ) are highly heritable . This review aims to : ( i ) review existing linkage studies of cannabis use disorders and ( ii ) review gene association studies , to identify potential candidate genes , including those that have been tested for composite substance use disorders and ( iii ) to highlight challenges in the genomic study of cannabis use disorders . METHODS : Peer-reviewed linkage and candidate gene association studies are reviewed . RESULTS : Four linkage studies are reviewed : results from these have homed in on regions on chromosomes 1 , 3 , 4 , 9 , 14 , 17 and 18 , which harbor candidates of predicted biological relevance , such as monoglyceride lipase ( Q99685 ) on chromosome 3 , but also novel genes , including Q9HBW9 [ epidermal growth factor ( P01133 ) , latrophilin and seven transmembrane domain containing 1 ] on chromosome 1 . Gene association studies are presented for ( a ) genes posited to have specific influences on cannabis use disorders : P21554 , CB2 , FAAH , Q99685 , Q8NER1 and Q9Y2T6 and ( b ) genes from various neurotransmitter systems that are likely to exert a non-specific influence on risk of cannabis use disorders , e.g. P47869 , P14416 and P35372 . CONCLUSIONS : There are challenges associated with ( i ) understanding biological complexity underlying cannabis use disorders ( including the need to study gene-gene and gene-environment interactions ) , ( ii ) using diagnostic versus quantitative phenotypes , ( iii ) delineating which stage of cannabis involvement ( e.g. use versus misuse ) genes influence and ( iv ) problems of sample ascertainment . P16473 antibodies associated with post-operative relapse of thyrotoxicosis in a pregnancy complicated by neonatal thyrotoxicosis . This case describes the clinical , biochemical and immunological features associated with relapse of thyrotoxicosis during pregnancy in a patient who had recently undergone a subtotal thyroidectomy for Graves ' disease . The baby , shortly after birth , showed clinical and biochemical features of thyrotoxicosis which responded to carbimazole therapy . P16473 antibodies and thyroid stimulating antibodies were present in the blood of the mother and baby . The clinical course of the neonatal thyrotoxicosis correlated with the DB00024 receptor antibody levels . P35372 phosphorylation , desensitization , and ligand efficacy . Mu opioid receptors are subject to phosphorylation and desensitization through actions of at least two distinct biochemical pathways : agonist-dependent mu receptor phosphorylation and desensitization induced by a biochemically distinct second pathway dependent on protein kinase C activation ( 1 ) . To better understand the nature of the agonist-induced mu receptor phosphorylation events , we have investigated the effects of a variety of opiate ligands of varying potencies and intrinsic activities on mu receptor phosphorylation and desensitization . Exposure to the potent full agonists sufentanil , dihydroetorphine , etorphine , etonitazine , and [ D-Ala2 , MePhe4 , Glyol5 ] enkephalin ( DAMGO ) led to strong receptor phosphorylation , while methadone , l-alpha-acetylmethadone ( DB01227 ) , morphine , meperidine , DADL , beta-endorphin(1-31) , enkephalins , and dynorphin A(1-17) produced intermediate effects . The partial agonist buprenorphine minimally enhanced receptor phosphorylation while antagonists failed to alter phosphorylation . DB00921 and full antagonists each antagonized the enhanced mu receptor phosphorylation induced by morphine or DAMGO . The rank order of opiate ligand efficacies in producing mu receptor-mediated functional desensitization generally paralleled their rank order of efficacies in producing receptor phosphorylation . Interestingly , the desensitization and phosphorylation mediated by methadone and DB01227 were disproportionate to their efficacies in two distinct test systems . This generally good fit between the efficacies of opiates in mu receptor activation , phosphorylation , and desensitization supports the idea that activated receptor/agonist/G-protein complexes and/or receptor conformational changes induced by agonists are required for agonist-induced mu receptor phosphorylation . Data for methadone and DB01227 suggest possible contribution from their enhanced desensitizing abilities to their therapeutic efficacies . An P01133 receptor inhibitor induces P10826 expression in breast and ovarian cancer cells . Inhibition of the epidermal growth factor ( P01133 ) -receptor ( P00533 ) has become a promising anticancer treatment strategy . In addition , application of retinoids yields encouraging results for cancer prevention and therapy . Many tumors express no or low amounts of retinoic acid receptor-beta2 ( RAR-beta2 ) due to epigenetic silencing via DNA hypermethylation . RAR-beta2 is the main mediator of the antiproliferative effect of retinoids . RAR-beta2 re-expression causes reversal of transformation , cell cycle arrest , and restoration of retinoid sensitivity . RAR-beta2 is thus a tumor suppressor . Western blotting , colorimetric in vitro cell proliferation assays , and reverse transcription-polymerase chain reaction demonstrated that the P00533 inhibitor PD153035 not only blocked activation of P00533 and inhibited cell growth , but also stimulated P10826 expression in MDA-MB-468 breast and OVCAR-3 ovarian carcinoma cells . Upregulation of P10826 by PD153035 was confirmed by real-time reverse transcription-polymerase chain reaction . In contrast , expression of other retinoid receptors and of estrogen receptor-alpha was not affected . PD153035-mediated re-induction of P10826 was associated with demethylation of the RAR-beta2 gene promoter P2 as demonstrated by methylation-specific polymerase chain reaction . These novel results on the ErbB/retinoid receptor cross-talk may be useful for designing future anticancer combination regimens . [ Functional characteristics of calcium-sensitive adenylyl cyclase of ciliate Tetrahymena pyriformis ] . DB01373 -sensitive forms of adenylyl cyclase ( AC ) were revealed in most vertebrates and invertebrates and also in some unicellular organisms , in particular ciliates . We have shown for the first time that calcium cations influence the AC activity of ciliate Tetrahymena pyriformis . These cations at the concentrations of 0.2-20 microM stimulated the enzyme activity , and maximum of catalytic effect was observed at 2 microM Ca2+ . DB01373 cations at a concentrations of 100 microM or higher inhibited the AC activity . P62158 antagonists W-5 and W-7 at the concentrations of 20-100 microM inhibited the catalytic effect induced by 5 microM Ca2+ and blocked the effect at higher concentrations of Ca2+ . DB00477 , another calmodulin antagonist , reduced Ca2+-stimulated AC activity only at the concentrations of 200-1000 microM . AC stimulating effects of serotonin , P01133 and DB02527 increased in the presence of 5 microM Ca2+ . AC stimulating effects of P01133 , DB02527 and insulin decreased in the presence of 100 microM Ca2+ , and AC stimulating effect of DB02527 decreased also in the presence of calmodulin antagonists ( 1 mM ) . At the same time , stimulating effect of D-glucose in the presence of Ca2+ and calmodulin antagonists did not change essentially . The data obtained speak in favor of the presence of calcium-sensitive forms of AC in ciliate T. pyriformis which mediate enzyme stimulation by P01133 , DB02527 , insulin , and serotonin . Molecular sampling of the allosteric binding pocket of the DB00024 receptor provides discriminative pharmacophores for antagonist and agonists . The P16473 ( thyrotropin receptor ) is activated endogenously by the large hormone thyrotropin and activated pathologically by auto-antibodies . Both activate and bind at the extracellular domain . Recently , SMLs ( small-molecule ligands ) have been identified , which bind in an allosteric binding pocket within the transmembrane domain . Modelling driven site-directed mutagenesis of amino acids lining this pocket led to the delineation of activation and inactivation sensitive residues . Modified residues showing CAMs ( constitutively activating mutations ) indicate signalling-sensitive positions and mark potential trigger points for agonists . Silencing mutations lead to an impairment of basal activity and mark contact points for antagonists . Mapping these residues on to a structural model of P16473 indicates locations where an SML may switch the receptor to an inactive or active conformation . In the present article , we report the effects of SMLs on these signalling-sensitive amino acids at the P16473 . Surprisingly , the antagonistic effect of SML compound 52 was reversed to an agonistic effect , when tested at the P62158 Y667A . Switching agonism to antagonism and the reverse by changing either SMLs or residues covering the binding pocket provides detailed knowledge about discriminative pharmacophores . It prepares the basis for rational optimization of new high-affinity antagonists to interfere with the pathogenic activation of the P16473 .

MH_train_1381

MH_train_1381

MH_train_1381

interacts_with DB01259?

multiple_choice

Nearly Complete Response of Brain Metastases from P04626 Overexpressing Breast Cancer with DB01259 and DB01101 after Whole Brain Irradiation . DB00072 treatment does not prevent intracranial seeding and is largely ineffective for established central nervous system metastasis in P04626 overexpressing breast cancer patients . Combination therapy of lapatinib and capecitabine may be an effective treatment option for brain metastasis of P04626 -positive breast cancer . We report a patient with breast cancer overexpressing HER-2 where brain metastases were successfully treated with radiation and a combination of lapatinib and capecitabine . [ Gene expression of P08138 , P00533 , P01215 , P01303 in 4 neuroblastoma cell lines ] . Gene expression of nerve growth factor receptor ( P08138 ) , epidermal growth factor receptor( P00533 ) , chromogranin A ( P01215 ) and neuropeptide Y ( P01303 ) in 4 neuroblastoma cell Lines without N-myc amplification was studied by using Northern blot technique . N type cells expressed more P08138 mRNA than S type cell 's and have only little or no P00533 expression . S type cells had stronger expression of P00533 mRNA than that of N type cells accompanying with only less or even no P08138 expression . The results indicated that difference of gene expression of these growth factor receptors might be due to the various directions of tumor cell differentiation . Cells differentiating toward neurons gave more P08138 expression and cells prepared to be differentiating toward other direction might give more P00533 gene expression . Various gene expression of P01215 and P01303 in neuroblastoma cell lines might be due to the presence of different stages of tumor cell differentiation and P01138 only induced differentiation of those neuroblastoma cells ready to be differentiating to neurons afterwards . A PH domain in the Arf P20936 ( P20936 ) Q96P48 binds phosphatidylinositol 3,4,5-trisphosphate and regulates Arf P20936 activity independently of recruitment to the plasma membranes . Q96P48 is a phosphatidylinositol 3,4,5-trisphosphate ( PtdIns(3,4,5)P(3) ) -dependent Arf P20936 ( P20936 ) with five PH domains that regulates endocytic trafficking of the epidermal growth factor receptor ( P00533 ) . Two tandem PH domains are immediately N-terminal of the Arf P20936 domain , and one of these fits the consensus sequence for PtdIns(3,4,5)P(3) binding . Here , we tested the hypothesis that PtdIns(3,4,5)P(3)-dependent recruitment mediated by the first PH domain of Q96P48 regulates the in vivo and in vitro function of Q96P48 . We found that P78364 of Q96P48 specifically bound to PtdIns(3,4,5)P(3) , but with relatively low affinity ( approximately 1.6 microm ) , and the PH domains did not mediate PtdIns(3,4,5)P(3)-dependent recruitment to membranes in cells . However , PtdIns(3,4,5)P(3) binding to the PH domain stimulated P20936 activity and was required for in vivo function of Q96P48 as a regulator of endocytic trafficking of the P00533 . Based on these results , we propose a variation on the model for the function of phosphoinositide-binding PH domains . In our model , Q96P48 is recruited to membranes independently of PtdIns(3,4,5)P(3) , the subsequent production of which triggers enzymatic activity . Dynamic simulations of pathways downstream of P00533 -family , including mutations and treatments : concordance with experimental results . The pathways downstream of ErbB-family proteins are very important in BC , especially when considering treatment with onco-protein inhibitors . We studied and implemented dynamic simulations of four downstream pathways and described the fragment of the signaling network we evaluated as a Molecular Interaction Map . Our simulations , enacted using Ordinary Differential Equations , involved 242 modified species and complexes , 279 reversible reactions and 111 catalytic reactions . Mutations within a single pathway tended to be mutually exclusive ; only inhibitors acting at , or downstream ( not upstream ) , of a given mutation were active . A double alteration along two distinct pathways required the inhibition of both pathways . We started an analysis of sensitivity/robustness of our network , and we systematically introduced several individual fluctuations of total concentrations of independent molecular species . Only very few cases showed significant sensitivity . We transduced the ErbB2 over-expressing BC line , BT474 , with the P01112 ( V12 ) mutant , then treated it with ErbB-family and phosphorylated MEK ( MEKPP ) inhibitors , DB01259 and U0126 , respectively . Experimental and simulation results were highly concordant , showing statistical significance for both pathways and for two respective endpoints , i.e. phosphorylated active forms of P29323 and Akt , p one tailed = .0072 and = .0022 , respectively . Working with a complex 39 basic species signaling network region , this technology facilitates both comprehension and effective , efficient and accurate modeling and data interpretation . Dynamic network simulations we performed proved to be both practical and valuable for a posteriori comprehension of biological networks and signaling , thereby greatly facilitating handling , and thus complete exploitation , of biological data . Regional selective neuronal degeneration after protein phosphatase inhibition in hippocampal slice cultures : evidence for a Q96HU1 kinase-dependent mechanism . The regional selectivity and mechanisms underlying the toxicity of the serine/threonine protein phosphatase inhibitor okadaic acid ( OA ) were investigated in hippocampal slice cultures . Image analysis of propidium iodide-labeled cultures revealed that okadaic acid caused a dose- and time-dependent injury to hippocampal neurons . Pyramidal cells in the P07451 region and granule cells in the dentate gyrus were much more sensitive to okadaic acid than the pyramidal cells in the P00915 region . Electron microscopy revealed ultrastructural changes in the pyramidal cells that were not consistent with an apoptotic process . Treatment with okadaic acid led to a rapid and sustained tyrosine phosphorylation of the mitogen-activated protein kinases P27361 and P28482 ( Q8TCB0 /42(mapk) ) . The phosphorylation was markedly reduced after treatment of the cultures with the microbial alkaloid DB02152 ( a nonselective protein kinase inhibitor ) or the Q96HU1 kinase kinase ( Q02750 /2 ) inhibitor PD98059 . DB02152 and PD98059 also ameliorated the okadaic acid-induced cell death . Inhibitors of protein kinase C , Ca2+/calmodulin-dependent protein kinase II , or tyrosine kinase were ineffective . These results indicate that sustained activation of the Q96HU1 kinase pathway , as seen after e.g. , ischemia , may selectively harm specific subsets of neurons . The susceptibility to Q96HU1 kinase activation of the P07451 pyramidal cells and dentate granule cells may provide insight into the observed relationship between cerebral ischemia and dementia in Alzheimer 's disease . DB01645 stimulates duodenal HCO(3)(-) secretion through PI3K pathway in mice . DB01645 has been proposed as a promising pharmacotherapeutic for cystic fibrosis . We recently found that genistein stimulates murine duodenal HCO(3)(-) secretion through cystic fibrosis transmembrane conductance regulator ( P13569 ) . The aim of the present study was to determine the intracellular signal pathways involved in genistein-stimulated duodenal HCO(3)(-) secretion . Murine duodenal mucosal HCO(3)(-) secretion was examined in vitro in Ussing chambers by the pH-stat technique . The results showed that neither DB02527 -dependent signal pathway inhibitors MDL-12330A and KT-5720 , nor cGMP signal pathway inhibitors NS2028 and KT5823 , nor calcium signal pathway inhibitors verapamil and W-13 , altered genistein-stimulated duodenal HCO(3)(-) secretion . In calcium-free solution , genistein-stimulated duodenal HCO(3)(-) secretion was not altered either . Vanadate , an inhibitor of protein tyrosine phosphatase , only partially inhibited genistein-stimulated duodenal HCO(3)(-) secretion . However , both wortmannin and LY294002 , two structurally and mechanistically distinct phosphatidylinositol 3-kinase ( PI3K ) inhibitors , markedly inhibited genistein-stimulated duodenal HCO(3)(-) secretion . DB01645 increased duodenal mucosal PI3K activity and induced the phosphorylation of Akt , a signaling molecule downstream of PI3K , which was again inhibited by wortmannin . P03372 antagonist , ICI182,780 , also markedly inhibited genistein-stimulated duodenal HCO(3)(-) secretion and genistein-induced PI3K activity increase in duodenal mucosa . These results demonstrate that genistein stimulates duodenal HCO(3)(-) secretion mainly through estrogen receptor and PI3K-dependent pathway . These findings contribute to the understanding of the molecular mechanism of genistein-induced anion secretion and further pharmacotherapeutic development and use of genistein or related substances in the treatment of diseases of epithelial tissues . Coexpression of P49771 and GM- P04141 genes modulates immune responses induced by P04626 /neu DNA vaccine . DNA vaccine and dendritic cells ( DCs ) -based vaccine have emerged as promising strategies for cancer immunotherapy . P36888 -ligand ( P49771 ) and granulocyte-macrophage-colony-stimulating factor ( GM- P04141 ) have been exploited for the expansion of DC . It was reported previously that combination of plasmid encoding GM- P04141 with P04626 /neu DNA vaccine induced predominantly P01730 (+) T-cell-mediated antitumor immune response . In this study , we investigated the modulation of immune responses by murine P49771 and GM- P04141 , which acted as genetic adjuvants in the forms of bicistronic ( pFLAG ) and monocistronic ( pFL and pGM ) plasmids for P04626 /neu DNA vaccine ( pN-neu ) . Coexpression of P49771 and GM- P04141 significantly enhanced maturation and antigen-presentation abilities of splenic DC . Increased numbers of infiltrating DC at the immunization site , higher interferon-gamma production , and enhanced cytolytic activities by splenocytes were prominent in mice vaccinated with pN-neu in conjunction with pFLAG . Importantly , a potent CD8(+) T-cell-mediated antitumor immunity against bladder tumors naturally overexpressing P04626 /neu was induced in the vaccinated mice . Collectively , our results indicate that murine P49771 and GM- P04141 genes coexpressed by a bicistronic plasmid modulate the class of immune responses and may be superior to those codelivered by two separate monocistronic plasmids as the genetic adjuvants for P04626 /neu DNA vaccine . Inhibition of human brain and RBC acetylcholinesterase ( P22303 ) by heptylphysostigmine ( HPTL ) . Heptylphysostigmine ( HPTL ) , a derivative of the P22303 inhibitor physostigmine ( PHY ) , is under investigation as a therapeutic agent in Alzheimer 's disease . HPTL is active against human RBC P22303 both in vitro and in vivo . Activity of HPTL against human brain has not been documented . We have developed an in vitro assay system using particulate membrane fractions which permits comparison of inhibition and recovery kinetics of human RBC ( primarily globular dimer ) and brain ( primarily globular tetramer ) membrane-bound forms . Under these conditions the HPTLIC50 is similar for the two forms . RBC P22303 inhibition spontaneously reverses in 24 h , as occurs in vivo . In striking contrast , activity of inhibited brain enzyme does not recover on overnight incubation . DDVP-induced inhibition , but not HPTL inhibition , can be reversed by the oxime DB00733 . Some recovery of HPTL inhibition , but not to the level seen with RBC P22303 , occurs on addition of heat-stable fractions from serum or P04141 . Brain enzyme recovers rapidly from PHY in this system . Responses of brain and RBC P22303 to HPTL indicate that these forms are functionally as well as structurally distinct . Since brain inhibition apparently does not spontaneously reverse like RBC inhibition , peripheral measurements of patient responses should be assessed with caution during treatment with HPTL . Src and P61073 are involved in the invasiveness of breast cancer cells with acquired resistance to lapatinib . DB01259 is a dual P00533 and ErbB-2 tyrosine kinase inhibitor that has significantly improved the clinical outcome of ErbB-2-overexpressing breast cancer patients . However , patients inexorably develop mechanisms of resistance that limit the efficacy of the drug . In order to identify potential targets for therapeutic intervention in lapatinib-resistant patients , we isolated , from ErbB-2-overexpressing SK-Br-3 breast cancer cells , the SK-Br-3 Lap-R-resistant subclone , which is able to routinely grow in 1 µM lapatinib . Resistant cells have a more aggressive phenotype compared with parental cells , as they show a higher ability to invade through a matrigel-coated membrane . DB01259 -resistant cells have an increased Src kinase activity and persistent levels of activation of P27361 /2 and AKT compared with parental cells . Treatment with the Src inhibitor saracatinib in combination with lapatinib reduces AKT and P27361 /2 phosphorylation and restores the sensitivity of resistant cells to lapatinib . SK-Br-3 Lap-R cells also show levels of expression of P61073 that are higher compared with parental cells and are not affected by Src inhibition . Treatment with saracatinib or a specific P61073 antibody reduces the invasive ability of SK-Br-3 Lap-R cells , with the two drugs showing cooperative effects . Finally , blockade of Src signaling significantly increases P50591 -induced cell death in SK-Br-3 Lap-R cells . Taken together , our results demonstrate that breast cancer cells with acquired resistance to lapatinib have a more aggressive phenotype compared with their parental counterpart , and that Src signaling and P61073 play an important role in this phenomenon , thus representing potential targets for therapeutic intervention in lapatinib-resistant breast cancer patients . Effects of intense noise exposure on the outer hair cell plasma membrane fluidity . Outer hair cells ( OHCs ) play an important role in cochlear amplification via their length changes ( electromotility ) . A noise-induced cochlear amplification loss leading to a permanent threshold shift ( Q03393 ) was observed without a significant hair cell loss in rats [ Chen , G.D. , Liu , Y. , 2005. Mechanisms of noise-induced hearing loss potentiation by hypoxia. Hear. Res. 200 , 1-9. ] . Since motor proteins are inserted in the OHC lateral membrane , any change in the OHC plasma membrane may result in a loss of OHC electromotility , leading to a loss of cochlear amplification . In this study , the lateral diffusion in the OHC plasma membrane was determined in vitro in guinea pigs by fluorescent recovery after photobleaching ( P42345 ) after an in vivo noise exposure . The lateral diffusion in the OHC plasma membrane demonstrated a length-dependence , which increased as OHC length increased . A reduction in the lateral diffusion was observed in those OHCs with lengths of 50-70 microm after exposure to an 8-kHz octave band noise at 110 dB SPL for 3h . This membrane fluidity change was associated with the selective Q03393 at frequencies around 8 kHz . The reduction of the lateral diffusion in the OHC lateral wall indicated that noise could impair the micromechanics of the OHC lateral wall and might consequently impair OHC electromotility to induce threshold shift . P22888 splicing variants in bovine Leydig cells . The luteinizing hormone receptor ( LHR ) plays a key role in testosterone production through its interaction with the gonadotropins , LH and chorionic gonadotropin . We examined the LHR splicing pattern in bovine Leydig cells ; LH-induced expression of eight cloned splicing variants was detected by real-time PCR . DB00044 applied to cultured Leydig cells resulted in expression of full-length LHR and the A and B isoforms , as well as secretion of testosterone , which first increased , then declined , and then increased further , with increased LH levels . The secretion of testosterone progressively increased with increasing LH , but the expression levels of LHR ( FL , A , and B ) did not increase correspondingly . We conclude that the LHR splicing pattern is complex in bovine Leydig cells , and that expression of full-length LHR and isoforms A and B changes when induced with LH . P23458 activates P40763 activity in non-small-cell lung cancer cells and P05231 neutralizing antibodies can suppress P23458 - P40763 signaling . Members of the signal transducer and activator of transcription ( P35610 ) family of transcription factors are potential targets for the treatment and prevention of cancers including non-small-cell lung cancer . P35610 proteins can be phosphorylated and activated by diverse upstream kinases including cytokine receptors and tyrosine kinases . We examined P35610 protein activation in lung cancer cell lines including those with activating mutations in the P00533 and examined upstream kinases responsible for P40763 phosphorylation and activation using small molecules , antibodies , and RNA interference . We found more pronounced P40763 activation in cells with activating P00533 mutations , yet inhibition of P00533 activity had no effect on P40763 activation . Inhibition of P23458 with small molecules or RNA interference resulted in loss of P40763 tyrosine phosphorylation and inhibition of cell growth . An interleukin-6 neutralizing antibody , siltuximab ( CNTO 328 ) could inhibit P40763 tyrosine phosphorylation in a cell-dependent manner . DB09036 could completely inhibit P40763 tyrosine phosphorylation in H1650 cells , and this resulted in inhibition of lung cancer cell growth in vivo . Combined P00533 inhibition with erlotinib and siltuximab resulted in dual inhibition of both tyrosine and serine P40763 phosphorylation , more pronounced inhibition of P40763 transcriptional activity , and translated into combined effects on lung cancer growth in a mouse model . Our results suggest that P23458 is responsible for P40763 activation in lung cancer cells and that indirect attacks on P23458 - P40763 using an P05231 neutralizing antibody with or without P00533 inhibition can inhibit lung cancer growth in lung cancer subsets . Partial trypsin digestion as an indicator of mis-folding of mutant alanine:glyoxylate aminotransferase and chaperone effects of specific ligands . Study of a spectrum of missense mutants . DB00160 :glyoxylate aminotransferase ( AGT ) is a liver peroxisomal enzyme whose deficiency results in primary hyperoxaluria type 1 ( P78364 ) . More than 75 P78364 mutations are now documented in the AGT gene ( P21549 ) , of which about 50 % are missense . We have previously demonstrated that many such mutants expressed by transcription/translation are subject to generalized degradation by the proteasome and a specific limited trimming by an endogenous DB00171 -independent protease activity . Here , we report the results of partial digestion using trypsin as a mimic for the endogenous non-proteasomal protease and the use of N-terminal protein sequencing to determine the sensitive site . Partial trypsin digestion also provided an indicator of proper folding of the mutant enzyme . For selected mutations the sensitivity to trypsin could be ameliorated by addition of pyridoxal phosphate or aminooxy acetic acid as specific pharmacological chaperones . An acetylcholinesterase inhibitor , eserine , induces long-term depression at P07451 - P00915 synapses in the hippocampus of adult rats . Studies in humans and rodents support a role for muscarinic ACh receptor ( mAChR ) and nicotinic AChR in learning and memory , and both regulate hippocampal synaptic plasticity using complex and often times opposing mechanisms . P22303 ( P22303 ) inhibitors are commonly prescribed to enhance cholinergic signaling in Alzheimer 's disease in hopes of rescuing cognitive function , caused , in part , by degeneration of cholinergic innervation to the hippocampus and cortex . Unfortunately , therapeutic efficacy is moderate and inconsistent , perhaps due to unanticipated mechanisms . M1 mAChRs bidirectionally control synaptic strength at P07451 - P00915 synapses ; weak pharmacological activation using carbachol ( CCh ) facilitates potentiation , whereas strong agonism induces muscarinic long-term depression ( mLTD ) via an P29323 -dependent mechanism . Here , we tested the prediction that accumulation of extracellular ACh via inhibition of P22303 is sufficient to induce LTD at P07451 - P00915 synapses in hippocampal slices from adult rats . Although P22303 inhibition with eserine induces LTD , it unexpectedly does not share properties with mLTD induced by CCh , as reported previously . DB00981 -LTD was prevented by the M3 mAChR-preferring antagonist 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide ( 4-DAMP ) , and pharmacological inhibition of MEK was completely ineffective . Additionally , pharmacological inhibition of p38 MAPK prevents mLTD but has no effect on eserine-LTD . Finally , long-term expression of eserine-LTD is partially dependent on a decrease in presynaptic release probability , likely caused by tonic activation of mAChRs by the sustained increase in extracellular ACh . Thus these findings extend current literature by showing that pharmacological P22303 inhibition causes a prolonged decrease in presynaptic glutamate release at P07451 - P00915 synapses , in addition to inducing a likely postsynaptic form of LTD . P42345 inhibitors and the anti-diabetic biguanide metformin : new insights into the molecular management of breast cancer resistance to the P04626 tyrosine kinase inhibitor lapatinib ( DB01259 ) . The small molecule P04626 tyrosine kinase inhibitor ( TKI ) lapatinib ( DB01259 ) is approved for the therapy of patients with P04626 -positive breast carcinomas who have progressed on trastuzumab ( Herceptin ) . Unfortunately , the efficacy of this P04626 TKI is limited by both primary ( inherent ) and acquired resistance , the latter typically occurring within 12 months of starting therapy . One of the key factors limiting our understanding of the mechanisms involved in lapatinib resistance is the lack of published preclinical models . We herein review lapatinib-refractory models recently developed at the bench and the survival pathways discovered . As hyperactivation of the pharmacologically targetable PI3K/ P42345 /p70S6K1 axis appears to be central to the occurrence of lapatinib resistance , preclinical data showing enhanced antitumour effects when combining lapatinib with P42345 inhibitors ( e.g. , rapamycin analogues and DB00238 -BEZ235 ) highlight the importance of translational work to yield clinically useful regimens capable of delaying or treating lapatinib resistance . The unexpected ability of the anti-type II diabetes drug metformin to inactivate P42345 and decrease p70S6K1 activity further reveals that this biguanide , generally considered non-toxic and remarkably inexpensive , might be considered for new combinatorial lapatinib-based protocols in P04626 -overexpressing breast cancer patients . DB03886 -dependent hyperphenylalaninemia due to deficiency of 6-pyruvoyl tetrahydropterin synthase . We describe the clinical , neurologic , and biochemical findings in 10 patients with 6-pyruvoyl tetrahydropterin synthase ( 6- Q03393 ) deficiency from seven families , all of whom originate from one large tribe in Saudi Arabia . This deficiency presents with severe , early onset of failure to thrive , neurologic deterioration , and morbidity and mortality secondary to repeated episodes of bronchopneumonia or cardiorespiratory abnormalities . The urinary pterin excretion pattern indicates deficient activity of 6- Q03393 , which has been confirmed by direct enzyme assay in red blood cells of three patients . We treated our patients with combined use of tetrahydrobiopterin 20 mg/kg/d , L-dihydroxyphenylalanine 15 mg/kg/d , carbidopa 3.75 mg/kg/d , and L-5-hydroxytryptophan 5 mg/kg/d . Neurologic findings improved significantly in all after 5 to 24 months . Although head circumference and weight returned to the lower limit of normal in four , height normalized only in one of seven patients . Despite an unrestricted diet during combined therapy , blood phenylalanine and urinary excretion of neopterin and biopterin returned to normal . DB06698 ameliorates olanzapine-induced weight gain through modulation of histaminergic , P01303 and AMPK pathways . Olanzapine is widely used to treat schizophrenia and other disorders , but causes adverse obesity and other metabolic side-effects . Both animal and clinical studies have shown that co-treatment with betahistine ( a histaminergic H1 receptor agonist and H3 receptor antagonist ) is effective for ameliorating olanzapine-induced weight gain/obesity . To reveal the mechanisms underlying these effects , this study investigated the effects of co-treatment of olanzapine and betahistine ( O+B ) on expressions of histaminergic H1 receptor ( P35367 ) , AMP-activated protein kinase ( AMPK ) , neuropeptide Y ( P01303 ) , and proopiomelanocortin ( P01189 ) in the hypothalamus associated with reducing olanzapine-induced weight gain . Olanzapine significantly upregulated the mRNA and protein expressions of P35367 , while O+B co-treatment significantly downregulated the P35367 levels , compared to the olanzapine-only treatment group . The P01303 mRNA expression was significantly enhanced by olanzapine , but it was significantly reversed by O+B co-treatment . The hypothalamic P35367 expression was positively correlated with total food intake , and P01303 expression . Olanzapine also increased AMPKα activation measured by the AMPKα phosphorylation ( pAMPKα ) /AMPKα ratio compared with controls , whereas O+B co-treatment decreased the pAMPKα/AMPKα ratio , compared with olanzapine only treatment . The pAMPKα/AMPKα ratio was positively correlated with total food intake and P35367 expression . Although olanzapine administration decreased the P01189 mRNA level , this level was not affected by O+B co-treatment . Therefore , these results suggested that co-treatment with betahistine may reverse olanzapine-induced body weight gain via the P35367 - P01303 and P35367 -pAMPKα pathways . P01133 receptor transactivation and Q96HU1 kinase mediate proteinase-activated receptor-2-induced chloride secretion in intestinal epithelial cells . We examined the stimulus-secretion pathways whereby proteinase-activated receptor 2 ( P55085 ) stimulates Cl(-) secretion in intestinal epithelial cells . SCBN and T84 epithelial monolayers grown on Snapwell supports and mounted in modified Ussing chambers were activated by the P55085 -activating peptides SLIGRL-NH(2) and 2-furoyl-LIGRLO-NH(2) . Short-circuit current ( I(sc) ) was used as a measure of net electrogenic ion transport . Basolateral , but not apical , application of SLIGRL-NH(2) or 2-furoyl-LIGRLO-NH(2) caused a concentration-dependent change in I(sc) that was significantly reduced in Cl(-)-free buffer and by the intracellular Ca(2+) blockers thapsigargin and BAPTA-AM , but not by the Ca(2+) channel blocker verapamil . Inhibitors of PKA ( H-89 ) and P13569 ( glibenclamide ) also significantly reduced P55085 -stimulated Cl(-) transport . P55085 activation was associated with increases in DB02527 and intracellular Ca(2+) . Immunoblot analysis revealed increases in phosphorylation of epidermal growth factor ( P01133 ) receptor ( P00533 ) tyrosine kinase , Src , Pyk2 , P04049 , and P27361 /2 in response to P55085 activation . Pretreatment with inhibitors of cyclooxygenases ( indomethacin ) , tyrosine kinases ( genistein ) , P00533 ( PD-153035 ) , MEK ( PD-98059 or U-0126 ) , and Src ( P50391 ) inhibited SLIGRL-NH(2)-induced increases in I(sc) . Inhibition of Src , but not matrix metalloproteinases , reduced P00533 phosphorylation . Reduced P00533 phosphorylation paralleled the reduction in P55085 -stimulated I(sc) . We conclude that activation of basolateral , but not apical , P55085 induces epithelial Cl(-) secretion via DB02527 - and Ca(2+)-dependent mechanisms . The secretory effect involves P00533 transactivation by Src , leading to subsequent P27361 /2 activation and increased cyclooxygenase activity . Characterisation of the cellular and molecular responses of ovine oocytes and their supporting somatic cells to pre-ovulatory levels of LH and DB00094 during in vitro maturation . The response of Graafian follicles to pre-ovulatory surge levels of DB00094 and LH in vivo triggers the terminal differentiation of granulosa cells and oocyte maturation . In polyovular species , the LH-driven signalling uses the epidermal growth factor ( P01133 ) -like ligands P15514 , O14944 and P35070 to promote oocyte maturation and cumulus expansion . This experimental series used a physiologically relevant ovine in vitro maturation ( IVM ) system to evaluate the impact of exposure to pre-ovulatory levels ( 100 ng/ml ) of LH and DB00094 on ovine cumulus cell expression of P01133 -like ligands in vitro . The serum-free sheep IVM system supported high levels ( 91.4 % ) of gonadotrophin-induced maturation of cumulus-enclosed oocytes and embryo development to the blastocyst stage ( 34.5 % ) . Results were equivalent to a serum-based IVM system ( 85.1 % IVM , 25.8 % blastocyst rate ; P > 0.05 ) but were significantly different ( P < 0.05 ) to serum-free medium without gonadotrophins ( 69.5 % IVM ; 8.0 % blastocyst rate ) . Ovine P35070 was cloned and sequenced . Gonadotrophin-induced P15514 , O14944 , P35070 and P00533 expressions were quantified in cumulus and mural granulosa cells during IVM . A rapid induction of P15514 expression was apparent in both cell types within 30 min of gonadotrophin exposure in vitro . P22888 ( LHR ) was detected in mural cells and P23945 in both cumulus and mural granulosa cells . The data confirm the involvement of P15514 and P00533 during gonadotrophin-induced cumulus expansion , oocyte maturation and the acquisition of developmental competence by sheep oocytes matured in vitro . Targeted treatment of advanced and metastaticbreast cancer with lapatinib . Improved molecular understanding of breast cancer in recent years has led to the discovery of important drug targets such as HER-2 and P00533 . DB01259 is a potent dual inhibitor of HER-2 and P00533 . Preclinical and phase I studies have shown activity with lapatinib in a number of cancers , including breast cancer , and the drug is well tolerated . The main known drug interactions are with paclitaxel and irinotecan . The most significant side-effects of lapatinib are diarrhea and adverse skin events . Rates of cardiotoxicity compare favorably with trastuzumab , a monoclonal antibody against HER-2 . This paper focuses on lapatinib in advanced and metastatic breast cancer , which remains an important therapeutic challenge . Phase II and III studies show activity as monotherapy , and in combination with chemotherapy or hormonal agents . Results from these studies suggest that the main benefit from lapatinib is in the HER-2 positive breast cancer population . Combinations of lapatinib and trastuzumab are also being studied and show encouraging results , particularly in trastuzumab-refractory metastatic breast cancer . DB01259 may have a specific role in treating HER-2 positive CNS metastases . The role of lapatinib as neoadjuvant therapy and in early breast cancer is also being evaluated .

MH_train_1382

MH_train_1382

MH_train_1382

interacts_with DB06212?

multiple_choice

Quantitative analysis of nitric oxide synthase expressed in developing and differentiating rat cerebellum . In order to investigate quantitatively the differentiation and maturation process of granule cells in the postnatal development of rat cerebellum , nitric oxide synthase ( NOS ) activities were determined in the micro-dissected developing cerebellar layers , using our microassay method . NOS activities were increased in the molecular and internal granular layers ( IGLs ) during development and the activity was measurable in the neuroblastic external granular layers ( EGLs ) . Newly devised micro-immunoblot analysis semi-quantitatively showed more amount of endothelial NOS ( P29474 ) and non-negligible amount of neuronal NOS ( P29475 ) in microsamples from EGL , compared with other developing and adult cerebellar layers . P29475 mRNA was also detected using RT-PCR ( reverse transcriptase-polymerase chain reaction ) in the microsamples from this germinal layer . Intraperitoneal administration of DB04223 inhibited NOS activity in vivo and disturbed the layer formation in developing cerebellum . Molecular evolution of the oxytocin-oxytocin receptor system in eutherians . DB00107 ( P01178 ) is a nine-amino-acid peptide hormone that is mainly released at the times of uterine contractions during parturition and milk ejection during lactation , whereas a similar peptide hormone , arginine vasopressin , primarily exerts direct antidiuretic action on the kidney and causes vasoconstriction of the peripheral vessels . The genes coding for these peptides are tandemly located on the same chromosome . A tandem duplication occurring in the common ancestor of jawed vertebrates has been proposed as responsible . In contrast to the two peptide hormones , only one oxytocin receptor ( P30559 ) but three arginine vasopressin receptors ( P37288 , P47901 , and P30518 ) are known ; these receptors probably arose from two rounds of genome duplication in the common ancestor of vertebrates . In this study , we addressed the molecular evolution of the P01178 - P30559 system in eutherians . Our analyses suggest that an amino acid change from isoleucine to lysine on the eighth site ( I8L ) of the peptide , which corresponded to a change from mesotocin to P01178 , had occurred during the common ancestral lineage of eutherians . At around the same time that the emergence of P01178 occurred , functional constraints on the P01178 receptor ( pre- P30559 ) might have relaxed , and a series of nonsynonymous substitutions might have accumulated . Only a few of these nonsynonymous substitutions might have contributed to reestablishing the molecular relationship between the P01178 ligand and its receptor , after which functional constraints on the P30559 were reinstated . Since the P01178 - P30559 system plays an important role in eutherians , the evolution of the P01178 - P30559 system was probably an essential component of the genesis of the eutherian signature . Mechanisms of atrial-selective block of Na⁺ channels by ranolazine : I . Experimental analysis of the use-dependent block . Atrial-selective inhibition of cardiac Na(+) channel current ( I(Na) ) and I(Na)-dependent parameters has been shown to contribute to the safe and effective management of atrial fibrillation . The present study examined the basis for the atrial-selective actions of ranolazine . Whole cell I(Na) was recorded at 15°C in canine atrial and ventricular myocytes and in human embryonic kidney ( P29320 ) -293 cells expressing Q14524 . Tonic block was negligible at holding potentials from -140 to -100 mV , suggesting minimal drug interactions with the closed state . Trains of 40 pulses were elicited over a range of holding potentials to determine use-dependent block . Guarded receptor formalism was used to analyze the development of block during pulse trains . Use-dependent block by ranolazine increased at more depolarized holding potentials , consistent with an interaction of the drug with either preopen or inactivated states , but was unaffected by longer pulse durations between 5 and 200 ms , suggesting a weak interaction with the inactivated state . Block was significantly increased at shorter diastolic intervals between 20 and 200 ms . Responses in atrial and ventricular myocytes and in P29320 -293 cells displayed a similar pattern . DB00243 is an open state blocker that unbinds from closed Na(+) channels unusually fast but is trapped in the inactivated state . Kinetic rates of ranolazine interactions with different states of atrial and ventricular Na(+) channels were similar . Our data suggest that the atrial selectivity of ranolazine is due to a more negative steady-state inactivation curve , less negative resting membrane potential , and shorter diastolic intervals in atrial cells compared with ventricular cells at rapid rates . Excessive fructose intake causes 1,25-(OH)(2)D(3)-dependent inhibition of intestinal and renal calcium transport in growing rats . We recently discovered that chronic high fructose intake by lactating rats prevented adaptive increases in rates of active intestinal Ca(2+) transport and in levels of 1,25-(OH)2D3 , the active form of vitamin D . Since sufficient Ca(2+) absorption is essential for skeletal growth , our discovery may explain findings that excessive consumption of sweeteners compromises bone integrity in children . We tested the hypothesis that 1,25-(OH)2D3 mediates the inhibitory effect of excessive fructose intake on active Ca(2+) transport . First , compared with those fed glucose or starch , growing rats fed fructose for 4 wk had a marked reduction in intestinal Ca(2+) transport rate as well as in expression of intestinal and renal Ca(2+) transporters that was tightly associated with decreases in circulating levels of 1,25-(OH)2D3 , bone length , and total bone ash weight but not with serum parathyroid hormone ( PTH ) . Dietary fructose increased the expression of 24-hydroxylase ( Q07973 ) and decreased that of 1α-hydroxylase ( O15528 ) , suggesting that fructose might enhance the renal catabolism and impair the synthesis , respectively , of 1,25-(OH)2D3 . Serum Q9GZV9 , which is secreted by osteocytes and inhibits O15528 expression , was upregulated , suggesting a potential role of bone in mediating the fructose effects on 1,25-(OH)2D3 synthesis . Second , 1,25-(OH)2D3 treatment rescued the fructose effect and normalized intestinal and renal Ca(2+) transporter expression . The mechanism underlying the deleterious effect of excessive fructose intake on intestinal and renal Ca(2+) transporters is a reduction in serum levels of 1,25-(OH)2D3 . This finding is significant because of the large amounts of fructose now consumed by Americans increasingly vulnerable to Ca(2+) and vitamin D deficiency . Autosomal-dominant hypophosphatemic rickets ( P30518 ) mutations stabilize Q9GZV9 . BACKGROUND : The gene for the renal phosphate wasting disorder autosomal-dominant hypophosphatemic rickets ( P30518 ) is Q9GZV9 , which encodes a secreted protein related to the fibroblast growth factors ( FGFs ) . We previously detected missense mutations R176Q , R179W , and R179Q in Q9GZV9 from P30518 kindreds . The mutations replace R residues within a subtilisin-like proprotein convertase ( Q969E3 ) cleavage site 176RHTR-179 ( RXXR motif ) . The goal of these studies was to determine if the P30518 mutations lead to protease resistance of Q9GZV9 . METHODS : The P30518 mutations were introduced into human Q9GZV9 cDNA clones with or without an N-terminal FLAG tag by site-directed mutagenesis and were transiently transfected into HEK293 cells . Protein expression was determined by Western analyses . RESULTS : Antibodies directed toward the C-terminal portion of Q9GZV9 revealed that the native Q9GZV9 protein resolved as 32 kD and 12 kD species in HEK293 conditioned media ; however , the three mutated proteins were detected only as the 32 kD band . An N-terminal FLAG-tagged native Q9GZV9 resolved as two bands of 36 kD and 26 kD when detected with a FLAG antibody , whereas the R176Q mutant resolved primarily as the 36 kD protein species . Cleavage of Q9GZV9 was not enhanced by extracellular incubation of Q9GZV9 with HEK293 cells . Native and mutant FGF-23s bound heparin . CONCLUSIONS : Q9GZV9 proteins containing the P30518 mutations are secreted , and produce polypeptides less sensitive to protease cleavage than wild-type Q9GZV9 . Therefore , the P30518 mutations may protect Q9GZV9 from proteolysis , thereby potentially elevating circulating concentrations of Q9GZV9 and leading to phosphate wasting in P30518 patients . P06401 modulator DB05366 down-regulates vascular endothelial growth factor , adrenomedullin and their receptors and modulates progesterone receptor content in cultured human uterine leiomyoma cells . BACKGROUND : This study was conducted to evaluate the effects of graded concentrations ( 10(-8) , 10(-7) and 10(-6) M ) of progesterone receptor ( PR ) modulator DB05366 on the protein contents of PR , of vascular endothelial growth factor ( P15692 ) , adrenomedullin ( P35318 ) and their receptors in cultured human uterine leiomyoma and matching myometrial cells . METHODS : PR-A , PR-B , P15692 , P49765 , P15692 receptor ( VEGFR ) -1 , P35968 , P35318 and P35318 receptor ( O15218 ) contents were assessed by Western blot analysis . RESULTS : Treatment with 100 ng/ml progesterone increased P15692 , P49765 and P35318 contents in cultured leiomyoma cells and normal myometrial cells . The concomitant treatment with 10(-6) M DB05366 significantly decreased the progesterone-induced P15692 , P49765 and P35318 contents in cultured leiomyoma cells but not in normal myometrial cells . DB05366 treatment alone decreased P17948 , P35968 and O15218 contents in cultured leiomyoma cells but not in normal myometrial cells . DB05366 treatment increased PR-A and decreased PR-B contents in cultured leiomyoma cells in a dose-dependent manner compared with untreated cultures , whereas no significant changes in PR isoform contents were observed in normal myometrial cells . CONCLUSIONS : These results suggest that DB05366 down-regulates P15692 , P35318 and their receptor contents and modulates PR isoform contents in cultured leiomyoma cells in a cell-type-specific manner . 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 . Functional autoradiography of neuropeptide Y Q03519 and P49146 subtypes in rat brain using agonist stimulated [35S]GTPgammaS binding . Neuropeptide Y , one of the most abundant brain peptides , has been found to modulate several important biological functions via a family of G-protein coupled receptors . To investigate the localization of functional P01303 receptor subtypes in the rat brain , we performed agonist-induced [35S]GTPgammaS autoradiography . The Q03519 /Y4/Y5 agonist DB00149 (31) , Pro(34)- P01303 increased [35S]GTPgammaS binding in several brain areas with a regional distribution consistent with that produced when labeling adjacent sections with [ 125I ] - DB00149 (31) , Pro(34)- P10082 . The Q03519 selective antagonist BIBP3226 antagonized the DB00149 (31) , Pro(34)- P01303 stimulated increase in [35S]GTPgammaS binding in all areas examined . The P28062 agonist P06681 - P01303 stimulated [35S]GTPgamma binding in numerous brain areas with a regional distribution similar to the binding observed with [ 125I ] - DB05004 . No increase in [35S]GTPgammaS binding above basal was observed in any brain area evaluated using Y4 and Y5 selective agonists . This study demonstrates abundant Q03519 and P49146 activation in the rat brain , while evidence for functional Y4 and Y5 receptors was not observed . Evolution of spatially coexpressed families of type-2 vomeronasal receptors in rodents . The vomeronasal organ ( VNO ) is an olfactory structure for the detection of pheromones . VNO neurons express three groups of unrelated G-protein-coupled receptors . Type-2 vomeronasal receptors ( V2Rs ) are specifically localized in the basal neurons of the VNO and are believed to sense protein pheromones eliciting specific reproductive behaviors . In murine species , V2Rs are organized into four families . Family-ABD V2Rs are expressed monogenically and coexpress with family-C V2Rs of either subfamily C1 ( V2RC1 ) or subfamily P06681 ( V2RC2 ) , according to a coordinate temporal diagram . Neurons expressing the phylogenetically ancient V2RC1 coexpress family-BD V2Rs or a specific group of subfamily-A V2Rs ( V2RA8-10 ) , whereas a second neuronal subset ( V2RC2-positive ) coexpresses a recently expanded group of five subfamily-A V2Rs ( V2RA1-5 ) along with vomeronasal-specific Major Histocompatibility Complex molecules ( H2-Mv ) . Through database mining and Sanger sequencing , we have analyzed the onset , diversification , and expansion of the P30518 -families throughout the phylogeny of Rodentia . Our results suggest that the separation of V2RC1 and V2RC2 occurred in a Cricetidae ancestor in coincidence with the evolution of the H2-Mv genes ; this phylogenetic event did not correspond with the origin of the coexpressing V2RA1-5 genes , which dates back to an ancestral myomorphan lineage . Interestingly , the evolution of receptors within the V2RA1-5 group may be implicated in the origin and diversification of some of the P30518 putative cognate ligands , the exocrine secreting peptides . The establishment of V2RC2 , which probably reflects the complex expansion and diversification of family-A V2Rs , generated receptors that have probably acquired a more subtle functional specificity . Neuronal and glial response in the rat hypothalamus-neurohypophysis complex with streptozotocin-induced diabetes . This study was aimed to examine the neuronal and glial response in the hypothalamus and neurohypophysis of rats with streptozotocin-induced diabetes . At various time intervals after induction of diabetes the neurons in the paraventricular- ( PVN ) and supraoptic- ( P18583 ) nucleus showed upregulated arginine vasopressin ( AVP ) and oxytocin ( P01178 ) immunoexpression , being most pronounced at 2 weeks . Concomitant to this was the hypertrophy of PVN and P18583 neurons . Q05586 , which was constitutively and moderately expressed in normal rats , was markedly augmented , being most intense at 4 months . This coincided with the expression of neuronal nitric oxide synthase ( P29475 ) . Contrary to this , the expression of GluR2/3 was progressively downregulated , so that it was hardly detected at 4 months . Both astrocytes and microglia marked by anti- P14136 and OX-42 , respectively , appeared activated . In pars nervosa , the projection target of the axon terminals of PVN and P18583 neurons , massive axons and terminals ( Herring bodies ) laden with neurosecretions were observed in diabetic rats . Colocalization study showed that the neurosecretions were internalized by activated pituicytes and microglia associated with the axons . The present results suggest that the neurosecretion of PVN and P18583 neurons is enhanced in diabetes . This is coupled by upregulation of Q05586 and P29475 but downregulation of GluR2/3 . It is speculated that the glutamate receptors and NO are linked to overactivation of PVN and P18583 neurons leading ultimately to cell death of some of them . The pituicytes and microglia in pars nervosa would help to modulate the release of neurosecretion . Coordinated regulation of insulin signaling by the protein tyrosine phosphatases P18031 and P17706 . The protein tyrosine phosphatase P18031 is a negative regulator of insulin signaling and a therapeutic target for type 2 diabetes . Our previous studies have shown that the closely related tyrosine phosphatase P17706 might also contribute to the regulation of insulin receptor ( IR ) signaling in vivo ( S. Galic , M. Klingler-Hoffmann , M. T. Fodero-Tavoletti , M. A. Puryer , T. C. Meng , N. K. Tonks , and T. Tiganis , Mol. Cell. Biol. 23:2096-2108 , 2003 ) . Here we show that P18031 and P17706 function in a coordinated and temporally distinct manner to achieve an overall regulation of IR phosphorylation and signaling . Whereas insulin-induced phosphatidylinositol 3-kinase/Akt signaling was prolonged in both P17706 -/- and P18031 -/- immortalized mouse embryo fibroblasts ( MEFs ) , mitogen-activated protein kinase P27361 /2 signaling was elevated only in P18031 -null MEFs . By using phosphorylation-specific antibodies , we demonstrate that both IR beta-subunit Y1162/Y1163 and Y972 phosphorylation are elevated in P18031 -/- MEFs , whereas Y972 phosphorylation was elevated and Y1162/Y1163 phosphorylation was sustained in P17706 -/- MEFs , indicating that P18031 and P17706 differentially contribute to the regulation of IR phosphorylation and signaling . Consistent with this , suppression of P17706 protein levels by RNA interference in P18031 -/- MEFs resulted in no change in P27361 /2 signaling but caused prolonged Akt activation and Y1162/Y1163 phosphorylation . These results demonstrate that P18031 and P17706 are not redundant in insulin signaling and that they act to control both common as well as distinct insulin signaling pathways in the same cell . Phosphorylation and recycling kinetics of G protein-coupled receptors . The rate of ligand-induced phosphorylation of the V2 and V1a vasopressin receptors was characterized in P29320 293 cells . Both receptors were phosphorylated predominantly by GRKs , and the V1a receptor was also phosphorylated by protein kinase C regardless of the presence or absence of ligand . Phosphorylation of the P37288 catalyzed by GRKs reached maximal values at the shortest measured time : 15 seconds , and decayed rapidly with a t1/2 of 6 min in the continuous presence of AVP . In agreement with the hypothesis that dephosphorylation must precede receptor recycling to the cell surface , the P37288 returned rapidly to the cell surface after removal of the hormone from the medium . Phosphate incorporation into the P30518 proceeded at a slower pace , and the internalized phosphorylated receptor failed to recycle to the cell surface and retained its phosphate for a long time in the presence or absence of ligand . A single mutation in the carboxy terminus of the P30518 accelerated de-phosphorylation of the protein and conferred recycling properties to the P30518 . These experiments provided molecular evidence for the hypothesis that internalization is required for de-phosphorylation and recycling of reactivated G protein coupled receptors to the cell surface . P11926 activity in human endometrium and endometrial cancer cells . P11926 ( ODC ) activities were significantly higher in proliferative endometrium during the estrogen-dominated follicular phase of the menstrual cycle than in secretory endometrium after the formation of the progesterone-secreting corpus luteum . The enzymatic activity was increased about fivefold by renewal of the medium during incubations of endometrial fragments or isolated endometrial glands . Endometrial adenocarcinoma cells ( O14777 -1 , O14777 -50 ) , both in monolayers and suspension , also responded to medium renewal by increasing ODC activity about 10-fold after 4 h , with subsequent reduction to control levels after 7 h . These effects were blocked by actinomycin D and cycloheximide . Endometrial stromal cells exhibited highly variable ODC activities at different passages . Difluoromethylornithine ( DB06243 ) and sodium molybdate had marked antiproliferative effects in O14777 -50 cultures , reducing cell numbers to 10 to 20 % of control values 11 d after plating and inhibiting ODC activity by approximately 80 % on Day 7 . The antiproliferative effect of DB06243 , but not that of molybdate , was reversed by 10 microM putrescine , the product of ODC activity . In contrast to DB06243 , molybdate had no effect on ODC activity of cell homogenates . Molybdate did not elicit antizyme formation in O14777 -50 cells under conditions in which putrescine did . These results indicate that ODC activity , present in both epithelial and stromal cells , as shown analytically and also by autoradiography after labeling with [3H] DB06243 , may be related to cell proliferation in vivo and that proliferation of human endometrial cancer cells in culture can be arrested by DB06243 and by molybdate . P03952 promotes epidermal growth factor receptor transactivation and signaling in vascular smooth muscle through direct activation of protease-activated receptors . The kallikrein-kinin system , along with the interlocking renin-angiotensin system , is a key regulator of vascular contractility and injury response . The principal effectors of the kallikrein-kinin system are plasma and tissue kallikreins , proteases that cleave high molecular weight kininogen to produce bradykinin . Most of the cellular actions of kallikrein ( KK ) are thought to be mediated by bradykinin , which acts via G protein-coupled B1 and B2 bradykinin receptors on VSMCs and endothelial cells . Here , we find that primary aortic vascular smooth muscle but not endothelial cells possess the ability to activate plasma prekallikrein . Surprisingly , exposing VSMCs to prekallikrein leads to activation of the P27361 /2 mitogen-activated protein kinase cascade via a mechanism that requires kallikrein activity but does not involve bradykinin receptors . In transfected HEK293 cells , we find that plasma kallikrein directly activates G protein-coupled protease-activated receptors ( PARs ) 1 and 2 , which possess consensus kallikrein cleavage sites , but not PAR4 . In vascular smooth muscles , KK stimulates ADAM ( a disintegrin and metalloprotease ) 17 activity via a PAR1/2 receptor-dependent mechanism , leading sequentially to release of the endogenous P78536 substrates , amphiregulin and tumor necrosis factor-α , metalloprotease-dependent transactivation of epidermal growth factor receptors , and metalloprotease and epidermal growth factor receptor-dependent P27361 /2 activation . These results suggest a novel mechanism of bradykinin-independent kallikrein action that may contribute to the regulation of vascular responses in pathophysiologic states , such as diabetes mellitus . Desmopressin ( DB00035 ) induces NO production in human endothelial cells via V2 receptor- and DB02527 -mediated signaling . The hemostatic agent desmopressin ( DB00035 ) also has strong vasodilatory effects . DB00035 is a selective agonist for the vasopressin V2 receptor ( P30518 ) , which is coupled to DB02527 -dependent signaling . DB00035 -induced vasodilation may be due to endothelial NO synthase ( P29474 ) activation . This hypothesis implies DB02527 -mediated P29474 activation . It also implies wide extrarenal , endothelial P30518 expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 -raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 enzymatic activity , in a partly calcium-independent manner . DB02527 -mediated P29474 activation is associated with phosphorylation of residue Ser1177 , in a phosphatidyl inositol 3-kinase ( PI3K ) -independent manner . HUVECs do not express P30518 . However , after heterologous P30518 expression , DB00035 induces DB02527 -dependent P29474 activation via Ser1177 phosphorylation . We have previously found P30518 expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 and other DB02527 -raising agents can activate P29474 via PI3K-independent Ser1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 -induced vasodilation . Biomimetic growth of bone-like apatite via simulated body fluid on hydroxyethyl cellulose/polyvinyl alcohol electrospun nanofibers . In this study , randomly oriented hydroxyethyl cellulose/polyvinyl alcohol ( O14777 / P32926 ) nanofibers were fabricated by electrospinning . The blend solutions of O14777 / P32926 with different weight ratio of O14777 to P32926 were prepared using water as solvent to fabricate nanofibers . These nanofibrous scaffolds were coated with bone-like apatite by immersing into 10x simulated body fluid ( P52747 ) for different time periods . The morphology and structure of the nanofibers were characterized by SEM , FTIR and DSC . FESEM-EDS and FTIR analysis were used to confirm the deposition of apatite on the surface of nanofibers . The results of this study suggest that this apatite coated nanofibrous scaffolds could be a suitable biomaterial for bone tissue engineering . [ Synthesis of ( 2'-bromo-4 ' , 5'-dimethoxy-phenyl ) -(2,3-dibromo-4,5-dimethoxy-phenyl)-methane as P18031 inhibitor ] . OBJECTIVE : To synthesize (2'-bromo-4',5'-dimethoxy-phenyl)- ( 2,3- dibromo-4,5-dimethoxy-phenyl ) -methane ( 6 ) as protein tyrosine phosphatase 1B ( P18031 ) inhibitor . METHOD : DB04088 was synthesized by Friedel-Crafts reaction , bromination and decarbonylation and screened inhibitory activity against P18031 by the colorimetric assay . The structure of synthetic intermediates and target product were identified on the basis of spectral analysis . RESULT : DB04088 was synthesized successfully in four steps and evaluated for its P18031 inhibitory activity , the screening result shown that compound 6 displayed good inhibitory activity against P18031 . CONCLUSION : The target compound 6 was synthesized with the overall yield of 20 % , which was a new compound and shown good inhibitory activity against P18031 ( inhibition 40.16 % at 5 mg x L(-1) ) . P06850 family of peptides regulates intestinal angiogenesis . BACKGROUND & AIMS : The corticotrophin-releasing hormone ( P06850 ) family of peptides modulates intestinal inflammation and the P06850 receptor 2 ( Q13324 ) suppresses postnatal angiogenesis in mice . We investigated the functions of P34998 and Q13324 signaling during intestinal inflammation and angiogenesis . METHODS : The activities of P34998 and Q13324 were disrupted by genetic deletion in mice or with selective antagonists . A combination of in vivo , ex vivo , and in vitro measures of angiogenesis were used to determine their activity . P34998 (-/-) mice and Q13324 (-/-) mice with dextran sodium sulfate-induced colitis were analyzed in comparison with wild-type littermates ( controls ) . RESULTS : Colitis was significantly reduced in mice in which P34998 activity was disrupted by genetic deletion or with an antagonist , determined by analyses of survival rate , weight loss , histological scores , and cytokine production . Inflammation was exacerbated in mice in which Q13324 activity was inhibited by genetic deletion or with an antagonist , compared with controls . The inflamed intestines of P34998 (-/-) mice had reduced microvascular density and expression of vascular endothelial growth factor ( P15692 ) -A , whereas the intestines of Q13324 (-/-) mice had increased angiogenesis and P15692 levels . An antagonist of P35968 activity alleviated colitis in Q13324 (-/-) mice . Ex vivo aortic vessel outgrowth was reduced when P34998 was deficient but increased when Q13324 was deficient . The P34998 preferred agonist P06850 stimulated tube formation , proliferation , and migration of cultured intestinal microvascular endothelial cells by phosphorylating Akt , whereas the specific Q13324 agonist Q969E3 had opposite effects . CONCLUSION : P34998 promotes intestinal inflammation , as well as endogenous and inflammatory angiogenesis whereas Q13324 inhibits these activities . Receptor to glutamate DB01221 -type : the functional diversity of the nr1 isoforms and pharmacological properties . DB00142 ( DB00142 ) is the major excitatory neurotransmitter in the central nervous system , and interacts with two classes of receptor : metabotropic and ionotropic receptors . Ionotropic receptors are divided according to the affinity of their specific agonists : Nmethyl- D-aspartate ( DB01221 ) , amino acid-3-hydroxy-5-methyl-4-isoxazole acid ( AMPA ) and kainic acid ( KA ) . DB01221 receptors ( DB01221 -R ) are macromolecular structures that are formed by different combinations of subunits : Q05586 ( Q9UHB4 ) , NMDAR2 ( NR2 ) and NMDAR3 ( Q13224 ) . The study of this receptor has aroused great interest , partly due to its role in synaptic plasticity but mainly because of its permeability to the Ca(2+) ion . This review examines the molecular composition of DB01221 -R and the variants of Q9UHB4 subunit editing in association with NR2 subunit dimers , which form the main components of this receptor . Their composition , structure , function and distinct temporal and spatial expression patterns demonstrate the versatility and diversity of functionally different isoforms of Q9UHB4 subunits and the various pharmacological properties of the NR2 subunit . Finally , the involvement of DB01221 -R in the excitotoxicity phenomenon , as well as , its expression changes under these conditions as neuronal response are also discussed . Selective measurement of white matter and gray matter diffusion trace values in normal human brain . The trace of the diffusion tensor ( or simply the trace ) is diagnostically valuable for detecting acute ischemic lesions . A number of studies indicate that the trace of human gray matter ( GM ) and white matter ( WM ) are quite similar . This is somewhat surprising considering the different cellular environments of GM and WM . It is possible that partial volume averaging ( P32926 ) effects between GM and WM , inherent in many of the ultrafast imaging sequences used for diffusion measurements , are responsible for this observation . In order to minimize P32926 effects , the trace values of GM and WM have been selectively measured by implementing double inversion recovery ( P30518 ) echo planar imaging ( P10646 ) pulse sequences . Results on six normal volunteers indicate that the trace values of WM and GM are not statistically different . The V2 vasopressin receptor stimulates P27361 /2 activity independently of heterotrimeric G protein signalling . The V2 vasopressin receptor ( P30518 ) activates the mitogen activated protein kinases ( MAPK ) P27361 /2 through a mechanism involving the scaffolding protein beta arrestin . Here we report that this activating pathway is independent of G alpha s , G alpha i , G alpha q or G betagamma and that the P30518 -mediated activation of G alpha s inhibits P27361 /2 activity in a DB02527 /PKA-dependent manner . In the HEK293 cells studied , the beta arrestin-promoted activation was found to dominate over the PKA-mediated inhibition of the pathway , leading to a strong vasopressin-stimulated P27361 /2 activation . Despite the strong MAPK activation and in contrast with other GPCR , P30518 did not induce any significant increase in DNA synthesis , consistent with the notion that the stable interaction between P30518 and beta arrestin prevents signal propagation to the nucleus . Beta arrestin was found to be essential for the P27361 /2 activation , indicating that the recruitment of the scaffolding protein is necessary and sufficient to initiate the signal in the absence of any other stimulatory cues . Based on the use of selective pharmacological inhibitors , dominant negative mutants and siRNA , we conclude that the beta arrestin-dependent activation of P27361 /2 by the P30518 involves c-Src and a metalloproteinase-dependent trans-activation event . These findings demonstrate that beta arrestin is a genuine signalling initiator that can , on its own , engage a MAPK activation machinery upon stimulation of a GPCR by its natural ligand . DB05311 ( DX-88 ) , a plasma kallikrein inhibitor for the treatment of hereditary angioedema and the prevention of blood loss in on-pump cardiothoracic surgery . BACKGROUND : P03952 plays a major role in the contact ( kallikrein-kinin ) cascade producing bradykinin . Bradykinin is a vasodilator , which increases vascular permeability , activates inflammation and produces pain . P03952 is also crosslinked to the coagulation system and the complement cascade . OBJECTIVE : DB05311 ( DX-88 ) is a potent and specific inhibitor of plasma kallikrein . DB05311 is a recombinantly produced and engineered small protein based on the first Kunitz domain of human tissue factor pathway inhibitor . It was identified through phage display technology . METHODS : The search terms ' ecallantide ' , ' DX-88 ' and ' hereditary angioedema ' were entered into Pubmed/Medline , ClinicalTrials and Google . RESULTS/CONCLUSION : At present , the drug is being studied for two major indications . First , the results for the treatment of hereditary angioedema are promising . Second , a prospective randomised multi-centre trial for the reduction of blood loss during on-pump cardiothoracic surgery will be terminated in October 2008 . Vitamin D metabolism , mechanism of action , and clinical applications . DB00169 is made in the skin from 7-dehydrocholesterol under the influence of UV light . DB00153 ( ergocalciferol ) is derived from the plant sterol ergosterol . Vitamin D is metabolized first to 25 hydroxyvitamin D ( 25OHD ) , then to the hormonal form 1,25-dihydroxyvitamin D ( 1,25(OH)2D ) . Q6VVX0 is the most important 25-hydroxylase ; O15528 is the key 1-hydroxylase . Both 25OHD and 1,25(OH)2D are catabolized by Q07973 . 1,25(OH)2D is the ligand for the vitamin D receptor ( P11473 ) , a transcription factor , binding to sites in the DNA called vitamin D response elements ( VDREs ) . There are thousands of these binding sites regulating hundreds of genes in a cell-specific fashion . P11473 -regulated transcription is dependent on comodulators , the profile of which is also cell specific . Analogs of 1,25(OH)2D are being developed to target specific diseases with minimal side effects . This review will examine these different aspects of vitamin D metabolism , mechanism of action , and clinical application .

MH_train_1383

MH_train_1383

MH_train_1383

interacts_with DB01171?

multiple_choice

The receptor for urokinase regulates O60603 mediated inflammatory responses in neutrophils . The urokinase-type plasminogen activator receptor ( Q03405 ) , a glycosylphosphatidylinositol ( P06744 ) anchored membrane protein , regulates urokinase ( uPA ) protease activity , chemotaxis , cell-cell interactions , and phagocytosis of apoptotic cells . Q03405 expression is increased in cytokine or bacteria activated cell populations , including macrophages and monocytes . However , it is unclear if Q03405 has direct involvement in the response of inflammatory cells , such as neutrophils and macrophages , to Toll like receptor ( TLR ) stimulation . In this study , we found that Q03405 is required for optimal neutrophil activation after O60603 , but not O00206 stimulation . We found that the expression of P01375 -α and P05231 induced by O60603 engagement in Q03405 -/- neutrophils was less than that in Q03405 +/+ ( WT ) neutrophils . Pretreatment of neutrophils with PI- P98160 , which cleaves P06744 moieties , significantly decreased O60603 induced expression of P01375 -α in WT neutrophils , but demonstrated only marginal effects on P01375 -α expression in PAM treated Q03405 -/- neutrophils . IκB-α degradation and NF-κB activation were not different in Q03405 -/- or WT neutrophils after O60603 stimulation . However , Q03405 is required for optimal p38 MAPK activation after O60603 engagement . Consistent with the in vitro findings that Q03405 modulates O60603 engagement induced neutrophil activation , we found that pulmonary and systemic inflammation induced by O60603 , but not O00206 stimulation is reduced in Q03405 -/- mice compared to WT counterparts . Therefore , our data suggest that neutrophil associated Q03405 could be a potential target for treating acute inflammation , sepsis , and organ injury related to severe bacterial and other microbial infections in which O60603 engagement plays a major role . Inhibition of the invasion capacity of carcinoma cells by DB05476 , 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 , a novel 3-amidinophenylalanine-based inhibitor of the uPA system , at inhibiting the invasive capacity of carcinoma cells . First , Q03405 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 expression levels correlated positively with invasion capacity , which could be significantly inhibited by DB05476 . 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 . Thus , our results demonstrate the potential of DB05476 in vitro as a promising adjuvant antimetastatic therapy of carcinomas . Multiple cholinergic nicotinic receptor genes affect nicotine dependence risk in African and European Americans . Several independent studies show that the chromosome 15q25.1 region , which contains the P30532 - P32297 - P30926 gene cluster , harbors variants strongly associated with nicotine dependence , other smoking behaviors , lung cancer and chronic obstructive pulmonary disease . We investigated whether variants in other cholinergic nicotinic receptor subunit ( CHRN ) genes affect the risk of nicotine dependence in a new sample of African Americans ( AAs ) ( N = 710 ) . We also analyzed this AA sample together with a European American ( EA ) sample ( N = 2062 , 1608 of which have been previously studied ) , allowing for differing effects in the two populations . Cases are current nicotine-dependent smokers and controls are non-dependent smokers . Variants in or near Q07001 - P07510 , P36544 and Q9GZZ6 show modest association with nicotine dependence risk in the AA sample . In addition , P43681 , Q05901 - Q15825 and P11230 show association in at least one population . P07510 and P43681 harbor single nucleotide polymorphisms ( SNPs ) that have opposite directions of effect in the two populations . In each of the population samples , these loci substantially increase the trait variation explained , although no loci meet Bonferroni-corrected significance in the AA sample alone . The trait variation explained by three key associated SNPs in P30532 - P32297 - P30926 is 1.9 % in EAs and also 1.9 % in AAs ; this increases to 4.5 % in EAs and 7.3 % in AAs when we add six variants representing associations at other CHRN genes . Multiple nicotinic receptor subunit genes outside chromosome 15q25 are likely to be important in the biological processes and development of nicotine dependence , and some of these risks may be shared across diverse populations . Discovery of DB05130 , a Potent , Selective , and Orally Bioavailable hCCR2 Antagonist . We report the identification of 13 ( DB05130 ) as a potent human P41597 ( hCCR2 ) antagonist . DB05130 exhibited an IC50 of 3.7 nM in antagonism of monocyte chemoattractant protein-1 binding to hCCR2 , 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 exhibited a pharmacokinetic profile suitable for once-a-day dosing ( T 1/2 = 15 h ) . Nrf2 is a critical modulator of the innate immune response in a model of uveitis . Uveitis is an inflammatory condition that can lead to blindness . It is therefore important to understand the pathophysiology against which to develop targeted therapy . Herein , we tested whether the oxidant-responsive transcription factor Nrf2 is involved in regulating the innate immune response and oxidative damage in the LPS uveitis model . As shown by dihydroethidium staining , intraperitoneally injected LPS increased reactive oxygen species in the retina and iris-ciliary body of Nrf2+/+ and Nrf2-/- mice . After LPS injection , P05362 , P05231 , P01375 , P35354 , P35228 , and P13500 mRNAs were increased more in the retina and iris-ciliary body of Nrf2-/- than in those of Nrf2+/+ mice . NQO-1 and P48507 , two Nrf2-responsive antioxidant enzymes , had reduced expression in Nrf2+/+ retinas after LPS injection , but no change in expression in Nrf2-/- mice . The number of FITC-Con A-labeled leukocytes adherent to the retinal vascular endothelium increased after LPS treatment in both Nrf2+/+ and Nrf2-/- mice compared to control injections , with more adherent leukocytes in Nrf2-/- than in Nrf2+/+ mice . Pretreatment with the Nrf2 activator 1-(2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oyl)imidazole increased antioxidant gene expression in the retina , reduced inflammatory mediator expression , and reduced leukocyte adherence to retinal vasculature after LPS treatment in Nrf2+/+ mice , but had no effect on Nrf2-/- mice . Treatment targeting the Nrf2 pathway may be a new therapy for uveitis . Hemizygosity of transsulfuration genes confers increased vulnerability against acetaminophen-induced hepatotoxicity in mice . The key mechanism for acetaminophen hepatotoxicity is cytochrome P450 ( CYP ) -dependent formation of N-acetyl-p-benzoquinone imine , a potent electrophile that forms protein adducts . Previous studies revealed the fundamental role of glutathione , which binds to and detoxifies N-acetyl-p-benzoquinone imine . Glutathione is synthesized from cysteine in the liver , and DB06151 is used as a sole antidote for acetaminophen poisoning . Here , we evaluated the potential roles of transsulfuration enzymes essential for cysteine biosynthesis , cystathionine β-synthase ( P35520 ) and cystathionine γ-lyase ( CTH ) , in acetaminophen hepatotoxicity using hemizygous ( Cbs(+/-) or Cth(+/-) ) and homozygous ( Cth(-/-) ) knockout mice . At 4 h after intraperitoneal acetaminophen injection , serum alanine aminotransferase levels were highly elevated in Cth(-/-) mice at 150 mg/kg dose , and also in Cbs(+/-) or Cth(+/-) mice at 250 mg/kg dose , which was associated with characteristic centrilobular hepatocyte oncosis . Hepatic glutathione was depleted while serum malondialdehyde accumulated in acetaminophen-injected Cth(-/-) mice but not wild-type mice , although glutamate-cysteine ligase ( composed of catalytic [ P48506 ] and modifier [ P48507 ] subunits ) became more activated in the livers of Cth(-/-) mice with lower Km values for DB00151 and DB00142 . Proteome analysis using fluorescent two-dimensional difference gel electrophoresis revealed 47 differentially expressed proteins after injection of 150 mg acetaminophen/kg into Cth(-/-) mice ; the profiles were similar to 1000 mg acetaminophen/kg-treated wild-type mice . The prevalence of Cbs or Cth hemizygosity is estimated to be 1:200-300 population ; therefore , the deletion or polymorphism of either transsulfuration gene may underlie idiosyncratic acetaminophen vulnerability along with the differences in Cyp , Gclc , and Gclm gene activities . 5-hydroxytryptamine receptors in the human cardiovascular system . The human cardiovascular system is exposed to plasma 5-hydroxytryptamine ( 5-HT , serotonin ) , usually released from platelets . 5-HT can produce harmful acute and chronic effects . The acute cardiac effects of 5-HT consist of tachycardia ( preceded on occasion by a brief reflex bradycardia ) , increased atrial contractility and production of atrial arrhythmias . Acute inotropic , lusitropic and arrhythmic effects of 5-HT on human ventricle become conspicuous after inhibition of phosphodiesterase ( PDE ) activity . Human cardiostimulation is mediated through Q13639 receptors . Atrial and ventricular PDE3 activity exerts a protective role against potentially harmful cardiostimulation . Chronic exposure to high levels of 5-HT ( from metastatic carcinoid tumours ) , the anorectic drug fenfluramine and its metabolites , as well as the ecstasy drug 3,4-methylenedioxymethamphetamine ( DB01454 ) and its metabolite 3,4-methylenedioxyamphetamine ( MDA ) are associated with proliferative disease and thickening of cardiac valves , mediated through P41595 receptors . P41595 receptors have an obligatory physiological role in murine cardiac embryology but whether this happens in humans requires research . Congenital heart block ( CHB ) is , on occasion , associated with autoantibodies against Q13639 receptors . Acute vascular constriction by 5-HT is usually shared by P28222 and 5- Q13049 receptors , except in intracranial arteries which constrict only through P28222 receptors . Both P28222 and 5- Q13049 receptors can mediate coronary artery spasm but only P28222 receptors appear involved in coronary spasm of patients treated with triptans or with Prinzmetal angina . 5- Q13049 receptors constrict the portal venous system including oesophageal collaterals in cirrhosis . Chronic exposure to 5-HT can contribute to pulmonary hypertension through activation of constrictor P28222 receptors and proliferative P41595 receptors , and possibly through direct intracellular effects . Modulation of cytokine production and enhancement of cell viability by Q9NYK1 and Q9NR96 ligands during anthrax infection of macrophages . Inhalation of Bacillus anthracis , a bioterrorism agent , results in a high mortality rate despite appropriate antibiotic therapy . Macrophages appear to be a key factor in B. anthracis pathogenesis . The burst of pro-inflammatory cytokines from macrophages could be a major cause of death in anthrax . However , preactivation of Toll-like receptors ( TLRs ) could modify the host response . TLR ligands stimulate the release of activating cytokines but may also down-modulate the subsequent deleterious cytokine response to pathogens . We developed a cell culture model to measure macrophage responses to B. anthracis spores and bacilli . We found that germination from spores to bacilli produced a substantial stimulus for the secretion of the cytokines P05231 , P01375 , P22301 , and IL-12 p40 . Our studies showed that pretreatment of mouse macrophages with the Q9NR96 ligand DB05463 , or the Q9NYK1 ligands R-848 and IT-37 , results in a substantial decrease in the subsequent secretion of P05231 and P01375 in response to B. anthracis infection of macrophages . Furthermore , the Q9NYK1 and Q9NR96 ligands significantly decreased anthrax-induced cytotoxicity in the macrophages . These findings suggest that TLR ligands may contribute to the enhancement of innate immunity in B. anthracis infection by suppressing potentially deleterious pro-inflammatory cytokine responses and by improving macrophage viability . Candidate gene studies of ADHD : a meta-analytic review . Quantitative genetic studies ( i.e. , twin and adoption studies ) suggest that genetic influences contribute substantially to the development of attention deficit hyperactivity disorder ( ADHD ) . Over the past 15 years , considerable efforts have been made to identify genes involved in the etiology of this disorder resulting in a large and often conflicting literature of candidate gene associations for ADHD . The first aim of the present study was to conduct a comprehensive meta-analytic review of this literature to determine which candidate genes show consistent evidence of association with childhood ADHD across studies . The second aim was to test for heterogeneity across studies in the effect sizes for each candidate gene as its presence might suggest moderating variables that could explain inconsistent results . Significant associations were identified for several candidate genes including Q01959 , P21917 , P21918 , P31645 , P28222 , and P60880 . Further , significant heterogeneity was observed for the associations between ADHD and Q01959 , P21917 , P21918 , P09172 , P08913 , P31645 , Q8IWU9 , P21397 , and P60880 , suggesting that future studies should explore potential moderators of these associations ( e.g. , ADHD subtype diagnoses , gender , exposure to environmental risk factors ) . We conclude with a discussion of these findings in relation to emerging themes relevant to future studies of the genetics of ADHD . Effects of ozone exposure mediated by BEAS-2B cells on T cells activation : a possible link between environment and asthma . OBJECTIVE : To explore the possible link between ozone and asthma through analyzing Th1/Th2 differentiation of T cells following incubation with conditioned medium from the BEAS-2B cells exposed to ozone in vitro . METHOD : Bronchial epithelial cell line , BEAS-2B , was cultured using an air-liquid interface culture system in a CO2 incubator and exposed to 0 or 0.16 or 0.25 mg/m3 of ozone for 8 h . The amounts of IL-1β , P05231 and RANTES in the cell supernatant were detected . The cell culture supernatants were collected and used as conditioned medium in the next experiment . T cells from children recruited were incubated with conditioned medium for 12 h . Activation rate of Q07108 and Th1/Th2/Th17 differentiation were analyzed . RESULTS : BEAS-2B cells exposed to different ozone concentrations showed morphological changes . Cells exposed to 0.16 and 0.25 mg/m3 ozone produced higher amounts of IL-1β , P05231 and RANTES than that in the control group . Children with allergic asthma had upregulated expression of genes related with asthma , including P13500 , CCR4 , P19875 , Q9Y271 , Q99665 , Q14627 , Q13478 , P01584 , P10145 , P25025 and O75888 . Q07108 expression in T cells was significantly elevated irrespective of ozone exposure in children with allergic asthma . Following ozone exposure , in asthmatic children group , expression levels of cytokines of Th1 cells were collectively higher than those from Th2 cells . Ozone-exposed conditioned media could slightly increase all the Th1 , Th2 and Th17 cytokines in T cells from allergic asthmatic children . CONCLUSIONS : Our results suggested that Th1 cells activation might be predominant over Th2 activation upon ozone exposure in asthmatic children , which might help to clarify the mechanisms of asthma related to environmental factors like ozone . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . Nicotinic acetylcholine receptors containing the α6 subunit contribute to ethanol activation of ventral tegmental area dopaminergic neurons . DB00184 and alcohol are often co-abused suggesting a common mechanism of action may underlie their reinforcing properties . Both drugs acutely increase activity of ventral tegmental area ( VTA ) dopaminergic ( DAergic ) neurons , a phenomenon associated with reward behavior . Recent evidence indicates that nicotinic acetylcholine receptors ( nAChRs ) , ligand-gated cation channels activated by ACh and nicotine , may contribute to ethanol-mediated activation of VTA DAergic neurons although the nAChR subtype(s) involved has not been fully elucidated . Here we show that expression and activation of nAChRs containing the α6 subunit contribute to ethanol-induced activation of VTA DAergic neurons . In wild-type ( WT ) mouse midbrain sections that contain the VTA , ethanol ( 50 or 100 mM ) significantly increased firing frequency of DAergic neurons . In contrast , ethanol did not significantly increase activity of VTA DAergic neurons in mice that do not express Q15825 , the gene encoding the α6 nAChR subunit ( α6 knock-out ( KO ) mice ) . DB00898 -induced activity in WT slices was also reduced by pre-application of the α6 subtype-selective nAChR antagonist , α-conotoxin MII[E11A] . When co-applied , ethanol potentiated the response to ACh in WT DAergic neurons ; whereas co-application of ACh and ethanol failed to significantly increase activity of DAergic neurons in α6 KO slices . Finally , pre-application of α-conotoxin MII[E11A] in WT slices reduced ethanol potentiation of ACh responses . Together our data indicate that α6-subunit containing nAChRs may contribute to ethanol activation of VTA DAergic neurons . These receptors are predominantly expressed in DAergic neurons and known to be critical for nicotine reinforcement , providing a potential common therapeutic molecular target to reduce nicotine and alcohol co-abuse . Angiogenesis , cell proliferation and apoptosis in gastric ulcer healing . Effect of a selective cox-2 inhibitor . To elucidate the role of cyclooxygenase-2 , we compared the effects of rofecoxib , a selective cyclooxygenase-2 inhibitor , and ibuprofen , a nonselective cyclooxygenase inhibitor , on the evolution of acetic-acid-induced gastric ulcers in rats , evaluating growth factor expression , the angiogenic process , cell proliferation and cell apoptosis . Levels of basic fibroblast growth factor ( P09038 ) and vascular endothelial growth factor ( P15692 ) , angiogenesis and cell proliferation were analysed by immunohistochemical methods , and apoptosis was evaluated by an enzyme immunoassay . Both growth factors and microvessels appeared to be abundant in the granulation tissue of the ulcer bed . DB00533 ( 2.5 mg/kg/day ) and ibuprofen ( 100 mg/kg/day ) delayed ulcer healing , but only rofecoxib treatment provoked a reduction of P09038 expression and inhibition of the development of new microvessels . No changes in P15692 expression were detected . Results also showed that proliferation and apoptosis were increased in control ulcerated animals . DB00533 reduced significantly both processes . These findings demonstrate that a reduction of P09038 expression and an antiangiogenic action , as well as proliferation/apoptosis inhibition , are some of the mechanisms possibly implicated in the delay in ulcer healing seen after the administration of the highly selective P35354 inhibitor rofecoxib . Molecular genetics of attention deficit hyperactivity disorder . Although twin studies demonstrate that ADHD is a highly heritable condition , molecular genetic studies suggest that the genetic architecture of ADHD is complex . The handful of genome-wide linkage and association scans that have been conducted thus far show divergent findings and are , therefore , not conclusive . Similarly , many of the candidate genes reviewed here ( ie , P09172 , P21397 , P23975 , P17752 -2 , P31645 , P43681 , Q12879 ) are theoretically compelling from neurobiological systems perspective but available data are sparse and inconsistent . However , candidate gene studies of ADHD have produced substantial evidence implicating several genes in the etiology of the disorder , with meta-analyses supportive of a role of the genes coding for P21917 , P21918 , Q01959 , P60880 , and P28222 in the etiology of ADHD . Pharmacological investigation of the role of leukotrienes in the pathogenesis of experimental NSAID gastropathy . The role of leukotrienes in the pathogenesis of acute gastric ulceration induced by nonsteroidal antiinflammatory drugs was investigated using a rat model . One part of the study involved oral pretreatment with a leukotriene synthesis inhibitor 1 h prior to administration of indomethacin ( 20 mg/kg per os ) . Three hours after indomethacin , the extent of macroscopically visible gastric damage was determined , and gastric LTB4 synthesis was determined . The compounds tested were PF-5901 , A-64077 , nordihydroguaiaretic acid , and L-698,037 . Each compound produced dose-related inhibition of gastric LTB4 synthesis and a parallel reduction in the severity of indomethacin-induced damage . The antioxidant properties of these compounds was assessed using an in vitro assay . There was no correlation between the antioxidant properties of the compounds and their ability to reduce the severity of indomethacin-induced gastric damage . In the second part of the study , the effects of intravenous , administration of LTD4 and LTB4 receptor antagonists on indomethacin-induced gastric epithelial damage ( measured by permeability to [51Cr] DB00974 ) were assessed . The two Q9Y271 antagonists ( MK-571 and DB00549 ) significantly reduced the permeability changes induced by indomethacin , while the two LTB4 antagonists ( SC-41930 and LY-255,283 ) were without significant effect . Despite the reduction of gastric epithelial injury , blockade of LTD4 receptors did not markedly affect the extent of macroscopically visible injury . These data are consistent with the hypothesis that leukotrienes contribute to the epithelial injury and macroscopically visible damage induced by NSAIDs . However , it remains unclear to what extent leukotrienes are involved in the initiation of the injury , as opposed to its amplification . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Genotype frequencies of 50 polymorphisms for 241 Japanese non-cancer patients . This paper lists the genotype frequencies of 50 polymorphisms of 37 genes ( P05091 , P07550 , P13945 , P21964 , P16671 , P25025 , P24385 , P35354 , P11509 , P05093 , P11511 , IGF1 , IL-1A , IL-1B , IL-1RN , IL-1R1 , P05231 , P10145 , P22301 , P41159 , Le , L-myc , P05164 , Q99707 , P42898 , P21397 , P15559 , O15527 , p53 , p73 , Se , P31213 , TGF-B , P01375 -A , P01375 -B , P18074 , and P18887 ) and 6 sets of combined genotype frequencies for 241 non-cancer Japanese outpatients . Though the genotype frequencies of 25 polymorphisms have already been reported in our previous papers , 15 polymorphisms ( P16671 A52C , P25025 C785T , P24385 G870A , IGF1 C/T at intron 2 and G2502T , IL-1A 46-bp VNTR , IL-1R1 C-116T , P05231 Ins/Del 17C , P10145 A-278T and C74T , IL- 10 T-819C , P41159 A-2548G , P31213 2-bp VNTR , P18074 Lys751Gln , and P18887 Arg399Gln ) and six sets of combined genotype frequencies ( IL-1B C-31T and IL-1A C-889T , IL-1B C-31T and IL-1RN 86-bp VNTR , IL-1B C-31T and IL-1R1 C-116T , P01375 -A G-308A and P01375 -B A252G , P31213 Val89Leu and 2-bp VNTR , and P18887 Arg399Gln and P18074 Lys751Gln ) were reported in this paper for the first time for Japanese . Although microarray technology will produce this kind of information in near future , this is the first document that reports the genotype/allele frequencies among Japanese for an archival purpose . Effect of nimesulide on proliferation and apoptosis of human hepatoma SMMC-7721 cells . AIM : P35354 ( P35354 ) has been suggested to be associated with carcinogenesis . We sought to investigate the effect of the selective P35354 inhibitor , DB04743 on proliferation and apoptosis of SMMC-7721 human hepatoma cells . METHODS : This study was carried out on the culture of hepatic carcinoma SMMC-7721 cell line . Various concentrations of DB04743 ( 0 , 200 micromol/L , 300 micromol/L , 400 micromol/L ) were added and incubated . Cell proliferation was detected with MTT colorimetric assay , cell apoptosis by electron microscopy , flow cytometry and TUNEL . RESULTS : DB04743 could significantly inhibit SMMC-7721 cells proliferation dose-dependent and in a dependent manner compared with that of the control group . The duration lowest inhibition rate produced by DB04743 in SMMC-7721 cells was 19.06 % , the highest inhibition rate was 58.49 % . After incubation with DB04743 for 72 h , the most highest apoptosis rate and apoptosis index of SMMC-7721 cells comparing with those of the control were 21.20 % +/-1.62 % vs 2.24 % +/-0.26 % and 21.23+/-1.78 vs 2.01+/-0.23 ( P < 0.05 ) . CONCLUSION : The selective P35354 inhibitor , DB04743 can inhibit the proliferation of SMMC-7721 cells and increase apoptosis rate and apoptosis index of SMMC-7721 cells . The apoptosis rate and the apoptosis index are dose-dependent . Under electron microscope SMMC-7721 cells incubated with 300 micromol and 400 micromol DB04743 show apoptotic characteristics . With the clarification of the mechanism of selective P35354 inhibitors , These P35354 selective inhibitors can become the choice of prevention and treatment of cancers . Prediction of genetic risk for dyslipidemia . The purpose of the present study was to identify genetic variants that confer susceptibility to dyslipidemia . A total of 5213 individuals from two independent populations were examined : Subject panel A comprised 3794 individuals who visited participating hospitals ; subject panel B comprised 1419 community-dwelling elderly individuals . The genotypes for 100 polymorphisms of 65 candidate genes were determined . The chi(2) test and multivariable logistic regression analysis revealed that seven polymorphisms of Q6Q788 , P02656 , P02647 , ACAT2 , and P06858 were significantly associated with hypertriglyceridemia , six polymorphisms of Q6Q788 , P11150 , and P08684 with low HDL-cholesterol , and three polymorphisms of P02649 and P41597 with high LDL-cholesterol in subject panel A . For validation of these associations , the same polymorphisms were examined in subject panel B . Six polymorphisms of Q6Q788 , P02656 , P02647 , and P06858 were again significantly associated with hypertriglyceridemia , three polymorphisms of Q6Q788 with low HDL-cholesterol , and two polymorphisms of P02649 with high LDL-cholesterol . Serum triglyceride , HDL-cholesterol , and LDL-cholesterol concentrations differed significantly among genotypes of these corresponding polymorphisms in both subject panels . These results indicate that polymorphisms of Q6Q788 , P02656 , P02647 , and P06858 are determinants of hypertriglyceridemia and that those of Q6Q788 and P02649 are determinants of low HDL-cholesterol and high LDL-cholesterol , respectively , in Japanese individuals .

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interacts_with DB05039?

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Decreasing Poly(ADP- DB01936 ) Polymerase Activity Restores ΔF508 P13569 Trafficking . Most cystic fibrosis is caused by mutations in P13569 that prevent its trafficking from the ER to the plasma membrane and is associated with exaggerated inflammation , altered metabolism , and diminished responses to oxidative stress . P09874 is activated by oxidative stress and causes energy depletion and cell dysfunction . Inhibition of this enzyme protects against excessive inflammation and recent studies have also implicated it in intracellular protein trafficking . We hypothesized that P09874 activity is altered in CF and affects trafficking and function of the most common CF mutant ΔF508 P13569 . Indeed , P09874 activity was 2.9-fold higher in CF ( ΔF508/ΔF508 ) human bronchial epithelial primary cells than in non-CF cells , and similar results were obtained by comparing CF vs. non-CF bronchial epithelial cell lines ( 2.5-fold higher in CFBE41o(-) vs. 16HBE14o(-) , P < 0.002 ) . A P09874 inhibitor ( ABT-888 , DB07232 ) partially restored P13569 channel activity in CFBE41o(-) cells overexpressing ΔF508 P13569 . Similarly , reducing P09874 activity by 85 % in ileum from transgenic CF mice ( Cftr(tm1)Eur ) partially rescued ΔF508 P13569 activity to 7 % of wild type mouse levels , and similar correction ( 7.8 % ) was observed in vivo by measuring salivary secretion . Inhibiting P09874 with ABT-888 or siRNA partially restored ΔF508 P13569 trafficking in cell lines , and most ΔF508 P13569 was complex glycosylated when heterologously expressed in P09874 (-/-) mouse embryonic fibroblasts . Finally , levels of the mature glycoform of P13569 were reduced by peroxynitrite , a strong activator of P09874 . These results demonstrate that P09874 activity is increased in CF , and identify a novel pathway that could be targeted by proteostatic correctors of P13569 trafficking . A screen for peptide agonists of the Q99062 . BACKGROUND : Granulocyte-colony stimulating factor ( DB00099 ) is one of the most important pharmacologically used proteins . Potential uses beyond the stimulation of neutrophilic granulocytes are the treatment of CNS disorders . Disadvantages of the G- P04141 protein as a drug are its moderate plasma half-life time and considerable production costs . We therefore conducted a screen for peptide agonists derived from the sequence of human DB00099 . FINDINGS : Despite of the high sensitivity of our screening system we could not detect any positive hits in a single peptide approach . In a multiplex approach using a permutation of any combination of 10 different peptides we could also not detect a positive block . CONCLUSIONS : We conclude that larger coherent parts of the protein or dimerising peptides may be needed to achieve activation of the receptor . Distinctive RNA expression profiles in blood associated with Alzheimer disease after accounting for white matter hyperintensities . BACKGROUND : Defining the RNA transcriptome in Alzheimer Disease ( AD ) will help understand the disease mechanisms and provide biomarkers . Though the AD blood transcriptome has been studied , effects of white matter hyperintensities ( WMH ) were not considered . This study investigated the AD blood transcriptome and accounted for WMH . METHODS : RNA from whole blood was processed on whole-genome microarrays . RESULTS : A total of 293 probe sets were differentially expressed in AD versus controls , 5 of which were significant for WMH status . The 288 AD-specific probe sets classified subjects with 87.5 % sensitivity and 90.5 % specificity . They represented 188 genes of which 29 have been reported in prior AD blood and 89 in AD brain studies . Regulated blood genes included P14780 , MME ( P08473 ) , TGFβ1 , P22748 , Q16625 , Q13315 , Q08188 , IGFR2 , NOV , Q63HN8 , P51813 , LRRN1 , Q13555 , P06213 , P07339 , Q8WY21 , Q92673 , and Q9HCD6 . CONCLUSIONS : RNA expression is altered in AD blood irrespective of WMH status . Some genes are shared with AD brain . Surfactin exhibits neuroprotective effects by inhibiting amyloid β-mediated microglial activation . Microglial-mediated neuroinflammation and neurotoxicity contribute to the pathogenesis of neurodegenerative diseases including Alzheimer 's disease ; therefore , control of microglial activation and subsequent suppression of neurotoxic pro-inflammatory molecules could provide a potential therapeutic approach for the treatment of such diseases . In this study , we investigated the effects of surfactin , a surfactant from Bacillus subtilis , on oligomeric amyloid β ( Aβ ) -induced microglial activation and neurotoxicity . Surfactin significantly suppressed expression of P14780 , P35228 and P35354 , as well as production of ROS , NO , DB00917 , P01375 -α , IL-1β , P05231 and P13500 in Aβ-stimulated BV-2 microglial cells . Moreover , surfactin markedly inhibited Aβ-induced nuclear translocation and activation of NF-κB as well as phosphorylation of JNK and p38 MAPK . Furthermore , surfactin protected hippocampal HT22 cells from indirect neuronal toxicity mediated by Aβ-treated microglial cells , but had no effect on Aβ-induced direct toxicity to HT22 cells . These results suggest that surfactin impairs the Aβ-induced inflammatory response of microglial cells and confers protection against indirect neurotoxicity to hippocampal cells . Our findings indicate that surfactin may have therapeutic potential for ameliorating Alzheimer 's disease as well as other neurodegenerative disorders which involve neuroinflammation . The hematopoietic factor granulocyte-colony stimulating factor improves outcome in experimental spinal cord injury . Granulocyte-colony stimulating factor ( DB00099 ) is a potent hematopoietic factor that drives differentiation of neutrophilic granulocytes . We have recently shown that G- P04141 also acts as a neuronal growth factor , protects neurons in vitro and in vivo , and has regenerative potential in various neurological disease models . Spinal cord injury ( SCI ) following trauma or secondary to skeletal instability is a terrible condition with no effective therapies available at present . In this study , we show that the Q99062 is up-regulated upon experimental SCI and that G- P04141 improves functional outcome in a partial dissection model of SCI . G- P04141 significantly decreases apoptosis in an experimental partial spinal transsection model in the mouse and increases expression of the anti-apoptotic G- P04141 target gene Bcl-X(L) . In vitro , G- P04141 enhances neurite outgrowth and branching capacity of hippocampal neurons . In vivo , G- P04141 treatment results in improved functional connectivity of the injured spinal cord as measured by Mn(2+)-enhanced Q9BWK5 . G- P04141 also increased length of the dorsal corticospinal tract and density of serotonergic fibers cranial to the lesion center . Mice treated systemically with G- P04141 as well as transgenic mice over-expressing G- P04141 in the CNS exhibit a strong improvement in functional outcome as measured by the BBB score and gridwalk analysis . We show that G- P04141 improves outcome after experimental SCI by counteracting apoptosis , and enhancing connectivity in the injured spinal cord . We conclude that G- P04141 constitutes a promising and feasible new therapy option for SCI . No effect of the oral neutral endopeptidase inhibitor candoxatril , on bronchomotor tone and histamine reactivity in asthma . P08473 ( NEP ) is found in many tissues in man , including the lung . Metabolism by NEP is one of the main mechanisms for the clearance of atrial natriuretic peptide ( P01160 ) , a hormone that causes bronchodilation and reduces nonspecific bronchial reactivity in man . DB00616 , an oral NEP inhibitor has been shown to elevate circulating P01160 levels . We have sought to determine whether the administration of candoxatril will alter bronchomotor tone ( forced expiratory volume in one second ( FEV1 ) ) and histamine reactivity . Ten male asthmatic patients with stable asthma were enrolled ( mean ( SD ) age 32 ( 10 ) yrs ; FEV1 92 ( 11 ) % predicted ) in a randomized , double-blind , placebo-controlled study . On each study day , after baseline spirometry , patients received 200 mg of candoxatril or placebo . Spirometry was repeated at half hourly intervals . After 2 h a histamine inhalation test was performed . There was no significant difference in FEV1 values at baseline or at 2 h post-dosing between active and placebo study days , with mean ( SEM ) FEV1 at baseline and 2 h of 3.71 ( 0.29 ) l and 3.85 ( 0.29 ) l on the placebo day , and 3.89 ( 0.27 ) l and 4.05 ( 0.82 ) l on the active day , respectively . The geometric mean ( range ) provocative concentration of histamine producing a 20 % fall in FEV1 ( PC20 ) on the placebo day and active day did not differ significantly , being 1.17 ( 0.25-25.8 ) and 0.93 ( 0.13-32 ) mg.ml-1 , respectively. ( ABSTRACT TRUNCATED AT 250 WORDS ) Antitumor activity of Ru(III) complexes carrying beta-diketonato ligands in vitro and in vivo . PURPOSE : To investigate the antitumor activity of two newly synthesized ruthenium(III) [ Ru(III) ] compounds carrying bidentate ligands : (acac)-acetylacetonate , [ Ru(acac)3 ) , and (tfac)-trifluoroacetylacetonate [ Ru(tfac)3 ] . MATERIALS AND METHODS : The activity of ruthenium(III) analogues was evaluated on HeLa , B16 , and Femx cell lines for cytotoxicity in vitro using MTT assay , and inhibition on tumor invading ability in vitro using cell migration and invasion assays , whereas inhibition of tumor growth in vivo was estimated on advanced B16 murine melanoma model . Both compounds were also investigated in combinations with cisplatin , oxaliplatin , or poly ADP-ribose polymerase- 1 ( P09874 ) inhibitor , in order to determine the pattern of mutual interactions . RESULTS : Applied as single drugs , Ru(tfac)3 showed high cytotoxic activity against HeLa and Femx cell lines , while Ru(acac)3 did not reach the IC50 on any of the cell lines tested . In combinations , Ru(acac)3 with cisplatin gained synergistic interaction , antagonistic with oxaliplatin , and of different kind with ( P09874 ) inhibitor in concentration-and cell line-dependent manner . Ru(acac)3 exhibited inhibition of HeLa cell migration and gelatinolytic activity of P08253 and P14780 . Ru(tfac)3 complexes did not induce significant reduction of melanoma growth in vivo , whereas Ru(acac)3 did , but the latter failed to contribute in lifespan improvement . CONCLUSION : The investigated ruthenium complexes showed different levels of antitumor activity in vitro and in vivo , implicating on different mechanisms of their action as well as diverse perspectives in cancer treatment . DB01427 suppresses the synthesis of tumor necrosis factor-alpha in human mononuclear cells . P01375 -alpha ( P01375 ) exerts a wide spectrum of biological activities and contributes to the pathophysiology of septic shock . Elevated circulating levels of P01375 have also been reported in patients with severe chronic heart failure . We studied the effect of amrinone , a class III cyclic nucleotide phosphodiesterase inhibitor used in the treatment of acute heart failure , on the synthesis of P01375 in vitro . Peripheral blood mononuclear cells from healthy volunteers or cells of a permanent monoblast cell line were stimulated for 20 h with bacterial lipopolysaccharide and different doses of amrinone . P01375 production is suppressed in a dose-dependent manner to a minimum of 9 % of controls with 1000 microM of amrinone , reaching half-maximal inhibition at 80 microM amrinone . This effect appears to be mediated via DB02527 , which accumulated nearly twofold in the presence of amrinone . Suppression of P01375 synthesis by therapeutically administered phosphodiesterase inhibitors such as amrinone may contribute to their beneficial effect in the treatment of heart failure . Characterization of de novo synthesized GPCRs supported in nanolipoprotein discs . The protein family known as G-protein coupled receptors ( GPCRs ) comprises an important class of membrane-associated proteins , which remains a difficult family of proteins to characterize because their function requires a native-like lipid membrane environment . This paper focuses on applying a single step method leading to the formation of nanolipoprotein particles ( NLPs ) capable of solubilizing functional GPCRs for biophysical characterization . NLPs were used to demonstrate increased solubility for multiple GPCRs such as the Neurokinin 1 Receptor ( P25103 ) , the Adrenergic Receptor â2 ( P07550 ) and the Dopamine Receptor D1 ( P21728 ) . All three GPCRs showed affinity for their specific ligands using a simple dot blot assay . The P25103 was characterized in greater detail to demonstrate correct folding of the ligand pocket with nanomolar specificity . Electron paramagnetic resonance ( EPR ) spectroscopy validated the correct folding of the P25103 binding pocket for Substance P ( SP ) . Fluorescence correlation spectroscopy ( FCS ) was used to identify SP-bound P25103 -containing NLPs and measure their dissociation rate in an aqueous environment . The dissociation constant was found to be 83 nM and was consistent with dot blot assays . This study represents a unique combinational approach involving the single step de novo production of a functional GPCR combined with biophysical techniques to demonstrate receptor association with the NLPs and binding affinity to specific ligands . Such a combined approach provides a novel path forward to screen and characterize GPCRs for drug discovery as well as structural studies outside of the complex cellular environment . Targeting the epigenome in the treatment of asthma and chronic obstructive pulmonary disease . Epigenetic modification of gene expression by methylation of DNA and various post-translational modifications of histones may affect the expression of multiple inflammatory genes . Acetylation of histones by histone acetyltransferases activates inflammatory genes , whereas histone deacetylation results in inflammatory gene repression . Corticosteroids exert their antiinflammatory effects partly by inducing acetylation of antiinflammatory genes , but mainly by recruiting histone deacetylase-2 ( Q92769 ) to activated inflammatory genes . Q92769 deacetylates acetylated glucocorticoid receptors so that they can suppress activated inflammatory genes in asthma . In chronic obstructive pulmonary disease ( P48444 ) , there is resistance to the antiinflammatory actions of corticosteroids , which is explained by reduced activity and expression of Q92769 . This can be reversed by a plasmid vector , which restores Q92769 levels , but may also be achieved by low concentrations of theophylline . Oxidative stress causes corticosteroid resistance by reducing Q92769 activity and expression by activation of phosphoinositide-3-kinase-delta , resulting in Q92769 phosphorylation via a cascade of kinases . DB00277 reverses corticosteroid resistance by directly inhibiting oxidant-activated O00329 and is mimicked by O00329 knockout or by selective inhibitors . Other treatments may also interact in this pathway , making it possible to reverse corticosteroid resistance in patients with P48444 , as well as in smokers with asthma and some patients with severe asthma in whom similar mechanisms operate . Other histone modifications , including methylation , tyrosine nitration , and ubiquitination may also affect histone function and inflammatory gene expression , and better understanding of these epigenetic pathways could led to novel antiinflammatory therapies , particularly in corticosteroid-resistant inflammation . DB05039 inhibits tumor cell invasiveness and P14780 expression by suppressing IKK/NF-κB activation . The β2 adrenergic receptor ( P07550 ) is a G protein-coupled transmembrane receptor expressed in the human respiratory tract and widely recognized as a pharmacological target for treatments of asthma and chronic obstructive pulmonary disorder ( P48444 ) . Although a number of P07550 agonists have been developed for use in asthma therapy , indacaterol is the only ultra-long-acting inhaled β2-agonist ( LABA ) approved by the FDA for relieving the symptoms in P48444 patients . The precise molecular mechanism underlying the pharmacological effect of indacaterol , however , remains unclear . Here , we show that β-arrestin-2 mediates the internalization of P07550 following indacaterol treatment . Moreover , we demonstrate that indacaterol significantly inhibits tumor necrosis factor-α ( P01375 -α ) -induced NF-κB activity by reducing levels of both phosphorylated-IKK and -IκBα , thereby decreasing NF-κB nuclear translocation and the expression of P14780 , an NF-κB target gene . Subsequently , we show that indacaterol significantly inhibits P01375 -α/NF-κB-induced cell invasiveness and migration in a human cancer cell line . In conclusion , we propose that indacaterol may inhibit NF-κB activity in a β-arrestin2-dependent manner , preventing further lung damage and improving lung function in P48444 patients . Supplementation of Lactobacillus curvatus HY7601 and Lactobacillus plantarum KY1032 in diet-induced obese mice is associated with gut microbial changes and reduction in obesity . OBJECTIVE : To investigate the functional effects of probiotic treatment on the gut microbiota , as well as liver and adipose gene expression in diet-induced obese mice . DESIGN : Male C57BL/6J mice were fed a high-fat diet ( HFD ) for 8 weeks to induce obesity , and then randomized to receive HFD+probiotic ( Lactobacillus curvatus HY7601 and Lactobacillus plantarum KY1032 , n = 9 ) or HFD+placebo ( n = 9 ) for another 10 weeks . Normal diet ( ND ) fed mice ( n = 9 ) served as non-obese controls . RESULTS : Diet-induced obese mice treated with probiotics showed reduced body weight gain and fat accumulation as well as lowered plasma insulin , leptin , total-cholesterol and liver toxicity biomarkers . A total of 151,061 pyrosequencing reads for fecal microbiota were analyzed with a mean of 6,564 , 5,274 and 4,464 reads for the ND , HFD+placebo and HFD+probiotic groups , respectively . Gut microbiota species were shared among the experimental groups despite the different diets and treatments . The diversity of the gut microbiota and its composition were significantly altered in the diet-induced obese mice and after probiotic treatment . We observed concurrent transcriptional changes in adipose tissue and the liver . In adipose tissue , pro-inflammatory genes ( TNFα , P05231 , IL1β and MCP1 ) were down-regulated in mice receiving probiotic treatment . In the liver , fatty acid oxidation-related genes ( PGC1α , CPT1 , P23786 and Q15067 ) were up-regulated in mice receiving probiotic treatment . CONCLUSIONS : The gut microbiota of diet-induced obese mice appears to be modulated in mice receiving probiotic treatment . Probiotic treatment might reduce diet-induced obesity and modulate genes associated with metabolism and inflammation in the liver and adipose tissue . Transient estrogen exposure from birth affects uterine expression of developmental markers in neonatal gilts with lasting consequences in pregnant adults . Disruption of estrogen-sensitive , estrogen receptor ( ER ) -dependent events during porcine uterine development between birth ( postnatal day= P01160 0 ) and P01160 14 affects patterns of uterine morphoregulatory gene expression in the neonate with lasting consequences for reproductive success . Uterine capacity for conceptus support is reduced in pregnant adult gilts exposed to estradiol valerate ( EV ) for 14 days from birth . Objectives here were to determine effects of EV exposure from birth through P01160 13 on neonatal uterine and adult endometrial markers of growth , patterning , and remodeling . Targets included the relaxin receptor ( Q9HBX9 ) , estrogen receptor-alpha ( P03372 ) and vascular endothelial growth factor ( P15692 ) , morphoregulatory markers P31260 and O00755 , and the matrix metalloproteinases (MMP)2 and P14780 . Gilts were treated daily with EV ( 50 microg/kg body weight per day , i.m. ) or corn oil vehicle from birth through P01160 13 . Uteri were obtained from neonates on P01160 14 and from adults on pregnancy day 12 ( PxD 12 ) . In neonates , EV exposure from birth increased uterine Q9HBX9 gene expression , and both P03372 and P15692 proteins . At PxD 12 , endometrial Q9HBX9 mRNA remained elevated , while P03372 protein was reduced . Early EV treatment decreased neonatal uterine O00755 , but increased P31260 expression . O00755 expression was reduced in EV-treated adults . Transient EV exposure increased P14780 transcripts at P01160 14 , whereas both latent and active P14780 activity was increased due to early EV treatment in adults on PxD 12 . Results support the hypothesis that transient , estrogen-induced disruption of porcine uterine development from birth alters early programming events that lead to functional consequences in the adult . Different mechanisms for gamma-glutamyltransferase-dependent resistance to carboplatin and cisplatin . In this work , we investigated the effect of gamma-glutamyltransferase ( P19440 ) overexpression on cell viability after carboplatin treatment and compared with cisplatin . Carboplatin challenge of HeLa cells induced P19440 and glutamate-cystine ligase ( GCL ) activities by 2- and 1.4-fold , respectively and concomitantly increased the intracellular reduced glutathione ( DB00143 ) level ( 1.5-fold ) . To study the role of P19440 , HeLa- P19440 cells , a stably transfected cell line overexpressing P19440 ( 120-150 mU/mg protein ) and the parental HeLa cells ( 10-15 mU/mg protein ) were used . Both cell lines exhibited comparable viability ( IC(50) approximately 150 microM ) after carboplatin treatment when cultured in standard ( 250 microM cystine ) medium . Culture in low ( 50 microM ) cystine medium resulted in a dramatic decrease ( approximately 90 % ) of the intracellular DB00143 level and to a 2.5-fold increase of carboplatin cytotoxicity ( IC(50) approximately 60 microM ) . When DB00143 ( 50 microM ) was included in the culture medium , only HeLa- P19440 cells exhibited increased resistance to carboplatin . Using partially purified P19440 from HeLa- P19440 cells , we show that cisplatin forms adducts with cysteinylglycine , depending only on P19440 activity whereas carboplatin did not efficiently react with cysteinylglycine . Thus , in this model system , P19440 activity can affect platinum drugs cytotoxocity by two different ways : cisplatin can be detoxified extracellularly after reaction with the -SH group of cysteinylglycine ; in the case of carboplatin , the supply of DB00143 precursors , initiated by P19440 , increases the intracellular level of the tripeptide and provides enhanced defensive mechanisms to the cell . P14780 leads to claudin-5 degradation via the NF-κB pathway in BALB/c mice with eosinophilic meningoencephalitis caused by Angiostrongylus cantonensis . The epithelial barrier regulates the movement of ions , macromolecules , immune cells and pathogens . The objective of this study was to investigate the role of the matrix metalloproteinase ( MMP ) -9 in the degradation of tight junction protein during infection with rat nematode lungworm Angiostrongylus cantonensis . The results showed that phosphorylation of IκB and NF-κB was increased in mice with eosinophilic meningoencephalitis . Treatment with MG132 reduced the phosphorylation of NF-κB and the activity of P14780 , indicating upregulation of P14780 through the NF-κB signaling pathway . O00501 was reduced in the brain but elevated in the cerebrospinal fluid ( P04141 ) , implying that A. cantonensis infection caused tight junction breakdown and led to claudin-5 release into the P04141 . Degradation of claudin-5 coincided with alteration of the blood- P04141 barrier permeability and treatment with the MMP inhibitor DB02255 attenuated the degradation of claudin-5 . These results suggested that degradation of claudin-5 was caused by P14780 in angiostrongyliasis meningoencephalitis . O00501 could be used for the pathophysiologic evaluation of the blood- P04141 barrier breakdown and tight junction disruption after infection with A. cantonensis . Dietary L-carnitine supplementation increases lipid deposition in the liver and muscle of yellow catfish ( Pelteobagrus fulvidraco ) through changes in lipid metabolism . DB00583 has been reported to improve growth performance and reduce body lipid content in fish . Thus , we hypothesised that carnitine supplementation can improve growth performance and reduce lipid content in the liver and muscle of yellow catfish ( Pelteobagrus fulvidraco ) , a commonly cultured freshwater fish in inland China , and tested this hypothesis in the present study . Diets containing l-carnitine at three different concentrations of 47 mg/kg ( control , without extra carnitine addition ) , 331 mg/kg ( low carnitine ) and 3495 mg/kg ( high carnitine ) diet were fed to yellow catfish for 8 weeks . The low-carnitine diet significantly improved weight gain ( WG ) and reduced the feed conversion ratio ( FCR ) . In contrast , the high-carnitine diet did not affect WG and FCR . Compared with the control diet , the low-carnitine and high-carnitine diets increased lipid and carnitine contents in the liver and muscle . The increased lipid content in the liver could be attributed to the up-regulation of the mRNA levels of SREBP , PPARγ , fatty acid synthase ( FAS ) and ACCa and the increased activities of lipogenic enzymes ( such as FAS , glucose-6-phosphate dehydrogenase , 6-phosphogluconate dehydrogenase and malic enzyme ) and to the down-regulation of the mRNA levels of the lipolytic gene P50416 . The increased lipid content in muscle could be attributed to the down-regulation of the mRNA levels of the lipolytic genes P50416 and Q96AD5 and the increased activity of lipoprotein lipase . In conclusion , in contrast to our hypothesis , dietary carnitine supplementation increased body lipid content in yellow catfish . Effect of the phosphodiesterase III inhibitor amrinone on cytokine and nitric oxide production in immunostimulated J774.1 macrophages . The level of intracellular cyclic nucleotides is a regulatory factor in a variety of immune processes . Increases in intracellular cyclic AMP ( DB02527 ) and/or cyclic GMP ( cGMP ) concentration by the inhibition of phosphodiesterase have been shown to modulate the inflammatory response . DB01427 is a clinically used positive inotropic agent which elevates intracellular DB02527 and cGMP levels by selective inhibition of the phosphodiesterase III isoenzyme . In the current study , we investigated the effect of various concentrations ( 1-300 microM ) of amrinone on lipopolysaccharide-induced production of pro- and anti-inflammatory cytokines and of nitric oxide ( NO ) in vitro . In cultured murine J774.1 macrophages , 1 ng/ml-10 microg/ml of lipopolysaccharide from Escherichia coli O55: P46977 induced production of tumor necrosis factor-alpha ( P01375 ) , interleukin-10 , and nitrite ( breakdown product of NO ) . Pretreatment of cells with amrinone caused a dose-dependent suppression of P01375 production in the concentration range of 1-100 microM . Furthermore , this drug suppressed NO production in the range of 30-300 microM . Similarly to the results in the J774.1 cells , amrinone also inhibited P01375 and NO production in the range of 10-100 microM in primary rat peritoneal macrophages . At 300 microM , but not at lower concentrations , amrinone inhibited interleukin-10 production in lipopolysaccharide-treated J774.1 macrophages . Pretreatment of the macrophages with 100 and 300 microM amrinone increased the lipopolysaccharide-elicited translocation of nuclear factor-kappa B . Taken together , our results indicate that the phosphodiesterase III inhibitor amrinone modulates the activation/production of many pro- and anti-inflammatory factors in endotoxin-stimulated cells . It remains to be further investigated how such immunomodulatory effects contribute to the clinical profile of the agent . Relative contribution of adipose triglyceride lipase and hormone-sensitive lipase to tumor necrosis factor-α ( P01375 -α ) -induced lipolysis in adipocytes . P01375 -α potently stimulates basal lipolysis in adipocytes , which may contribute to hyperlipidemia and peripheral insulin resistance in obesity . Recent studies show that adipose triglyceride lipase ( Q96AD5 ) and hormone-sensitive lipase ( Q05469 ) act sequentially in catalyzing the first two steps of adipose lipolysis in response to β-adrenergic stimulation . Here , we sought to determine their functional roles in P01375 -α-induced lipolysis . Silencing of Q96AD5 expression in adipocytes almost completely abolished basal and P01375 -α-induced glycerol release . In comparison , the glycerol release under the same conditions was only partially decreased upon reduction in expression of either Q05469 or the Q96AD5 coactivator Q8WTS1 . Interestingly , overexpression of Q96AD5 restored the lipolytic rates in cells with silenced Q05469 or Q8WTS1 , indicating a predominant role for Q96AD5 . While expression of Q96AD5 , Q05469 and Q8WTS1 remains mostly unaffected , P01375 -α treatment caused a rapid abrogation of the Q96AD5 inhibitory protein P27469 . P01375 -α drastically decreased the level of P27469 mRNA , and the level of P27469 protein could be maintained by inhibiting proteasomal protein degradation using MG-132 . Furthermore , coexpression of P27469 was able to significantly decrease P01375 -α-stimulated lipolysis mediated by overexpressed Q96AD5 or Q8WTS1 . We propose that the early reduction in P27469 content is permissive for P01375 -α-induced lipolysis . P08473 inhibitor suppresses the early phase of atrial electrical remodeling in a canine rapid atrial pacing model . INTRODUCTION : We examined the acute effects of neutral endopeptidase inhibitor on the hemodynamics and electrical properties of dogs subjected to rapid atrial pacing . METHODS : Ten beagle dogs were used and divided into two groups with and without candoxatril , a neutral endopeptidase inhibitor preadministration . Before and after the 6 hours rapid atrial pacing from the right atrial appendage , the hemodynamics , atrial effective refractory period , and monophasic action potential duration of the right atrial appendage were measured and blood samples were collected . Atrial tissue was also excised after the experiment . RESULTS : DB00616 significantly increased plasma P01160 levels ( Control : 88.4 +/- 50.25 vs. DB00616 : 197.1 +/- 32.09 pg/ml , p = 0.004 ) and prevented reductions in atrial effective refractory period and monophasic action potential duration . We further demonstrated that the treated animals exhibited significantly higher levels of atrial tissue cyclic GMP ( Control : 28.1 +/- 1.60 fmol/mg vs. DB00616 : 44.5 +/- 12.28 fmol/mg , p = 0.034 ) as well as that of plasma cyclic GMP ( Control : 32 +/- 5.5 vs. DB00616 : 42 +/- 7.1 pg/ml , p = 0.028 ) . CONCLUSION : DB00616 suppressed the shortening of atrial effective refractory period and monophasic action potential duration in the rapid atrial pacing model . As plasma P01160 and the atrial tissue levels of cyclic GMP were higher in the DB00616 group than the control , this effect was considered to appear through the reduction of calcium overload caused by P01160 and cyclic GMP . Identification of liver metastasis-related genes in a novel human pancreatic carcinoma cell model by microarray analysis . Pancreatic cancer with liver metastases has a poor prognosis and the molecular mechanisms remain unclear . In this study , SW1990HM , a highly metastatic human pancreatic carcinoma line was subcloned from SW1990 by intrasplenic injection . In vivo and in vitro tumorigenicity , metastatic potential , in vitro invasion , cell growth curves , plate efficiency and S-phase cell numbers were higher in SW1990HM cells . Gene expression profiles of SW1990HM and SW1990 cells showed 40 metastasis-related genes expressed with a 3-fold difference . Thirteen of these 32.5 % ( 13/40 ) were adhesion and extracellular-matrix related and twelve 30 % ( 12/40 ) were cell growth and proliferation related , such as P09238 , P14780 , P09237 , CDH1 , Q09328 , P35221 , IGF1 , P25025 , Q13683 , Q00987 , MET , P30874 and P15692 , which were related to the onset and progression of tumor metastasis . Thus , SW1990HM is an attractive model to study metastasis and identify potential therapeutic targets . S-Adenosylmethionine decarboxylase partially regulates cell growth of HL-60 cells by controlling the intracellular ROS level : Early senescence and sensitization to gamma-radiation . S-Adenosylmethionine decarboxylase ( P17707 ) is a key enzyme for the biosynthesis of spermidine . P17707 -suppressed HL-60 cells overproduced intracellular reactive oxygen species ( ROS ) , which led to cell growth defect and partial cell death . ROS overproduction was caused by a decrease of the total glutathione ( DB00143 ) and the ratio of reduced to oxidized DB00143 , and by an increase of the intracellular iron uptake . When analyzed by real-time polymerase chain reaction , the transcripts of the genes involved in the DB00143 synthesis ( gamma-glutamyl cysteine synthetase , P48637 ) , as well as the gene of the DB00143 -reducing enzyme ( NADP+-dependent isocitrate dehydrogenase ) , were decreased dramatically in these cells . DNA-repairing genes ( Q13315 , PARP , Q06609 and P43246 ) also were not activated transcriptionally . In these situations , excessive ROS induced severe DNA damage , which could not be repaired , and ultimately led the cells to a spontaneous cell death or an early senescence state . For such cells , gamma-radiation and cisplatin , which are direct DNA-damaging agents , were very effective for promoting cell death . How corticosteroids control inflammation : Quintiles Prize Lecture 2005 . Corticosteroids are the most effective anti-inflammatory therapy for many chronic inflammatory diseases , such as asthma but are relatively ineffective in other diseases such as chronic obstructive pulmonary disease ( P48444 ) . Chronic inflammation is characterised by the increased expression of multiple inflammatory genes that are regulated by proinflammatory transcription factors , such as nuclear factor-kappaB and activator protein-1 , that bind to and activate coactivator molecules , which then acetylate core histones to switch on gene transcription . Corticosteroids suppress the multiple inflammatory genes that are activated in chronic inflammatory diseases , such as asthma , mainly by reversing histone acetylation of activated inflammatory genes through binding of liganded glucocorticoid receptors ( GR ) to coactivators and recruitment of histone deacetylase-2 ( Q92769 ) to the activated transcription complex . At higher concentrations of corticosteroids GR homodimers also interact with DNA recognition sites to active transcription of anti-inflammatory genes and to inhibit transcription of several genes linked to corticosteroid side effects . In patients with P48444 and severe asthma and in asthmatic patients who smoke Q92769 is markedly reduced in activity and expression as a result of oxidative/nitrative stress so that inflammation becomes resistant to the anti-inflammatory actions of corticosteroids . DB00277 , by activating HDAC , may reverse this corticosteroid resistance . This research may lead to the development of novel anti-inflammatory approaches to manage severe inflammatory diseases . Disorders of carnitine transport and the carnitine cycle . DB00583 plays an essential role in the transfer of long-chain fatty acids across the inner mitochondrial membrane . This transfer requires enzymes and transporters that accumulate carnitine within the cell ( O76082 carnitine transporter ) , conjugate it with long chain fatty acids ( carnitine palmitoyl transferase 1 , CPT1 ) , transfer the acylcarnitine across the inner plasma membrane ( carnitine-acylcarnitine translocase , O43772 ) , and conjugate the fatty acid back to DB01992 for subsequent beta oxidation ( carnitine palmitoyl transferase 2 , P23786 ) . Deficiency of the O76082 carnitine transporter causes primary carnitine deficiency , characterized by increased losses of carnitine in the urine and decreased carnitine accumulation in tissues . Patients can present with hypoketotic hypoglycemia and hepatic encephalopathy , or with skeletal and cardiac myopathy . This disease responds to carnitine supplementation . Defects in the liver isoform of CPT1 present with recurrent attacks of fasting hypoketotic hypoglycemia . The heart and the muscle , which express a genetically distinct form of CPT1 , are usually unaffected . These patients can have elevated levels of plasma carnitine . O43772 deficiency presents in most cases in the neonatal period with hypoglycemia , hyperammonemia , and cardiomyopathy with arrhythmia leading to cardiac arrest . Plasma carnitine levels are extremely low . Deficiency of P23786 present more frequently in adults with rhabdomyolysis triggered by prolonged exercise . More severe variants of P23786 deficiency present in the neonatal period similarly to O43772 deficiency associated or not with multiple congenital anomalies . Treatment for deficiency of CPT1 , P23786 , and O43772 consists in a low-fat diet supplemented with medium chain triglycerides that can be metabolized by mitochondria independently from carnitine , carnitine supplements , and avoidance of fasting and sustained exercise . Gastroduodenal tolerability of lumiracoxib vs placebo and naproxen : a pilot endoscopic study in healthy male subjects . BACKGROUND : DB01283 ( DB01283 ) is a cyclooxygenase-2 ( P35354 ) selective inhibitor . AIM : To compare the gastroduodenal tolerability of lumiracoxib with placebo and naproxen in a randomized , parallel-group , double-blind study . METHODS : : Sixty-five healthy male subjects were randomized to receive 8 days ' dosing with lumiracoxib 200 mg twice daily ( b.d. ) ( n = 21 ) , placebo ( n = 22 ) or naproxen 500 mg b.d . ( n = 22 ) . Endoscopic evaluations of gastric and duodenal mucosae were conducted at baseline and after 8 days ' dosing . Serum was assayed for ex-vivo concentrations of thromboxane B2 ( TxB2 ) to determine cyclooxygenase-1 ( P23219 ) inhibitory activity . RESULTS : Sixty subjects ( 20 per group ) completed the study . No gastroduodenal erosions were observed in subjects receiving lumiracoxib . Thirteen subjects receiving naproxen developed duodenal erosions . At the gastric site , one subject in each of the naproxen and placebo groups had erosions ; one subject receiving naproxen also developed a small asymptomatic gastric ulcer . Gastrointestinal adverse events accounted for 42.3 % of all adverse events , occurring in 3/21 , 4/22 and 6/22 of the lumiracoxib , placebo and naproxen groups , respectively . TxB2 levels were similar for patients receiving placebo or lumiracoxib , but were reduced by > 95 % in patients receiving naproxen , compared with placebo . CONCLUSIONS : Multiple doses of lumiracoxib resulted in gastroduodenal tolerability similar to placebo and superior to naproxen . DB01283 ( DB01283 ) : a new selective P35354 inhibitor . DB01283 , a new selective P35354 inhibitor , has been recently approved in England and Mexico for the treatment of acute and chronic pain . Although it is the fifth P35354 inhibitor to come to the market , it has a unique structure that could prove to be important in the adverse event profile . Double blind randomised trials have proved its efficacy in acute pain , dysmenorrhea , rheumatoid arthritis and osteoarthritis . Its gastrointestinal safety profile has been studied in multiple trials . The main clinical trail , therapeutic arthritis research and gastrointestinal event trial , has as primary end point : perforations , obstructions and bleeding and as secondary end points : cardiovascular , renal and hepatic safety profile . The results of this trial will probably change the way we look at selective P35354 inhibitors . Analog of somatostatin vapreotide exhibits biological effects in vitro via interaction with neurokinin-1 receptor . OBJECTIVES : DB04894 , a synthetic analog of somatostatin , has analgesic activity most likely mediated through the blockade of neurokinin-1 receptor ( P25103 ) , the DB05875 ( SP ) -preferring receptor . The ability of vapreotide to interfere with other biological effects of SP has yet to be investigated . METHODS : We studied the ability of vapreotide to antagonize P25103 in three different cell types : immortalized U373MG human astrocytoma cells , human monocyte-derived macrophages ( MDM ) and a human embryonic kidney cell line , HEK293 . Both U373MG and MDM express endogenous P25103 while HEK293 cells , which normally do not express P25103 , are stably transformed to express human P25103 ( HEK293- P25103 ) . RESULTS : DB04894 attenuates SP-triggered intracellular calcium increases and nuclear factor-κB activation in a dose-dependent manner . DB04894 also inhibits SP-induced interleukin-8 and monocyte chemotactic protein-1 production in HEK293- P25103 and U373MG cell lines . DB04894 inhibits HIV-1 infection of human MDM in vitro , an effect that is reversible by SP pretreatment . CONCLUSIONS : Our findings indicate that vapreotide has P25103 antagonist activity and may have a potential application as a therapeutic intervention in HIV-1 infection . Enhanced killing of cancer cells by poly(ADP-ribose) polymerase inhibitors and topoisomerase I inhibitors reflects poisoning of both enzymes . Poly(ADP-ribose) polymerase-1 ( P09874 ) plays critical roles in the regulation of DNA repair . Accordingly , small molecule inhibitors of PARP are being developed as agents that could modulate the activity of genotoxic chemotherapy , such as topoisomerase I poisons . In this study we evaluated the ability of the PARP inhibitor veliparib to enhance the cytotoxicity of the topoisomerase I poisons topotecan and camptothecin ( CPT ) . DB07232 increased the cell cycle and cytotoxic effects of topotecan in multiple cell line models . Importantly , this sensitization occurred at veliparib concentrations far below those required to substantially inhibit poly(ADP-ribose) polymer synthesis and at least an order of magnitude lower than those involved in selective killing of homologous recombination-deficient cells . Further studies demonstrated that veliparib enhanced the effects of CPT in wild-type mouse embryonic fibroblasts ( MEFs ) but not Parp1(-/-) MEFs , confirming that P09874 is the critical target for this sensitization . Importantly , parental and Parp1(-/-) MEFs had indistinguishable CPT sensitivities , ruling out models in which P09874 catalytic activity plays a role in protecting cells from topoisomerase I poisons . To the contrary , cells were sensitized to CPT in a veliparib-independent manner upon transfection with P09874 E988K , which lacks catalytic activity , or the isolated P09874 DNA binding domain . These results are consistent with a model in which small molecule inhibitors convert P09874 into a protein that potentiates the effects of topoisomerase I poisons by binding to damaged DNA and preventing its normal repair . Novel synthesis of various orthogonally protected Cα-methyllysine analogues and biological evaluation of a vapreotide analogue containing ( S ) -α-methyllysine . Prochiral malonic diesters containing a quaternary carbon center have been successfully transformed into a diverse set of (t)Boc-Fmoc-α(2,2)-methyllysine-OH analogues through chiral malonic half-ester intermediates obtained via enzymatic ( Pig Liver Esterase , PLE ) hydrolysis . The variety of chiral half-ester intermediates , which vary from 1 to 6 methylene units in the side chain , are achieved in moderate to high optical purity and in good yields . The PLE hydrolysis of malonic diesters with various side chain lengths appears to obey the Jones 's PLE model according to the stereochemical configurations of the resulting chiral half-esters . The established synthetic strategy allows the construction of both enantiomers of α(2,2)-methyllysine analogues , and a ( S ) -β(2,2)-methyllysine analogue from a common synthon by straightforward manipulation of protecting groups . Two different straightforward and cost effective synthetic strategies are described for the synthesis of α(2,2)-methyllysine analogues . The described strategies should find significant usefulness in preparing novel peptide libraries with unnatural lysine analogues . A DB04894 analogue incorporating ( S ) -α(2,2)-methyllysine was prepared . However , the DB04894 analogue with ( S ) -α-methyl-α-lysine is found to lose its specific binding to somatostatin receptor subtype 2 ( P30874 ) . Detecting early kidney damage in horses with colic by measuring matrix metalloproteinase -9 and -2 , other enzymes , urinary glucose and total proteins . BACKGROUND : The aim of the study was to investigate urine matrix metalloproteinase ( P08253 and -9 ) activity , alkaline phosphatase/creatinine ( U-AP/Cr ) and gamma-glutamyl-transpeptidase/creatinine ( U- P19440 /Cr ) ratios , glucose concentration , and urine protein/creatinine ( U-Prot/Cr ) ratio and to compare data with plasma P08253 and -9 activity , cystatin-C and creatinine concentrations in colic horses and healthy controls . Horses with surgical colic ( n = 5 ) were compared to healthy stallions ( n = 7 ) that came for castration . Blood and urine samples were collected . MMP gelatinolytic activity was measured by zymography . RESULTS : We found out that horses with colic had significantly higher urinary P14780 complex and proMMP-9 activities than horses in the control group . Colic horses also had higher plasma P08253 activity than the control horses . Serum creatinine , although within reference range , was significantly higher in the colic horses than in the control group . There was no significant increase in urinary alkaline phosphatase , gamma-glutamyltranspeptidase or total proteins in the colic horses compared to the control group . A human cystatin-C test ( Dako Cytomation latex immunoassay based on turbidimetry ) did not cross react with equine cystatin-C . CONCLUSION : The results indicate that plasma P08253 may play a role in the pathogenesis of equine colic and urinary P14780 in equine kidney damage . Effect of tumor necrosis factor family member O43557 ( O43557 ) on the activation of basophils and eosinophils interacting with bronchial epithelial cells . Allergic asthma can cause airway structural remodeling , involving the accumulation of extracellular matrix and thickening of smooth muscle . P01375 ( P01375 ) family ligand O43557 ( O43557 ) is a cytokine that binds herpesvirus entry mediator ( Q92956 ) / Q92956 and lymphotoxin β receptor ( LTβR ) . O43557 induces asthmatic cytokine P35225 and fibrogenic cytokine transforming growth factor-β release from allergic asthma-related eosinophils expressing Q92956 and alveolar macrophages expressing LTβR , respectively , thereby playing crucial roles in asthmatic airway remodeling . In this study , we investigated the effects of O43557 on the coculture of human basophils/eosinophils and bronchial epithelial BEAS-2B cells . The expression of adhesion molecules , cytokines/chemokines , and matrix metalloproteinases ( MMP ) was measured by flow cytometry , multiplex , assay or ELISA . Results showed that O43557 could significantly promote intercellular adhesion , cell surface expression of intercellular adhesion molecule-1 , release of airway remodeling-related P05231 , P10145 , and P14780 from BEAS-2B cells upon interaction with basophils/eosinophils , probably via the intercellular interaction , cell surface receptors Q92956 and LTβR on BEAS-2B cells , and extracellular signal-regulated kinase , p38 mitogen activated protein kinase , and NF-κB signaling pathways . The above results , therefore , enhance our understanding of the immunopathological roles of O43557 in allergic asthma and shed light on the potential therapeutic targets for airway remodeling .

MH_train_1385

MH_train_1385

MH_train_1385

interacts_with DB00009?

multiple_choice

Pharmacological activities of the MIF-1 analogues Pro- DB00149 - DB00145 , DB00135 -Pro- DB00149 - DB00145 and pareptide . The pharmacological activities of the related free acid analogues of MIF-1 , Pro- DB00149 - DB00145 ( P00747 ) and DB00135 -Pro- DB00149 - DB00145 ( YPLG ) , were investigated because of the possibility that they may be formed during the digestion of milk and wheat proteins in vivo . The amino acid sequences - DB00135 -Pro- DB00149 - DB00145 - and -Pro- DB00149 - DB00145 - are present in proteins from these foods . Chronic administration of either P00747 ( 0.25 mg/kg , SC , P55957 ) or the control substance , pareptide ( 0.25 mg/kg , SC , P55957 ) , antagonized the development of tolerance to the cataleptic effects of haloperidol in mice . The effect of YPLG ( 0.25 mg/kg , SC , P55957 ) on the development of this tolerance was borderline and not statistically significant . Nanomolar concentrations of P00747 , YPLG , and pareptide each increased the in vitro binding of 3H-apomorphine to rat striatal receptors . In this in vitro system , bell-shaped dose response curves were observed for each peptide . The effects of these peptides on tolerance development and apomorphine binding are similar to those previously reported for MIF-1 and demonstrate that amidation at the carboxyl terminus is not required for biological activity . Plasmodium falciparum dihydrofolate reductase alleles and pyrimethamine use in pregnant Ghanaian women . Drug resistance in Plasmodium falciparum affects prevention of malaria in pregnancy . In a cross-sectional study of 530 pregnant Ghanaian women , P. falciparum dihydrofolate reductase ( P00374 ) gene mutations linked with pyrimethamine resistance were assessed and associations with pyrimethamine intake were analyzed . P. falciparum infected 69 % of women without pyrimethamine use , 59 % of those who had a history of pyrimethamine consumption but a negative urine test , and 53 % of individuals with a positive urine test . Eighty-one percent , 43 % , and 74 % of the isolates contained the mutations DB00174 -108 , DB00167 -51 , and DB00125 -59 , respectively . DB00156 -108 occurred in 8 % . DB00205 use was associated with increased frequencies of DB00174 -108 and DB00125 -59 but not of DB00167 -51 or DB00156 -108 . In women with prophylaxis , wild-type parasites were absent and anemia tended to be more common with an increasing number of P00374 gene mutations . DB00205 appears to be not adequately effective in this part of Ghana , most likely due to the predominance of resistant parasites . Selection for resistance following insufficient prophylaxis could possibly affect the efficacy of future intermittent sulfadoxine-pyrimethamine treatment . Activation of spinal alpha-7 nicotinic acetylcholine receptor attenuates remifentanil-induced postoperative hyperalgesia . The activation of alpha-7 nicotinic acetylcholine receptors ( α7-nAchRs ) are currently being considered as novel therapeutic approaches for managing hyperalgesia in inflammation and chronic neuropathic pain , but the role of a7-nAChRs on opioids induced hyperalgesia remain unknown . The present study investigated the effects of α7-nAChRs selective agonists PHA-543613 and type II positive allosteric modulators ( PAMs ) PNU-120596 in remifentanil induced postoperative hyperalgesia . As the results shown , intrathecal treatment with both α7-nAChRs agonists and type II PAMs could attenuate remifentanil induced hyperalgesia by increasing paw withdrawal mechanical threshold ( PWMT ) and paw withdrawal thermal latency ( PWTL ) . Furthermore , we also investigated the protein level of proinflammatory cytokines and phosphorylation N-methyl-d-aspartate receptor 2B subunit ( p- Q13224 ) in the spinal cord . Our data indicated that activation of α7-nAchRs decreased the proinflammatory cytokines ( P01375 -α , P05231 ) and p- Q13224 protein level in the spinal cord . The depression of the increased levels of proinflammatory cytokines and p- Q13224 after remifentanil treatment may contribute to the anti-hyperalgesia effects of PHA-543613and PNU-120596 via α7-nAChRs . Therefore , our findings demonstrated that α7-nAChRs may be potential candidates for treating opioids induced hyperalgesia . The Effect of Xuefuzhuyu Oral Liquid on DB00945 Resistance and Its Association with rs5911 , rs5787 , and rs3842788 Gene Polymorphisms . DB00945 should be continued indefinitely in patients after interventional therapy , but 10 % to 40 % of patients experience recurrent vascular events despite adequate aspirin therapy , a condition known as aspirin resistance ( AR ) . Xuefuzhuyu oral liquid , derived from the classic recipe Xuefuzhuyu decoction , has been well documented to inhibit platelet aggregation and to improve hemorheology . The aims of this study were to investigate the effects of Xuefuzhuyu oral liquid on AR in patients with chronic stable angina after percutaneous coronary intervention ( P05154 ) and the possible genetic markers related to the drug response . 43 patients diagnosed as having aspirin resistance or semi-resistance were randomly divided into control and treatment groups after screening 207 stable Q8NE62 patients . Platelet aggregation rate was determined using turbidimetry . Three single nucleotide polymorphisms in P23219 ( rs5787 , rs3842788 ) and GP IIb ( rs5911 ) were genotyped in whole blood samples using ABI Q96FA3 7900 HT Fast Real-Time instrument and ABI Q96FA3 3730 DNA Sequencer . The results showed that Xuefuzhuyu oral liquid could effectively improve blood stasis syndrome and AR by inhibiting ADP-induced platelet aggregation and that patients with the rs5911 genetic variant exhibited better drug response upon treatment with Xuefuzhuyu oral liquid , which suggests Xuefuzhuyu oral liquid as a new possible drug for the prevention of AR . DB08810 protects against ethanol-induced gastric mucosal injury in rats : role of 5-hydroxytryptamine , prostaglandins and sulfhydryl compounds . This study was designed to determine the gastroprotective properties of cinitapride ( CNT ) , a novel prokinetic benzamide derivative agonist of Q13639 and 5-HT1 receptors and 5-HT2 antagonist , on mucosal injury produced by 50 % ( v/v ) ethanol . Results were compared with those for 5-hydroxytryptamine ( 5-HT : 10 mg kg-1 ) . The possible involvements of gastric mucus secretion , endogenous prostaglandins ( PGs ) and sulfhydryl compounds ( SH ) in the protection mediated by CNT were also examined . Intraperitoneal administration of CNT ( 0.50 and 1 mg kg-1 ) , 30 min before ethanol , significantly prevented gastric ulceration and increased the hexosamine content of gastric mucus . CNT ( 1 mg kg-1 ) also produced a significant increase in gastric mucosal levels of DB00917 , but did not induce any significant changes in SH values . On the contrary , pretreatment with 5-HT worsened ethanol-induced erosions , however , did not affect gastric mucus secretion , glycoprotein content or DB00917 levels , although the non-protein SH fraction was significantly decreased . The present results demonstrate that the gastroprotective effects of CNT could be partly explained by a complex PG dependent mechanism . We suggest that 5-HT dependent mechanisms through 5-HT2 receptor blockade and 5-HT1 receptor activation could be also involved . P62158 -mediated effects of loperamide on chloride transport by brush border membrane vesicles from human ileum . We investigated whether the synthetic opiate loperamide-HCl is able to regulate specific transport systems for sodium and chloride in brush border membrane vesicles ( BBMVs ) from human ileum and whether such activities are mediated by calcium/calmodulin . In BBMVs we studied Na+/H+ antiport , Cl+/OH- antiport , Na+/Cl- cotransport , and the Cl- conductive pathway . Brush border membrane vesicles were incubated with 10 microM loperamide over 4 h at 5 degrees C before the uptake experiments . In ileal BBMVs , loperamide stimulated intravesicular accumulation of Na+ in the presence of Cl- and vice versa . After 1 min of incubation , the stimulatory effect was 35 % +/- 5 % ( p less than 0.005 ) of the control without loperamide . DB00836 also stimulated Cl-/OH- antiport by 30 % +/- 5 % ( p less than 0.005 ) in BBMVs of ileum . In addition , we studied the role of Ca2+/calmodulin in the action of loperamide on chloride transport by human BBMVs . In loperamide-pretreated BBMVs , calmodulin activity was significantly decreased ( 12 +/- 2 vs. 38 +/- 4 pmol/mg protein ) . When loperamide-pretreated vesicles were incubated with 2 microM calcium ( free concentration ) plus 5 microM calmodulin for 1 h at 5 degrees C , complete inhibition of the stimulatory effect of loperamide on Cl-/OH- antiport and Na+/Cl- cotransport was observed . Increasing the Ca2+/calmodulin activity of loperamide-pretreated BBMVs with 2 microM calcium plus 5 microM calmodulin led to a significant inhibition of Cl-/OH- antiport and Na+/Cl- cotransport by 40 % +/- 5 % ( p less than 0.005 ) . Brominated cyclodipeptides from the marine sponge Geodia barretti as selective 5-HT ligands . The brominated cyclodipeptides barettin ( cyclo[(6-bromo-8-entryptophan)arginine] ) and 8,9-dihydrobarettin ( cyclo[(6-bromotryptophan)arginine] ) isolated from the marine sponge Geodia barretti have previously been shown to inhibit settlement of barnacle larvae in a dose-dependent manner in concentrations ranging from 0.5 to 25 microM . To further establish the molecular target and mode of action of these compounds , we investigated their affinity to human serotonin receptors . The tryptophan residue in the barettins resembles that of endogenous serotonin [ 5-hydroxytryptamine ] . A selection of human serotonin receptors , including representatives from all subfamilies ( 1-7 ) , were transfected into P29320 -293 cells . Barettin selectively interacted with the serotonin receptors 5- Q13049 , P28335 , and Q13639 at concentrations close to that of endogenous serotonin , with the corresponding Ki values being 1.93 , 0.34 , and 1.91 microM , respectively . 8,9-Dihydrobarettin interacted exclusively with the P28335 receptor with a Ki value of 4.63 microM ; it failed to show affinity to 5- Q13049 and Q13639 , indicating that the double bond between the tryptophan and arginine residue plays an important role in the interaction with the receptor proteins . Encapsulation of viral vectors for gene therapy applications . In gene therapy , a number of viruses are currently being used as vectors to provide transient expression of therapeutic proteins . A drawback of using free virus is that it gives a potent immune response , which reduces gene transfer and limits re-administration . An alternative delivery system is to encapsulate the virus in poly(lactide-co-glycolide) ( P00747 ) microspheres prior to administration . A recombinant adenovirus ( Ad ) expressing green fluorescent protein ( GFP ) was used to test the transduction efficiency of Ad encapsulated in microspheres on target cells . The number of infected cells that expressed GFP was measured by flow cytometry . It was demonstrated that encapsulated viral vectors could successfully transduce target cells with encapsulation efficiencies up to 23 % and that the level of transduction could be controlled by varying both the quantity of microspheres and the amount of Ad in the microspheres . High transduction efficiencies and its recognized biocompatibility make P00747 -encapsulated Ad an attractive alternative to the use of free virus in gene therapy applications . The infectivity of Ad was found to be significantly influenced by the processing conditions and changes in environmental factors . Free Ad and encapsulated Ad were able to infect both E1 complimenting cells ( P29320 293 ) and non-complimenting cells ( A549 ) , with the viral expression in P29320 293 cells being 2.1 times greater than for A549 cells . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . Upregulation of cell-surface-associated plasminogen activation in cultured keratinocytes by interleukin-1 beta and tumor necrosis factor-alpha . Keratinocytes synthesize and secrete urokinase-type plasminogen activator ( uPA ) which is bound in an autocrine manner to a specific receptor ( uPA-R ) at the keratinocyte surface . P00747 that is also bound to specific membrane binding sites is readily activated by uPA-R-bound uPA . Thus , plasmin is provided for proteolysis of pericellular glycoproteins . The expression of uPA and the uPA-R is confined to migrating keratinocytes during epidermal wound healing , rather than to keratinocytes of the normal epidermis . The regulatory factors of uPA/uPA-R expression in keratinocytes remained largely elusive . Proinflammatory cytokines , such as tumor necrosis factor-alpha ( P01375 ) or interleukin-1 beta ( P01584 ) , are present in epidermal wounds . We have therefore tested P01584 and P01375 for their influence on surface-associated plasminogen activation in a human keratinocyte cell line ( HaCaT ) as well as in primary cultures of normal human epidermal keratinocytes . Both cytokines induced the secretion of uPA into the culture supernatants and a concomitant increase in uPA activity as well as in uPA and uPA-R antigen at the cell surface . The increase was preceded by an increase in specific mRNA . The induction was accompanied by an accelerated uPA-dependent and plasmin-mediated detachment of HaCaT cells from the culture substratum . Taken together , the proinflammatory cytokines P01584 and P01375 induced a coordinated increase in uPA and uPA-R as well as increased pericellular plasmin-mediated proteolysis in human epidermal keratinocytes . This function might be an element of the molecular cell biological events during epidermal wound healing . Cloning and expression of the chromosomal immune interferon gene of the rat . The chromosomal immune interferon gene of the rat ( P01579 ) was identified by screening a recombinant rat lambda phage library with a human P01579 cDNA probe . In contrast to the genes of other rat IFNs , this rat P01579 chromosomal gene contains introns and its structural organization closely resembles that of the human and murine P01579 genes . The rat P01579 gene encodes a signal sequence of 19 amino acids followed by the mature P01579 protein of 137 amino acids . The gene was expressed under control of the simian virus 40 ( SV40 ) early promoter in Chinese hamster ovary ( CHO ) cells deficient in dihydrofolate reductase ( P00374 ) after co-transformation with a plasmid containing the mouse P00374 gene . Initial transformants with a P00374 + phenotype produced P01579 titres ranging from 20 to 1600 units/ml . After stepwise increases in the concentration of methotrexate ( MTX ) in the growth medium of transformed CHO cells , MTX-resistant clones producing 80 000-100 000 units per ml were isolated . Protein analysis of supernatants of these MTX-resistant cells by polyacrylamide gel electrophoresis revealed a product with an apparent mol . wt. of 18 000 daltons which was not detectable in the growth medium of P00374 + transformants that did not produce IFN . The product was identified as rat P01579 and constituted approximately 5 % of the proteins excreted from these cells . Death receptors 4 and 5 activate Nox1 NADPH oxidase through riboflavin kinase to induce reactive oxygen species-mediated apoptotic cell death . Stimulation of the proapoptotic tumor necrosis factor ( P01375 ) -related apoptosis-inducing ligand ( P50591 ) receptors , death receptors 4 ( DR4 ) and 5 ( DR5 ) , conventionally induces caspase-dependent apoptosis in tumor cells . Here we report that stimulation of DR4 and/or DR5 by the agonistic protein KD548-Fc , an Fc-fused DR4/DR5 dual-specific Kringle domain variant , activates plasma membrane-associated Nox1 NADPH oxidase to generate superoxide anion and subsequently accumulates intracellular reactive oxygen species ( ROS ) , leading to sustained c-Jun N-terminal kinase activation and eventual apoptotic cell death in human HeLa and Jurkat tumor cells . KD548-Fc treatment induces the formation of a DR4/DR5 signaling complex containing riboflavin kinase ( Q969G6 ) , Nox1 , the Nox1 subunits ( Rac1 , Noxo1 , and Noxa1 ) , P01375 receptor-associated death domain ( Q15628 ) , and Q12933 ( TRAF2 ) . Depletion of Q969G6 , but not the Nox1 subunits , Q15628 and TRAF2 , failed to recruit Nox1 and Rac1 to DR4 and DR5 , demonstrating that Q969G6 plays an essential role in linking DR4/DR5 with Nox1 . Knockdown studies also reveal that Q969G6 , Q15628 , and TRAF2 play critical , intermediate , and negligible roles , respectively , in the KD548-Fc-mediated ROS accumulation and downstream signaling . Binding assays using recombinantly expressed proteins suggest that DR4/DR5 directly interact with cytosolic Q969G6 through Q969G6 -binding regions within the intracellular death domains , and Q15628 stabilizes the DR4/DR5- Q969G6 complex . Our results suggest that DR4 and DR5 have a capability to activate Nox1 by recruiting Q969G6 , resulting in ROS-mediated apoptotic cell death in tumor cells . Effects of ifenprodil on the discriminative stimulus effects of cocaine in rhesus monkeys . DB08954 is a non-competitive N-methyl-D-aspartate ( DB01221 ) receptor antagonist which prefers Q13224 -containing DB01221 receptors to Q12879 -containing DB01221 receptors . It has been reported that ifenprodil suppresses morphine-induced place preference in mice . In this study , the effects of ifenprodil on the discriminative stimulus effects of cocaine were examined in rhesus monkeys . Five monkeys were trained to discriminate cocaine at 0.25 or 0.5 mg/kg im from saline using a standard two-lever drug-discrimination paradigm under a fixed-ratio schedule of food reinforcement . A single dose of cocaine ( 0.06-0.5 mg/kg ) produced a dose-dependent increase in cocaine-appropriate response , and training doses produced 100 % cocaine-lever response in each monkey . Pretreatment with ifenprodil ( 1 or 2 mg/kg , i.v. ) blocked the cocaine-appropriate response when low doses of cocaine were used . The results suggest that Q13224 -containing DB01221 receptor-mediated mechanisms modulate the discriminative stimulus effects of cocaine in rhesus monkeys . Clinical development of eniluracil : current status . DB03516 is a potent inactivator of dihydropyrimidine dehydrogenase ( Q12882 ) , which is the first enzyme in the degradative pathway of systemically administered 5-fluorouracil ( DB00544 ) . Two completely oral regimens of eniluracil plus DB00544 are being evaluated in clinical trials : ( 1 ) a chronic schedule with both agents administered P55957 in a 10:1 ratio for 28 days of a 5-week course , and ( 2 ) a 5-day schedule of eniluracil once daily on days 1 through 7 and DB00544 once daily on days 2 through 6 . The clinical development of eniluracil is being pursued in several tumor types , including colorectal cancer , breast cancer , and pancreatic cancer . Response rates achieved in a phase II study of the chronic schedule of oral eniluracil/ DB00544 in patients with colorectal cancer compare favorably with those obtained in trials of intravenous DB00544 and leucovorin , while results from other trials are awaited . Safety analysis for the 28-day schedule has revealed a low incidence of severe toxicities , particularly as compared with standard DB00544 regimens . Targeting nanomedicines in the treatment of Crohn 's disease : focus on certolizumab pegol ( DB08904 ) . A variety of targets for therapeutic intervention are based upon advances in understanding of the immunopathogenesis of Crohn 's disease . Crohn 's disease is initiated by an innate immune response , which eventuates in a T-cell driven process , characterized by a T-helper cell 1 type cytokine profile . Several new treatments now focus on suppressing T-cell differentiation or T-cell inflammation . Since inflammatory bowel disease ( Q9UKU7 ) represents a state of dysregulated inflammation , drugs that augment the anti-inflammatory response have the potential to downregulate inflammation and thereby hopefully modify the disease . Tumour necrosis factor ( P01375 ) is a major target of research and clinical investigation . P01375 has proinflammatory effects in the intestinal mucosa and is a pivotal cytokine in the inflammatory cascade . DB08904 ( DB08904 ) is a PEGylated , Fab ' fragment of a humanized anti- P01375 monoclonal antibody . PEGylation increases the half-life , reduces the requirement for frequent dosing , and possibly reduces antigenicity as well . Certolizumab has been shown in Phase III trials to achieve and maintain clinical response and remission in Crohn 's disease patients . It improves the quality of life . DB08904 will be indicated for moderately to severely active Crohn 's disease , but it is not yet licensed in Europe or the US . It is not possible to construct an algorithm for treatment , but when compared with infliximab the two principal advantages are likely to be lower immunogenicity ( as shown by anti-drug antibodies , absence of infusion reactions , and low rate of antinuclear antibodies ) , and a subcutaneous route of administration . These two factors may be sufficient to promote it up the pecking order of anti- P01375 agents . Synthetic delivery system for tuberculosis vaccines : immunological evaluation of the M. tuberculosis 38 kDa protein entrapped in biodegradable P00747 microparticles . Tuberculosis remains a major public health burden which could be ameliorated by effective and well-defined subunit vaccines , particularly because the protective efficacy of current M. bovis BCG vaccines is both unpredictable and variable . The immunodominant 38 kDa antigen from Mycobacterium tuberculosis was entrapped in biodegradable poly ( DL-lactide co-glycolide ) ( P00747 ) microparticles which served as a delivery system . Both cellular and humoral immune responses were assessed and compared with those obtained after immunization with the 38 kDa protein emulsified in incomplete Freund 's adjuvant ( IFA ) . Vaccination of mice with a single dose of antigen-loaded microparticles resulted in specific IgG titres peaking after five weeks comparable to those achieved after vaccination with protein emulsified in incomplete Freund 's adjuvant ( IFA ) . T-cell responses were found to be superior to those induced with antigen/IFA . The T- and B-cell epitope specificities ad judged with synthetic peptides were identical following immunization with antigen in microparticles or IFA . Differences in adjuvanticity were revealed by measuring antigen-specific IgG1 , IgG2a and antigen-induced P01579 secretion in vitro : substantially higher titres of IgG2a were observed following immunization with antigen/microparticles than with 38 kDa protein/IFA . This was paralleled by a tenfold higher secretion of P01579 in mice injected with antigen/microparticles . Reduction in colony-forming units was not consistent in mice immunized with 38 kDa protein entrapped in microparticles which were subsequently infected with live tubercle bacilli . Taken together these results indicate that biodegradable P00747 microparticles constitute a favorable candidate vaccine delivery system worthy of further assessment in the quest to develop better and defined agents protecting against tuberculosis . P15121 regulates high glucose-induced ectodomain shedding of tumor necrosis factor ( P01375 ) -alpha via protein kinase C-delta and P01375 converting enzyme in vascular smooth muscle cells . Chronic low-grade inflammation has emerged as a key contributor to the cardiovascular complications of diabetes , however , the mechanisms by which diabetes increases inflammation remain poorly understood . Here , we report that exposure to high glucose ( HG ) stimulates ectodomain shedding of P01375 from rat aortic smooth muscle cells in culture . Our results show that exposure to HG decreases membrane-associated P01375 . This decrease in unprocessed P01375 was prevented by the aldose reductase ( AR ) inhibitor sorbinil and AR small interference RNA . Treatment with HG , but not equimolar mannitol or 3-O-methyl glucose , resulted in phosphorylation and activation of P01375 converting enzyme ( P78536 ) ( P78536 ) , which were attenuated by sorbinil or AR-specific small interference RNA . HG-induced P78536 phosphorylation and P01375 processing were also prevented by P01375 protease inhibitor-1 , an inhibitor of P78536 . Inhibition of protein kinase C ( PKC ) -delta by rottlerin prevented HG-induced P78536 activation and the accumulation of unprocessed P01375 . Treatment with sorbinil decreased elevated levels of circulating P01375 in streptozotocin-treated diabetic rats . DB02712 treatment also decreased the expression of P01375 , matrix metalloproteinase-2 , matrix metalloproteinase-9 , and increased tissue inhibitor of metalloproteinase-3 in vascular smooth muscle cells treated with HG and in balloon-injured carotid arteries of diabetic rats . These results indicate that HG-induced P01375 shedding could be attributed to P78536 activation , which is regulated , in part , by PKC-delta and AR . Therefore , inhibition of P78536 by P01375 protease inhibitor-1 , or pharmacological inhibition of PKC-delta or AR may represent useful strategies for treating vascular inflammation associated with diabetes . Preparation and characterization of RGD tumour-homing-peptide-modified plasminogen P13647 . P00747 P13647 ( kringle 5 ) has strong inhibitory effects on endothelial-cell proliferation and migration . It was reported that P13647 can reduce tumour neovascularization , resulting in clinically relevant antitumour effects . To determine whether addition of a tumour-targeting peptide could improve the tumour homing and antitumour activities of P13647 , we genetically modified P13647 with an RGD ( DB00125 - DB00145 - DB00128 ) motif , which is a ligand with high affinity for αvβ₃ and αvβ₅ integrins . The fusion protein RGD- P13647 was expressed in the Pichia pastoris system and the biological activity of RGD- P13647 was assessed in vitro and in vivo . The results showed that the RGD- P13647 exhibited a more potent effect of inhibiting endothelial cell proliferation and migration compared with that of traditional P13647 . RGD- P13647 also displayed stronger anti-angiogenic activity in a P62158 ( chick chorioallantoic membrane ) assay . Furthermore , RGD- P13647 also showed stronger anti-angiogenic and antitumour effects in B16F10 melanoma-bearing mice compared with traditional P13647 . In conclusion , the biological activity of P13647 can be further improved by the addition of a tumour-homing peptide , and the RGD- P13647 may prove to be a promising novel candidate for cancer therapy . Interferon-gamma and interleukin 4 inhibit interleukin 1beta-induced delayed prostaglandin E(2)generation through suppression of cyclooxygenase-2 expression in human fibroblasts . Interleukin (IL-)1 stimulates prostaglandin E(2)(PGE(2)) generation in fibroblasts , and preferential couplings between particular phospholipase A(2)(PLA(2)) and cyclooxygenase ( P36551 ) isozymes are implicated with IL-1-induced delayed PGE(2)generation . The regulatory effects of interferon ( IFN ) -gamma and P05112 on IL-1beta-induced P36551 , PLA(2)isoforms expression and terminal delayed PGE(2)generation were examined in three types of human fibroblasts . These human fibroblasts constitutively expressed cytosolic PLA(2)(cPLA(2)) and P23219 enzymes , and exhibited delayed PGE(2)generation in response to IL-1beta . IL-1beta also stimulated expression of cPLA(2)and P35354 only , while constitutive and IL-1beta-induced type IIA and type V secretory PLA(2)s ( sPLA(2)s ) expression could not be detected . A P35354 inhibitor and cPLA(2)inhibitor markedly suppressed the IL-1beta-induced delayed PGE(2)generation , while a type IIA sPLA(2)inhibitor failed to affect it . P01579 and P05112 dramatically inhibited the IL-1beta-induced delayed PGE(2)generation ; these cytokines apparently suppressed IL-1beta-stimulated P35354 expression and only weakly suppressed cPLA(2)expression in response to IL-1beta . These results indicate that IL-1beta-induced delayed PGE(2)generation in these human fibroblasts mainly depends on de novo induction of P35354 and cPLA(2) , irrespective of the constitutive presence of P23219 , and that P01579 and P05112 inhibit IL-1beta-induced delayed PGE(2)generation by suppressing , predominantly , P35354 expression . Role of key residues at the flavin mononucleotide ( Q68DA7 ):adenylyltransferase catalytic site of the bifunctional riboflavin kinase/flavin adenine dinucleotide ( DB03147 ) Synthetase from Corynebacterium ammoniagenes . In mammals and in yeast the conversion of DB00140 ( RF ) into flavin mononucleotide ( Q68DA7 ) and flavin adenine dinucleotide ( DB03147 ) is catalysed by the sequential action of two enzymes : an DB00171 :riboflavin kinase ( Q969G6 ) and an DB00171 : Q8NFF5 ( FMNAT ) . However , most prokaryotes depend on a single bifunctional enzyme , DB03147 synthetase ( FADS ) , which folds into two modules : the C-terminal associated with Q969G6 activity and the N-terminal associated with FMNAT activity . Sequence and structural analysis suggest that the 28-HxGH-31 , 123-Gx(D/N)-125 and 161-xxSSTxxR-168 motifs from FADS must be involved in DB00171 stabilisation for the adenylylation of Q68DA7 , as well as in DB03147 stabilisation for DB03147 phyrophosphorolysis . Mutants were produced at these motifs in the Corynebacterium ammoniagenes FADS ( CaFADS ) . Their effects on the kinetic parameters of CaFADS activities ( Q969G6 , FMNAT and DB03147 pyrophosphorilase ) , and on substrates and product binding properties indicate that H28 , H31 , N125 and S164 contribute to the geometry of the catalytically competent complexes at the FMNAT-module of CaFADS . Successful thrombolysis of a stroke with a pulmonary embolism in a young woman . BACKGROUND : Paradoxical embolism is a rare event , accounting for < 2 % of all arterial emboli . The diagnosis is often difficult , and consequences for the patient can be severe . CASE REPORT : We describe the case of a 35-year-old female physician who presented to our Emergency Department ( ED ) in severe hemodynamic compromise , with an altered level of consciousness and major expressive aphasia 1 day after undergoing a leg varicosal stripping procedure under regional anesthesia . She was successfully thrombolyzed with 0.9 mg/kg of Recombinant Tissue P00747 Activator ( rtPA , DB00009 ) and had a full recovery . CONCLUSION : To our knowledge , this is the first description of a case of massive pulmonary embolism associated with a paradoxical stroke related to patent foramen ovale that was thrombolyzed for both conditions with a " neurological dose " of rtPA . Although thrombolysis was completely successful in this case , indications and contraindications should be thoroughly respected . A more conservative approach with anticoagulation , or a more aggressive approach with surgical thrombectomy , can each potentially have a place in particular cases . Intra-arterial catheter-directed thrombolysis and percutaneous embolectomy are additional options to be considered when available , especially if there are contraindications for systemic thrombolysis .

MH_train_1386

MH_train_1386

MH_train_1386

interacts_with DB06271?

multiple_choice

[ Activation of coagulation cascade in children during an idiopathic nephrotic syndrome relapse ] . The objective of this study was to assess concentrations of selected markers of coagulation in children with relapse of idiopathic nephrotic syndrome during a 6-week therapy . Study groups : 22 subjects ( 32 relapses ) -- 14 males , 8 females ( mean age 7.15 +/- 1.5 y. ) with no thrombotic complications were included into the study . All children were clinically steroid-sensitive . METHODS : Coagulation markers ( platelet count , thrombin time , APTT , INR , fibrinogen 1 + 2 fragments ( F1 + 2 ) , thrombin-antithrombin complexes ( TAT ) , serum levels of D-dimer ( DD ) , fibrin monomers ( FM ) and antithrombin activity ( P01008 ) ) were measured three times : on admission , after 2 and 6 weeks . The control group consisted of 13 healthy children . RESULTS : Serum concentration of TAT or F1 + 2 did not differ between 3 stages ( p > 0.05 ) . However , values at 0 and 2 weeks were significantly higher than in control group ( p < 0.05 ) . We found no correlation between TAT or F1 + 2 and FBG , ALB , TCH , TG levels . [ table : see text ] CONCLUSIONS : The coagulation cascade in relapse of NS was activated during first 6 weeks of therapy whereas metabolic disturbances ( low ALB , high P02675 , TCH , TG , high platelets ) normalized . It is speculative whether it was caused by active immunological process but definitely it resulted in " prothrombotic state " in P01308 patients . Recombinational and physical mapping of the locus for primary open-angle glaucoma ( Q99972 ) on chromosome 1q23-q25 . Primary open-angle glaucoma ( POAG ) is a leading cause of irreversible blindness in industrialized countries . A locus for juvenile-onset POAG , Q99972 , has been mapped to 1q21-q31 in a 9-cM interval . With recombinant haplotypes , we have now reduced the Q99972 interval to a maximum of 3 cM , between the D1S452/NGA1/D1S210 and NGA5 loci . These loci are 2.8 Mb apart on a 4.7-Mb contig that we have completed between the D1S2851 and D1S218 loci and that includes 96 YAC clones and 48 STSs . The new Q99972 interval itself is now covered by 25 YACs , 30 STSs , and 16 restriction enzyme site landmarks . The lack of a NotI site suggests that the region has few CpG islands and a low gene content . This is compatible with its predominant cytogenetic location on the 1q24 G-band . Finally , we have excluded important candidate genes , including genes coding for three ATPases ( P05026 , P23634 , P50993 ) , an ion channel ( VDAC4 ) , antithrombine III ( P01008 ) , and prostaglandin synthase ( P35354 ) . Our results provide a basis to identify the Q99972 gene . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP . P50579 is required for P19526 initiation and proliferation . In a chemical screening , we tested the antiangiogenic effects of fumagillin derivatives and identified fumagillin as an inhibitor of definitive hematopoiesis in zebrafish embryos . DB02640 is known to target methionine aminopeptidase II ( MetAP2 ) , an enzyme whose function in hematopoiesis is unknown . We investigated the role of MetAP2 in hematopoiesis by using zebrafish embryo and human umbilical cord blood models . Zebrafish metap2 was expressed ubiquitously during early embryogenesis and later in the somitic region , the caudal hematopoietic tissue , and pronephric duct . metap2 was inhibited by morpholino and fumagillin treatment , resulting in increased mpo expression at 18 hours postfertilization and reduced c-myb expression along the ventral wall of dorsal aorta at 36 hours postfertilization . It also disrupted intersegmental vessels in Tg(fli1:gfp) embryos without affecting development of major axial vasculatures . Inhibition of MetAP2 in CB P28906 (+) cells by fumagillin had no effect on overall clonogenic activity but significantly reduced their engraftment into immunodeficient nonobese diabetes/severe combined immunodeficiency mice . metap2 knock-down in zebrafish and inhibition by fumagillin in zebrafish and human CB P28906 (+) cells inhibited P62158 Kinase II activity and induced P29323 phosphorylation . This study demonstrated a hitherto-undescribed role of MetAP2 in definitive hematopoiesis and a possible link to noncanonical Wnt and P29323 signaling . DB08816 as an alternative in clopidogrel-associated neutropenia . DB00945 in combination with platelet Q9H244 receptor blocker has become the mainstay antiplatelet treatment strategy for the prevention of stent thrombosis . DB00208 was the first widely used Q9H244 receptor blockers , but clopidogrel has mostly replaced the use of ticlopidine due to its more favorable adverse event profile on bone marrow . However , when clopidogrel induced bone marrow toxicity occurs , little is known about the efficacy and safety of alternative treatments , and thus , in these cases , medical decisions may be very difficult . We report a case of clopidogrel-induced severe neutropenia in a patient treated with coronary stent and safety of alternative treatment with ticagrelor . Modulation of acute graft-versus-host-disease after allogeneic bone marrow transplantation by tumor necrosis factor alpha ( P01375 alpha ) release in the course of pretransplant conditioning : role of conditioning regimens and prophylactic application of a monoclonal antibody neutralizing human P01375 alpha ( DB04956 ) . Contribution of host-related cytokine release in the course of pretransplant conditioning to early tissue damage and induction of acute graft-versus-host disease ( GVHD ) after allogeneic bone marrow transplantation ( BMT ) has been shown in experimental models . We performed a clinical phase I/II trial applying a monoclonal antibody neutralizing human tumor necrosis alpha ( P01375 alpha ) during pretransplant conditioning as additional prophylaxis in high-risk patients admitted to allogeneic BMT ; P01375 alpha serum levels and clinical courses in 21 patients receiving anti- P01375 alpha prophylaxis were compared with data from 22 historical controls . Absence of significant release of P01375 alpha in the period of busulphan ( BUS ) treatment , but significant induction of P01375 alpha by total body irradiation ( TBI ) and cyclophosphamide ( CY ) conditioning were correlated with significantly earlier onset of acute GVHD in patients receiving TBI/CY regimens as compared with BUS/CY-treated patients . Prophylactic application of monoclonal anti- P01375 alpha seemed to postpone onset of acute GVHD from day 15 to day 25 ( P < .05 ) after TBI/CY and from day 33 to day 53 after BUS/CY ( P < .10 ) conditioning . Application of monoclonal anti- P01375 alpha in low and intermediate doses was safe and not associated with an increased incidence of infectious or hematologic complications . Thus , our data provide indirect and direct evidence for involvement of conditioning-related cytokine release in induction of early acute GVHD in the clinical setting and support further investigation of this novel approach in randomized trials . Modulation of mitogen-activated protein kinase cascades by differentiation-1 protein : acquired drug resistance of hormone independent prostate cancer cells . PURPOSE : The inhibitor of differentiation-1 protein ( Id-1 ) is over expressed in multidrug resistance prostate cancer cells . We determined the effect of Id-1 expression and its underlying pathways on the development of multidrug resistance in prostate cancer . MATERIALS AND METHODS : P01008 cells were transfected with the Id-1 gene or a blank vector . Id-1 mRNA expression was determined by reverse transcriptase-polymerase chain reaction and Id-1 protein content was detected by immunoblot and flow cytometry . Cellular cytotoxicity was determined by MTT ( microculture 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide ) assay ( Sigma Chemical Co. , St. Louis , Missouri ) . The activation and expression of mitogen-activated protein kinase ( MAPK ) were measured by transactivation assay and Western blotting , respectively . RESULTS : Id-1 overproduction drove P01008 cells to become resistant to chemotherapeutic agents but did not induce mdr-1 gene expression . The p38MAPK and c-jun N-terminal kinase ( JNK ) pathways were suppressed , which correlated with increased Id-1 expression . No significant change in extracellular signal-regulated kinase ( P29323 ) activation was observed in Id-1 transfectants compared with that of P01008 or vector control . Treatment of Id-1 expressing cells with p38MAPK and JNK inhibitors resulted in decreased doxorubicin induced apoptosis . In contrast , Id-1 expressing cells treated with P29323 inhibitor made cells more sensitive to drug induced apoptosis . CONCLUSIONS : Up-regulation of Id-1 was found in prostate cancer multidrug resistant cells . Sustained P29323 activation , and JNK and p38MAPK inhibition by Id-1 in cells may confer drug resistance . These changes in MAPKs could be a mechanism for the acquisition of multidrug resistance in prostate cancer . Genetic polymorphisms of P23945 , P05093 , P04798 , Q9HC96 , P06213 , P05121 genes in adolescent girls with polycystic ovary syndrome . BACKGROUND : Polycystic ovary syndrome ( PCOS ) , whose genetic basis is not completely well understood , is the most common endocrine disorder in women and it typically develops during adolescence . The aim of this study is to investigate the possible association between single nucleotide polymorphisms ( SNPs ) of P23945 , P05093 , P04798 , Q9HC96 , P06213 , P05121 genes and PCOS in adolescent girls . METHODS : DNA samples from forty-four adolescent girls with PCOS and 50 healthy controls were analyzed by PCR-RFLP and direct DNA sequencing to determine the genotypic frequency of 17 different polymorphic loci on the P23945 ( A307T , N680S ) , P05093 ( -34 T/C ) , P04798 ( T6235C ) , Q9HC96 ( 44 , 43 , 19 , 63 ) , P06213 ( exon 17 C/T ) , P05121 ( 4G/5G ) genes . Genotyping of exon 12 ( six polymorphisms ) and intron 12 ( one polymorphism ) of P06213 gene by direct DNA sequencing was performed for the first time in this study . RESULTS : No significant differences were observed in the genotype and allele distributions of above mentioned polymorphisms between cases and control groups . CONCLUSION : Our data does not support an association between SNPs of P23945 , P05093 , P04798 , Q9HC96 , P06213 , P05121 genes and susceptibility to PCOS or related traits in Turkish adolescent girls . Mechanism of inhibition of the P42262 AMPA receptor channel opening by talampanel and its enantiomer : the stereochemistry of the 4-methyl group on the diazepine ring of 2,3-benzodiazepine derivatives . Stereoselectivity of 2,3-benzodiazepine compounds provides a unique way for the design of stereoisomers as more selective and more potent inhibitors as drug candidates for treatment of the neurological diseases involving excessive activity of AMPA receptors . Here we investigate a pair of enantiomers known as DB04982 and its ( + ) counterpart about their mechanism of inhibition and selectivity toward four AMPA receptor subunits or P42261 -4 . We show that DB04982 is the eutomer with the endismic ratio being 14 for the closed-channel and 10 for the open-channel state of P42262 . Kinetic evidence supports that DB04982 is a noncompetitive inhibitor and it binds to the same site for those 2,3-benzodiazepine compounds with the C-4 methyl group on the diazepine ring . This site , which we term as the " M " site , recognizes preferentially those 2,3-benzodiazepine compounds with the C-4 methyl group being in the R configuration , as in the chemical structure of DB04982 . Given that DB04982 inhibits P42261 and P42262 , but is virtually ineffective on the P42263 and P48058 AMPA receptor subunits , we hypothesize that the " M " site(s) on P42261 and P42262 to which DB04982 binds is different from that on P42263 and P48058 . If the molecular properties of the AMPA receptors and DB04982 are used for selecting an inhibitor as a single drug candidate for controlling the activity of all AMPA receptors in vivo , DB04982 is not ideal . Our results further suggest that addition of longer acyl groups to the N-3 position should produce more potent 2,3-benzodiazepine inhibitors for the " M " site . Lack of biological relevance of platelet cyclooxygenase-2 dependent thromboxane A2 production . INTRODUCTION : There is emerging evidence of a considerable variability of the impact of aspirin on clinical outcome and laboratory findings . Persistent TxA2 production seems to be the most likely reason . Aim of this study was to determine whether the mechanism responsible for TxA2 persistent production is , at least partially , dependent upon aspirin-insensitive platelet P35354 enzymatic pathway . METHODS AND RESULTS : In 100 consecutive patients , under chronic aspirin anti-platelet treatment ( 100-160 mg/day ) selected on the basis of detectable plasma salicylate levels , serum and DB04557 ( AA ) -induced platelet TxA2 production , immunoblot analysis of platelet P23219 / P35354 expression and P35354 activity were studied . Immunoblot revealed P35354 expression in 46 % patients , in an amount that was markedly lower than P23219 . In 10 P35354 positive patients with TxA2 levels over the median , AA-induced TxA2 production performed in vitro in the presence of the P35354 inhibitor CAY10404 and aspirin demonstrated that P35354 dependent TxA2 production is less than 2 % . CONCLUSION : Our data demonstrate that the inter-individual variability of platelet sensitivity to aspirin is due to a reduced efficacy of aspirin on platelet P23219 despite ascertained patient compliance . We suggest that serum TxA2 assay might be performed in future clinical studies to improve our knowledge on the residual TxA2 production in aspirin-treated patients . Structure and function of eritadenine and its 3-deaza analogues : potent inhibitors of S-adenosylhomocysteine hydrolase and hypocholesterolemic agents . d- DB03769 ( DEA ) is a potent inhibitor of S-adenosyl-l-homocysteine hydrolase ( P23526 ) and has hypocholesterolemic activity . We have hypothesized that 3-deaza-DEA ( P01024 -DEA ) and its analogues retain high level of P23526 inhibitory activity and have resistance to deamination and glycosidic bond hydrolysis in vivo . Such P01024 -DEA analogues would have much higher hypocholesterolemic activity . P01024 -DEA , and its methyl ester ( P01024 -OMeDEA ) and its methyl amido ( P01024 -NMeDEA ) were synthesized to examine their P23526 inhibitory and hypocholesterolemic activities . A crystal structure of P23526 containing P01024 -DEA was determined and confirmed that DEA and P01024 -DEA bound to the same site of P23526 with the same binding mode . The P23526 inhibitory activities of P01024 -DEA ( K(I)=1.5 microM ) and P01024 -OMeDEA ( K(I)=1.5 microM ) are significantly lower than that of DEA ( K(I)=30 nM ) , while rats fed by P01024 -DEA and P01024 -OMeDEA decrease the total plasma cholesterol and phospholipids by 36-40 % and 23 % , respectively , which is similar to the level of reductions ( 42 % and 27 % ) by DEA . P01024 -NMeDEA lost most of the P23526 inhibitory activity ( K(I)=30 microM ) and dietary P01024 -NMeDEA does not decrease cholesterol and phospholipid in plasma but decreases the triacylglycerol level by 16 % . DEA and P01024 -DEA analogues are neither substrates nor inhibitors of adenosine deaminase . Delineating biological pathways unique to embryonic stem cell-derived insulin-producing cell lines from their noninsulin-producing progenitor cell lines . To identify unique biochemical pathways in embryonic stem cell-derived insulin-producing cells as potential therapeutic targets to prevent or delay beta-cell dysfunction or death in diabetic patients , comparative genome-wide gene expression studies of recently derived mouse insulin-producing cell lines and their progenitor cell lines were performed using microarray technology . Differentially expressed genes were functionally clustered to identify important biochemical pathways in these insulin-producing cell lines . Biochemical or cellular assays were then performed to assess the relevance of these pathways to the biology of these cells . A total of 185 genes were highly expressed in the insulin-producing cell lines , and computational analysis predicted the pentose phosphate pathway ( PPP ) , clathrin-mediated endocytosis , and the peroxisome proliferator-activated receptor ( Q07869 ) signaling pathway as important pathways in these cell lines . P01308 -producing ERoSHK cells were more resistant to hydrogen peroxide ( H(2)O(2) ) -induced oxidative stress . Inhibition of PPP by dehydroepiandrosterone and 6-aminonicotinamide abrogated this H(2)O(2) resistance with a concomitant decrease in PPP activity as measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide ( MTT ) assay . Clathrin-mediated endocytosis , which is essential in maintaining membrane homeostasis in secreting cells , was up-regulated by glucose in ERoSHK but not in their progenitor ERoSH cells . Its inhibition by chlorpromazine at high glucose concentration was toxic to the cells . DB00197 , a P37231 agonist , up-regulated expression of Ins1 and Ins2 but not Glut2 . Gene expression analysis has identified the PPP , clathrin-mediated endocytosis , and the Q07869 signaling pathway as the major delineating pathways in these insulin-producing cell lines , and their biological relevance was confirmed by biochemical and cellular assays . delta- DB00855 dehydratase ( P13716 ) porphyria : the first case in North America with two novel P13716 mutations . The molecular basis of the enzymatic defect responsible for delta-aminolevulinate dehydratase ( P13716 ) porphyria ( ADP ) was investigated in a 14-year-old male who presented clinical and laboratory findings typical of ADP . Nucleotide sequence analysis of P13716 cDNAs from the proband revealed two novel mutations , a 265G to A base transition ( C1 ) and a 394C to T base transition ( P06681 ) , resulting in amino acid substitutions , Glu89Lys and Cys132Arg , respectively . Both mutations were present within exon 5 of the P13716 gene , and appeared to influence the binding of zinc to the enzyme which is essential for enzyme activity . It was found that the C1 mutation was inherited from his father , while the P06681 mutation was from his mother . Expression of these mutant P13716 cDNAs in Chinese hamster ovary cells produced normal P13716 mRNA levels , but markedly decreased P13716 protein and enzyme activity . These results suggest that the combination of the two aberrant ALADs with little enzyme activity accounts for the markedly decreased P13716 activity observed in the proband . This case represents the molecular analysis of the P13716 gene defects in the first case of ADP identified in North America , who is a compound heterozygote for two novel P13716 gene defects . Glycoprotein IIb/IIIa and Q9H244 receptor antagonists yield additive inhibition of platelet aggregation , granule secretion , soluble P29965 release and procoagulant responses . Glycoprotein IIb/IIIa ( P08514 /IIIa ) antagonists , including abciximab and tirofiban , are administered concurrently with clopidogrel , a Q9H244 antagonist , and aspirin in some patients undergoing percutaneous coronary intervention . We studied the effects of , and interactions between , abciximab , tirofiban , aspirin and the Q9H244 antagonist cangrelor on platelet aggregation , alpha and dense granule secretion and procoagulant responses in vitro . Blood was obtained from healthy volunteers . Platelet aggregation , dense granule secretion , alpha granule secretion ( P05121 and soluble P29965 levels ) and procoagulant responses ( annexin-V and microparticle formation ) were assessed using collagen and thrombin receptor activating peptide ( TRAP ) as agonists . All the antagonists used singularly inhibited collagen-induced responses . Combinations of abciximab or tirofiban with aspirin and/or cangrelor gave additive inhibition with the greatest effect seen when abciximab or tirofiban was combined with both aspirin and cangrelor . DB06441 inhibited TRAP-induced responses and , again , there was additive inhibition of these parameters when abciximab or tirofiban were combined with cangrelor . The P08514 /IIIa receptor plays an important role in amplification of platelet activation such that there are important interactions between P08514 /IIIa antagonists and inhibitors of both Q9H244 receptor activation and , to a lesser extent , thromboxane A2 generation . These interactions are likely to have important influences on the safety and efficacy of combination anti-platelet therapies . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . Interaction of plasma proteins with heparinized gel particles studied by high-resolution two-dimensional gel electrophoresis . In order to further the understanding of protein-surface interactions in the coagulation system , we have chosen to study plasma protein adsorption onto heparin-immobilized surfaces . DB01109 -binding proteins are abundant in plasma : a search of amino acid sequences revealed that many plasma proteins have possible heparin binding sites . Plasma protein adsorption to the heparinized surfaces is monitored by a novel technique in which the solution depletion of proteins is analytically determined using quantitative two-dimensional polyacrylamide gel electrophoresis ( 2-D PAGE ) . This method enables simultaneous , quantitative detection of the majority of plasma proteins before , during , and after their adsorption onto high surface area adsorbents . Using computerized densitometry of silver-stained 2-D PAGE gels , the amount of each protein can be determined from the integrated optical density of each protein " spot. " Kinetics of adsorption and adsorption isotherms of four important heparin binding proteins , antithrombin III ( P01008 ) , complement factor P01024 ( P01024 ) , apolipoprotein AI ( P02647 ) and apolipoprotein AIV ( P06727 ) are reported in this paper . From the adsorption isotherms , the apparent binding constants of each protein-immobilized heparin complex , Ka , were calculated . The surface binding constants were of the same order of magnitude as the respective solution binding constants in the literature . The surface binding constants followed the same order as the respective solution binding constants : Ka ( P01008 ) greater than Ka ( P06727 ) greater than Ka ( P01024 ) greater than Ka ( P02647 ) , indicating that protein binding to the immobilized heparin used is not essentially different from solution binding . P09038 selectively increases AMPA-receptor subunit GluR1 protein level and differentially modulates Ca2+ responses to AMPA and DB01221 in hippocampal neurons . The excitatory neurotransmitter glutamate is believed to play important roles in development , synaptic plasticity , and neurodegenerative conditions . Recent studies have shown that neurotrophic factors can modulate neuronal excitability and survival and neurite outgrowth responses to glutamate , but the mechanisms are unknown . The present study tested the hypothesis that neurotrophic factors modulate responses to glutamate by affecting the expression of specific glutamate-receptor proteins . Exposure of cultured embryonic rat hippocampal cells to basic fibroblast growth factor ( P09038 ) resulted in a concentration-dependent increase in levels of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate ( AMPA ) -receptor subunit GluR1 protein as determined by western blot , dot-blot , and immunocytochemical analyses . In contrast , P09038 did not alter levels of GluP2/3 , P48058 , or the DB01221 -receptor subunit Q9UHB4 . Nerve growth factor did not affect GluR1 levels . DB01373 -imaging studies revealed that elevation of [Ca2+]i , resulting from selective AMPA-receptor activation , was enhanced in P09038 -pretreated neurons . On the other hand , [Ca2+]i responses to DB01221 -receptor activation were suppressed in P09038 -treated neurons , consistent with previous studies showing that P09038 can protect neurons against DB01221 toxicity . Moreover , neurons pretreated with P09038 were relatively resistant to the toxicities of glutamate and AMPA , both of which were shown to be mediated by DB01221 receptors . These data suggest that differential regulation of the expression of specific glutamate-receptor subunits may be an important mechanism whereby neurotrophic factors modulate activity-dependent neuronal plasticity and vulnerability to excitotoxicity . Modular synthesis of heparin oligosaccharides . A general , modular strategy for the first completely stereoselective synthesis of defined heparin oligosaccharides is described . Six monosaccharide building blocks ( four differentially protected glucosamines , one glucuronic and one iduronic acid ) were utilized to prepare di- and trisaccharide modules in a fully selective fashion . Installation of the alpha-glucosamine linkage was controlled by placing a conformational constraint on the uronic acid glycosyl acceptors thereby establishing a new concept for stereochemical control . Combination of disaccharide modules to form trans-uronic acid linkages was completely selective by virtue of P06681 participating groups . Coupling reactions between disaccharide modules exhibited sequence dependence . While the union of many glucosamine uronic acid disaccharide modules did not meet any problems , certain sequences proved not accessible . Elaboration of glucosamine uronic acid disaccharide building blocks to trisaccharide modules by addition of either one additional glucosamine or uronic acid allowed for stereoselective access to oligosaccharides as demonstrated on the example of a hexasaccharide resembling the P01008 -binding sequence . Final deprotection and sulfation yielded the fully synthetic heparin oligosaccharides . Antagonism by salvianolic acid B of lipopolysaccharide-induced disseminated intravascular coagulation in rabbits . The aim of the present study was to investigate the effects of salvianolic acid B on lipopolysaccharide ( LPS ) -induced disseminated intravascular coagulation ( DIC ) in rabbits . Continuous infusion of LPS was used to induce a DIC model in rabbits . Treatment with salvianolic acid B ( 1 , 3 or 6 mg/kg ) was started simultaneously with LPS infusion ( 0.5 mg/kg LPS in 60 mL saline ; 10 mL/h over a period of 6 h ) through the contralateral marginal ear vein . Activated partial thromboplastin time ( APTT ) , prothrombin time ( PT ) , platelet count and fibrinogen concentration were determined , as were plasma levels of fibrin-fibrinogen degradation products ( Q9NRC9 ) , alanine aminotransferase ( ALT ) , blood urea nitrogen ( BUN ) , protein C activity , antithrombin III ( P01008 ) and tumour necrosis factor ( P01375 ) -α concentration . The gradual impairment of haemostatic parameters was induced by continuous infusion of LPS . There were marked increases in APTT , PT , BUN , ALT and plasma P01375 -α and marked decreases in the platelet count , fibrinogen , Q9NRC9 , protein C and P01008 . The intravenous administration of 1 , 3 or 6 mg/kg salvianolic acid B attenuated the increases in APTT , PT , BUN , ALT and plasma P01375 -α and the decreases in fibrinogen , platelet , Q9NRC9 , protein C and P01008 induced by LPS infusion . These observations indicate that salvianolic acid B has an effect against LPS-induced DIC in rabbits . Her-2/neu expression is a negative prognosticator in ovarian cancer cases that do not express the follicle stimulating hormone receptor ( P23945 ) . BACKGROUND : Anti-Her-2 treatment is successfully administered to Her-2 overexpressing breast cancer patients and significantly implicates upon their survival . Building on these promising results , anti-Her-2 treatment protocols were tested as an option for epithelial ovarian cancer ( EOC ) as well . However Her-2 signalling is known to be modulated by G-protein coupled receptors ( GPCR ) . Since a common GPCR in ovarian cancer is the DB00094 receptor ( P23945 ) , we investigated the prognostic significance of Her-2 in patients that had been stratified according to their P23945 status . FINDINGS : A total number of 153 EOC patients were included in this study . Her-2 positivity was assessed using a standard protocol . Intriguingly Her-2 turned out to be an independent prognostic marker for poor overall survival only in those patients that did not express P23945 . This did neither apply for the whole panel nor in case of P23945 co-expression . CONCLUSIONS : We thus conclude that Her-2 can be a negative prognosticator only in P23945 negative EOC cases . Hence by stratifying EOC patients according to their P23945 expression status , we introduce a diagnostic protocol to effectively select EOC patients that would most probably respond to anti-Her-2 treatment . This observation could be of clinical importance in terms of selecting the patient that would most likely benefit from anti-Her-2 treatment .

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Internalization of OspA in rsCD14 complex and aggregated forms . Although the spirochetal protein OspA is capable of stimulating immune cells in a P08571 - and O60603 -dependent manner , little is known about how O60603 receptor complex ligands , such as OspA , are handled by the cell once delivered . We examine here the internalization of the fluorescently derivatized forms of both the full length DB00045 delivered as a recombinant soluble P08571 ( rsCD14 ) complex and the corresponding lipohexapeptide given to the cells as an aggregate . Both forms of OspA are internalized in a similar manner to acetylated low density lipoprotein ( AcLDL ) , a scavenger receptor ligand . Acetylated low density lipoprotein is capable of competing for internalization with OspA even when OspA is delivered as a rsCD14 complex . We observe co-localization of OspA with lysosomes but not with the Golgi complex . These phenomena are similar between RAW264.7 macrophages and endothelial cells but change drastically when the cells are deprived of serum . Upon serum starvation , OspA shows some localization to the Golgi apparatus whereas the lipohexapeptide remains on the cell surface . Inhibition of internalization of OspA via treatment with cytochalasin D or of the lipohexapeptide via serum starvation does not interfere with P01375 induction activity , consistent with signalling from the cell surface . Interaction of tacrolimus(FK506) and its metabolites with FKBP and calcineurin . DB00864 (FK506) is a strong immuno-suppressant and shows its activity through inhibiting P60568 mRNA transcription by forming pentameric complex with intracellular receptor ( FK506 binding protein 12 kDa or P62942 ) , Ca2+ , calmodulin , and calcineurin . Here , we report the binding activity to P62942 , the pentameric complex formation and Con-A response inhibiting activities of 7 metabolites . C15-demethylated metabolite(M-3) needed higher quantity to compete in Con-A assay and in pentamer formation assay , although it binds more strongly to P62942 . The result suggests that the ability to form a pentameric complex is not a two step reaction with the first binding to P62942 , but a single step reaction by components for the pentamer formation . Binding affinities for sulfonamide inhibitors with matrix metalloproteinase-2 using a linear response method . Due to their involvement in many pathological conditions , matrix metalloproteinases ( MMPs ) , are very attractive therapeutic targets . Our study focuses on one of them , P08253 , which is involved in tumor progression and metastasis . Recently , the solution structure of the catalytic domain of P08253 complexed with a hydroxamic acid inhibitor ( DB01630 ) was published by Feng et al . Using the Hanessian group published binding affinity data and the structure published by Feng as a basis , we have built a binding affinity model by targeting the S(2) ' pocket of the enzyme with a set of nine alpha-N-sulfonylamino hydroxamic acid derivatives . Two binding geometries of each ligand have been generated corresponding to two binding modes denoted A and B , respectively , of which the first one is targeting the S(2) ' pocket and the second one the S(1) pocket . For the binding affinity model developed for mode A the computed activities show a rmsd of 0.583 kcal/mol as compared with the experimental data , and a correlation coefficient r(2) of 0.779 , while in the case of the binding mode B a rmsd of 0.834 kcal/mol and correlation coefficient r(2) of 0.500 , respectively , were obtained . In conclusion , our data suggest a higher probability for the DB00120 (76) gated S(2) ' open form pocket to accommodate the substituent alpha versus the wide solvent exposed S(1) subsite , probability which some research groups could have overlooked due to extensive use in their calculations of non revealing S(2) ' pocket open state crystallographic structures instead of NMR ones . P43220 agonists for type 2 diabetes mellitus : recent developments and emerging agents . More than 26 million people in the United States have type 2 diabetes mellitus ( T2D ) . Many treatment options exist , but achieving long-term glycemic control in patients with T2D remains challenging . The glucagon-like peptide-1 receptor agonists ( P0C6A0 RAs ) offer a treatment option that improves glycemic control and reduces weight , with a low risk of hypoglycemia . They have emerged as attractive options for the treatment of T2D , and significant advances and developments continue to be published regarding these agents . To identify relevant literature on emerging issues related to P0C6A0 RAs , a search of the MEDLINE database was performed . Studies published in English evaluating the safety and efficacy of P0C6A0 RAs were analyzed . Because of their advantages and unique mechanism of action , P0C6A0 RAs are currently being studied in new clinical areas , including in combination with basal insulin , as adjunctive therapy in type 1 diabetes , and for weight loss . In addition , there are several emerging agents in development . DB09265 is a once-daily P0C6A0 RA that targets postprandial glucose and may be most useful when added to basal insulin as an alternative to rapid-acting insulin . DB09043 and dulaglutide are once-weekly P0C6A0 RAs that may offer more convenient dosing . The most common adverse effects of all P0C6A0 RA agents are gastrointestinal ( e.g. , nausea , diarrhea , and vomiting ) , but the rates of occurrence vary among agents . Due to the differences in pharmacokinetics , efficacy , rates of adverse effects , and administration requirements within the P0C6A0 RA class , each agent should be evaluated independently . The future of P0C6A0 RAs offers broader treatment options for T2D as well as potential in other treatment areas . Overexpression of hepatocyte growth factor receptor in scleroderma dermal fibroblasts is caused by autocrine transforming growth factor β signaling . Cutaneous fibrosis seen in systemic sclerosis ( SSc ) is caused by fibroblast activation and abnormal collagen accumulation due to ' autocrine transforming growth factor ( TGF ) -β/Smad signaling ' . P14210 ( P14210 ) may have therapeutic value against SSc , because of its inducible effect on the expression of matrix metalloproteinase ( MMP ) -1 . Previous studies indicated SSc dermal fibroblasts overexpress P08581 c-met , which suggest specific and effective induction of P03956 in SSc fibroblasts caused by P14210 treatment . However , the exact mechanism of c-met overexpression in SSc cells was hardly investigated . We hypothesized that such c-met overexpression is also caused by autocrine TGF-β/Smad signaling . Expression of c-met protein in cultured SSc dermal fibroblasts was significantly up-regulated compared with that in normal fibroblasts . Ectopic TGF-β stimulation induced c-met synthesis in normal fibroblasts , while a TGF-β knockdown normalized the up-regulated c-met levels in SSc fibroblasts . Furthermore , we found the c-met promoter contains a putative binding site for Smads , and the binding activity of Q15796 /3 to the c-met promoter was constitutively up-regulated in SSc fibroblasts as well as in normal fibroblasts treated with exogenous TGF-β1 . Taken together , c-met may be overexpressed due to autocrine TGF-β/Smad signaling in SSc . Considering that P14210 has an antifibrotic effect , such c-met overexpression in SSc fibroblasts may be a negative feedback against cutaneous fibrosis . Clarifying the mechanisms of c-met overexpression and controlling the P14210 /c-met pathway may lead to a new therapeutic approach for this disease . DB08865 for the treatment of patients with advanced non-small cell lung cancer . DB08865 is a potent small-molecule inhibitor of Q9UM73 ( anaplastic lymphoma kinase ; Q9UM73 ) and hepatocyte growth factor receptor ( P08581 , proto-oncogene c- DB00134 ) . A range of tumors , including subsets of non-small cell lung cancer ( NSCLC ) , anaplastic large cell lymphoma and inflammatory myofibroblastic tumors harbor an Q9UM73 rearrangement that leads to oncogenic activation of Q9UM73 . DB08865 has demonstrated preclinical and clinical activity against such malignancies through inhibition of Q9UM73 , and patients harboring Q9UM73 - rearranged NSCLC have demonstrated high response rates and prolonged progression-free survival in phase I and II studies . In August 2011 , crizotinib was approved for the treatment of advanced Q9UM73 -positive NSCLC . The anti-inflammatory and anti-angiogenic role of DB05914 in corneal wound healing following chemical injury . To investigate the anti-inflammatory and anti-angiogenic effects of DB05914 ( O60682 ) in the chemically burned corneas , we mechanically removed the corneal epithelium of rats after 100 % alcohol instillation . The rats were then randomized into four groups : fresh media , conditioned media derived from the O60682 culture ( O60682 -CM ) , O60682 applied topically to the damaged corneas for 2 hours immediately after the injury or O60682 -CM applied either once or 3 times per day for 3 consecutive days . Corneal surface was evaluated every week . After 3 weeks , the corneas were stained with the hematoxylin-eosin , and the expression of interleukin ( IL ) -2 , interferon ( IFN ) -gamma , P05231 , P22301 , transforming growth factor ( TGF ) -beta1 , thrombospondin-1 ( P07996 -1 ) , matrix metalloproteinase-2 ( P08253 ) , and vascular endothelial growth factor ( P15692 ) were analyzed . P01730 + cells were assessed in the corneas . We found that both O60682 and three-time applied O60682 -CM ( 1 ) reduced corneal inflammation and neovascularization , ( 2 ) decreased P60568 and P01579 , although increased P22301 and TGF-beta1 as well as P05231 , ( 3 ) reduced the infiltration of P01730 + cells , and ( 4 ) upregulated the expression of P07996 -1 , although downregulated that of P08253 . Interestingly , whereas three-time application of O60682 -CM was partially effective , transplantation of O60682 achieved a better outcome in suppressing corneal inflammation . The results of this study suggest that the anti-inflammatory and anti-angiogenic action of O60682 in the chemically burned corneas might be mediated in part through paracrine pathways involving soluble factors such as P22301 , TGF-beta1 , P05231 and P07996 -1 . Heart allograft protection with low-dose carbon monoxide inhalation : effects on inflammatory mediators and alloreactive T-cell responses . BACKGROUND : DB11588 ( CO ) , a byproduct of heme catalysis , has lately received considerable attention as a regulatory molecule in cellular and biological processes . CO has been shown to provide potent protection against a variety of tissue injuries . We hypothesized in this study that low concentration CO would be beneficial for organ allografts , which frequently undergo several types of injury such as ischemia/reperfusion , alloimmune reaction , and inflammation METHODS : The efficacy of low-dose CO was examined in a fully allogeneic LEW to BN rat heterotopic heart transplantation ( HHTx ) model . Recipients were kept in air or exposed to low-dose CO ( 20 ppm ) for 14 , 28 , or 100 days after HHTx under short-course tacrolimus RESULTS : CO treatment ( d0-28 , 0-100 ) was remarkably effective in prolonging heart allograft survival to a median of > 100 from 45 days in the air-control group , with significant reductions of arteritis , fibrosis , and cellular infiltration , including macrophages and T cells . CO inhibited intragraft upregulation of Th1 type cytokines ( P60568 , IFNgamma ) , proinflammatory mediators ( IL-1beta , TNFalpha , P05231 , P35354 ) , and adhesion molecule . Shorter CO exposure in early ( 0-13d ) and late ( 14-28d ) posttransplant periods also prolonged graft survival , with a significant inhibition of inflammatory mediators CONCLUSIONS : These results show that low dose CO inhalation protects heart allografts and can considerably prolong their survival . CO appears to function via multiple mechanisms , including direct inhibition of Th1 type cytokine production and regulation of inflammatory responses . Inhibition of neuronal nitric oxide reduces anxiety-like responses to pair housing . Many psychological disorders are characterized by anxiety and alterations in social interactions . Recent studies demonstrate that the chemical messenger nitric oxide ( NO ) can regulate both anxiety and social behaviours . We tested whether an enzyme that produces NO in the brain , neuronal nitric oxide synthase ( P29475 ) , serves as an interface between social interactions and anxiety-like behaviour . Several investigators have observed that mice increase anxiety-like responses in the elevated plus-maze after pair housing . P29475 gene deletion and DB01997 were used to inhibit the production of neuronal NO . Similar to previous studies , pair housing reduced open arm exploration in the elevated plus-maze . Pair housing also increased corticotropin-releasing hormone ( P06850 ) immunoreactive cells in the paraventricular nucleus ( PVN ) of the hypothalamus . Inhibition of NO production increased open arm exploration in pair-housed mice but decreased open arm exploration in individually housed mice . These results suggest that the effect of P29475 inhibition on anxiety-like responses is context dependent and that behavioural responses to social housing are altered after P29475 inhibition . This research suggests that NO may play an important role in mediating the effect social interactions have on anxiety . Targeting of the P40337 -hypoxia-inducible factor-hypoxia-induced gene pathway for renal cell carcinoma therapy . Treatment of advanced renal cancer has made little progress in the past 30 yr . Most clinical efforts have incorporated cytokine-based therapy . The presumption has been that the cytokines may trigger a host immune response against the renal cancer . Only IFN-alpha and high-dose P60568 seemed to have positive effects on patient outcomes . IFN has prolonged the lives of patients by a few months , and high-dose P60568 is capable of inducing very prolonged remissions ( > 5 yr ) for a small number of patients . Nephrectomy in the presence of metastatic disease has been established as an effective procedure for select patients , providing palliation and prolonging survival . Finally , enthusiasm has focused on the use of nonmyeloablative allogeneic stem cell transplantation and donor leukocyte infusion for the induction of graft versus tumor effects . Early results are both provocative and promising . A number of agents that target the critical gene products downstream from P40337 and hypoxia-inducible factor-1 , such as vascular endothelial growth factor , PDGF , P01133 receptor , and TGF-alpha , have recently become available . The new agents are capable of inhibiting specific cellular targets , and the biologic characteristics of clear cell carcinoma of the kidney support their application . If the correct targets are carefully selected for inhibition in tumors in which the targets are present ( clear cell histologic features and loss of P40337 expression ) , then results should resemble those others have observed with targeted therapy , such as the use of STI-571 ( Gleevec ; Novartis Pharmaceuticals , East Hanover , NJ ) for treatment of chronic myelogenous leukemia and gastrointestinal stromal tumors or anti- P04626 /neu ( Herceptin ; Genentech , South San Francisco , CA ) for treatment of breast cancer . Generation of Epstein-Barr virus-specific cytotoxic T lymphocytes resistant to the immunosuppressive drug tacrolimus ( FK506 ) . Adoptive transfer of autologous Epstein-Barr virus-specific cytotoxic T lymphocytes ( EBV-CTLs ) to solid organ transplant ( SOT ) recipients has been shown safe and effective for the treatment of EBV-associated posttransplantation lymphoproliferative disorders ( PTLDs ) . SOT recipients , however , require the continuous administration of immunosuppressive drugs to prevent graft rejection , and these agents may significantly limit the long-term persistence of transferred EBV-CTLs , precluding their use as prophylaxis . DB00864 ( FK506 ) is one of the most widely used immunosuppressive agents in SOT recipients , and its immunosuppressive effects are largely dependent on its interaction with the 12-kDa FK506-binding protein ( P62942 ) . We have knocked down the expression of P62942 in EBV-CTLs using a specific small interfering RNA ( siRNA ) stably expressed from a retroviral vector and found that P62942 -silenced EBV-CTLs are FK506 resistant . These cells continue to expand in the presence of the drug without measurable impairment of their antigen specificity or cytotoxic activity . We confirmed their FK506 resistance and anti-PTLD activity in vivo using a xenogenic mouse model , suggesting that the proposed strategy may be of value to enhance EBV-specific immune surveillance in patients at high risk of PTLD after transplantation . Local control of alpha1-proteinase inhibitor levels : regulation of alpha1-proteinase inhibitor in the human cornea by growth factors and cytokines . Alpha 1-proteinase inhibitor is a major serine proteinase inhibitor in the human cornea involved in the protection of the avascular corneal tissue against proteolytic damage . This inhibitor is upregulated systemically during infection , inflammation and injury . Cytokines that mediate the acute phase response such as IL-1beta and P60568 increased alpha1-proteinase inhibitor present in corneal organ culture media . This released inhibitor represented mainly newly synthesized protein . However , P05231 , a general inducer of the acute phase response that upregulates alpha1-proteinase inhibitor in all other tissues and cells tested , failed to alter corneal alpha1-proteinase inhibitor levels over the tested period of 24 h . In addition to IL-1beta and P60568 , alpha1-proteinase inhibitor levels in the corneal organ culture medium increased following the addition of P09038 and P05019 . The effect of the above growth factors and cytokines was relatively fast with maximal induction observed within the first 5 h . Among the tested growth factors and cytokines , IL-1beta was the most potent and increased total corneal alpha1-proteinase inhibitor levels approximately 2.4-fold in the cornea organ culture medium . Newly , synthesized alpha1-proteinase secreted into the medium increased 3.9-fold . In addition to the effect on corneal alpha1-proteinase inhibitor , IL-1beta also increased the amount of alpha1-proteinase inhibitor released by monocytes and macrophages but not by HepG2 , CaCo2 , and MCF-7 cells within 24 h . These results suggest that the cornea can locally control levels of alpha1-proteinase inhibitor in response to an inflammatory insult . Signaling by proinflammatory cytokines : oligomerization of TRAF2 and Q9Y4K3 is sufficient for JNK and IKK activation and target gene induction via an amino-terminal effector domain . Interleukin-1 ( IL-1 ) and tumor necrosis factor ( P01375 ) stimulate transcription factors AP-1 and NF-kappaB through activation of the Q96HU1 kinases JNK and p38 and the O15111 ( IKK ) , respectively . The P01375 and IL-1 signals are transduced through TRAF2 and Q9Y4K3 , respectively . Overexpressed TRAF2 or Q9Y4K3 activate JNK , p38 , or IKK in the absence of extracellular stimulation . By replacing the carboxy-terminal TRAF domain of TRAF2 and Q9Y4K3 with repeats of the immunophilin P62942 , we demonstrate that their effector domains are composed of their amino-terminal Zn and RING fingers . Oligomerization of the TRAF2 effector domain results in specific binding to Q13233 , a protein kinase capable of JNK , p38 , and IKK activation , and induction of P01375 and IL-1 responsive genes . P01375 also enhances the binding of native TRAF2 to Q13233 and stimulates the kinase activity of the latter . Thus , P01375 and IL-1 signaling is based on oligomerization of TRAF2 and Q9Y4K3 leading to activation of effector kinases . Modulation by cytokines of glucocorticoid action . Glucocorticoids ( GC ) are potent modulators of the inflammatory response . Their effects serve to down-regulate the inflammatory response and are mediated by genomic pathways that follow the interaction with specific receptors ( glucocorticoid receptors , GR ) . Interleukin ( IL ) -1 , P60568 , and P05231 are able to increase GC secretion by enhancing synthesis and release of P06850 and DB01285 . Cytokine effects upon steroidogenesis also occur at the adrenal level . The role of cytokines as modulators of GR has received scarce attention . IL-1 has been shown to up-regulate GR mRNA expression in hypothalamic P06850 secreting cells . On the other hand , macrophage migration inhibitory factor ( MIF ) , a T-cell product inducible by inflammatory substances including other cytokines , counterregulates GC action within the immune system . Besides immunocytes and neurons , bone cells are a sensitive target for GC and cytokines . We have found that P60568 and P05231 up-regulate remarkably the number of GR binding sites and the expression of GR mRNA in peripheral blood mononuclear cells and in osteoblast-like Saos-2 cells . Available data suggest that inflammatory cytokines have both direct and indirect effects on GC action at the target level . Autocrine-induced transcription of GR in immunocytes and/or osteoblasts could be a mechanism that restrains excess cytokine production . Factors regulating insulin-like growth factor-binding protein-3 binding , processing , and potentiation of insulin-like growth factor action . In this study , we investigated the effects of various biochemical and pharmacological agents on insulin-like growth factor ( IGF ) -binding protein-3 ( P17936 ) cell binding and action in cultured bovine fibroblasts . When cells were preincubated for 48 h with 50 nM recombinant human ( rh ) P17936 , P05019 -stimulated [3H]aminoisobutyric acid ( [125H]AIB ) uptake was enhanced 2- to 3-fold . The addition of cytoskeletal disrupting agents during the preincubation with DB05897 did not affect P17936 potentiation of P05019 action , nor did a variety of serine , aspartate , and metalloproteinase inhibitors . On the other hand , ammonium chloride and chloroquine , weak bases that neutralize the pH of acidic cell compartments , blocked P17936 potentiation of P05019 -stimulated [3H]AIB uptake . Chloroquine and ammonium chloride had no effect alone and did not inhibit P08069 binding or action in the absence of DB05897 . Bafilomycin A , a specific inhibitor of DB00171 -dependent hydrogen ion pumps , also inhibited P17936 potentiation of P05019 -stimulated [3H]AIB uptake . Competitive [125I] P05019 binding and affinity cross-linking experiments suggested structure/function changes in cell-bound P17936 that were altered in the presence of chloroquine and bafilomycin . DB01109 markedly decreased initial P17936 cell adherence , but could not promote dissociation of P17936 from cells after the 48-h preincubation . Moreover , heparin did not inhibit P17936 potentiation of P05019 action . In summary , these data indicate that P17936 undergoes specific pH-dependent structural and/or environmental modifications that mediate the enhancing effect of P17936 on P05019 action in bovine fibroblasts . They also suggest that P17936 binding to heparin-like molecules on the cell surface is not directly involved in this process . Endothelial cell transforming growth factor-β receptor activation causes tacrolimus-induced renal arteriolar hyalinosis . Arteriolar hyalinosis is a common histological finding in renal transplant recipients treated with the calcineurin inhibitor tacrolimus ; however , the pathophysiologic mechanisms remain unknown . In addition to increasing transforming growth factor ( TGF ) -β levels , tacrolimus inhibits calcineurin by binding to FK506-binding protein 12 ( P62942 ) . P62942 alone also inhibits TGF-β receptor activation . Here we tested whether tacrolimus binding to P62942 removes an inhibition of the TGF-β receptor , allowing ligand binding , ultimately leading to receptor activation and arteriolar hyalinosis . We found that specific deletion of P62942 from endothelial cells was sufficient to activate endothelial TGF-β receptors and induce renal arteriolar hyalinosis in these knockout mice , similar to that induced by tacrolimus . DB00864 -treated and knockout mice exhibited significantly increased levels of aortic TGF-β receptor activation as evidenced by Q15796 /3 phosphorylation , along with increased collagen and fibronectin expression compared to controls . Treatment of isolated mouse aortas with tacrolimus increased TGF-β receptor activation and collagen and fibronectin expression . These effects were independent of calcineurin , absent in endothelial denuded aortic rings , and could be prevented by the small molecule TGF-β receptor inhibitor SB-505124 . Thus , endothelial cell TGF-β receptor activation is sufficient to cause vascular remodeling and renal arteriolar hyalinosis . Effect of firocoxib or flunixin meglumine on recovery of ischemic-injured equine jejunum . OBJECTIVE : To determine whether treatment of horses with firocoxib affects recovery of ischemic-injured jejunum , while providing effective analgesia . ANIMALS : 18 horses . PROCEDURES : Horses ( n = 6 horses/group ) received saline ( 0.9 % NaCl ) solution ( 1 mL/50 kg , IV ) , flunixin meglumine ( 1.1 mg/kg , IV , q 12 h ) , or firocoxib ( 0.09 mg/kg , IV , q 24 h ) before 2 hours of jejunal ischemia . Horses were monitored via pain scores and received butorphanol for analgesia . After 18 hours , ischemic-injured and control mucosa were placed in Ussing chambers for measurement of transepithelial resistance and permeability to lipopolysaccharide . Histomorphometry was used to determine denuded villus surface area . Western blots for cyclooxygenase ( P36551 ) -1 and P35354 were performed . Plasma thromboxane B(2) and prostaglandin E(2) metabolite ( PGEM ) concentrations were determined . RESULTS : Pain scores did not significantly increase after surgery in horses receiving flunixin meglumine or firocoxib . Transepithelial resistance of ischemic-injured jejunum from horses treated with flunixin meglumine was significantly lower than in saline- or firocoxib-treated horses . Lipopolysaccharide permeability across ischemic-injured mucosa was significantly increased in horses treated with flunixin meglumine . Treatment did not affect epithelial restitution . P23219 was constitutively expressed and P35354 was upregulated after 2 hours of ischemia . Thromboxane B(2) concentration decreased with flunixin meglumine treatment but increased with firocoxib or saline treatment . Flunixin meglumine and firocoxib prevented an increase in PGEM concentration after surgery . CONCLUSIONS AND CLINICAL RELEVANCE : Flunixin meglumine retarded mucosal recovery in ischemic-injured jejunum , whereas firocoxib did not . Flunixin meglumine and firocoxib were effective visceral analgesics . DB09217 may be advantageous in horses recovering from ischemic intestinal injury . Bradykinin inhibits high glucose- and growth factor-induced collagen synthesis in mesangial cells through the B2-kinin receptor . Mesangial matrix expansion is an early lesion leading to glomeruloclerosis and chronic renal diseases . A beneficial effect is achieved with angiotensin I-converting enzyme inhibitors ( ACEI ) , which also favor bradykinin ( BK ) B2 receptor ( P30411 ) activation . To define the underlying mechanism , we hypothesized that P30411 activation could be a negative regulator of collagen synthesis in mesangial cells ( MC ) . We investigated the effect of BK on collagen synthesis and signaling in MC . Inflammation was evaluated by intercellular adhesion molecule-1 ( P05362 ) expression . BK inhibited collagen I and IV synthesis stimulated by high glucose , epithelial growth factor ( P01133 ) , and transforming growth factor-β ( TGF-β ) but did not alter P05362 . Inhibition of collagen synthesis was P30411 but not P46663 mediated . PKC or phosphatidylinositol 3-kinase ( PI3K ) inhibitors mimicked the BK effect . P30411 activation inhibited TGF-β- and P01133 -induced Erk1/2 , Q15796 /3 , Akt S473 , and P00533 phosphorylation . A phosphatase inhibitor prevented BK effects . The in vivo impact of P30411 on mesangial matrix expansion was assessed in streptozotocin-diabetic rodents . Deletion of P30411 increased mesangial matrix expansion and albuminuria in diabetic mice . In diabetic rats , matrix expansion and albuminuria were prevented by ACEI but not by ACEI and P30411 antagonist cotreatment . Consistently , the lowered BK content of diabetic glomeruli was restored by ACEI . In conclusion , deficient P30411 activation aggravated mesangial matrix expansion in diabetic rodents whereas P30411 activation reduced MC collagen synthesis by a mechanism targeting Erk1/2 and Akt , common pathways activated by P01133 and TGF-β . Taken together , the data support the hypothesis of an antifibrosing effect of P30411 activation . Alterations of respiratory chain complexes in sporadic pheochromocytoma . DB00139 dehydrogenase ( SDH ) has been associated with carcinogenesis in hereditary pheochromocytoma ( PC ) and paraganglioma . We investigated if a similar association applies to sporadic pheochromocytoma . No genetic alteration was found in the P21912 , Q99643 or O14521 genes of sporadic PC . However , in eight of nine sporadic PCs the SDH activity was , on average , reduced by 40 % ; moreover , the activities of the other oxidative phosphorylation ( OXPHOS ) complexes and citrate synthase were significantly lower compared to normal kidney tissue . Furthermore , immunohistochemical staining revealed a significant down-regulation of respiratory chain complexes . Since no pathogenic mutations were detected in the von Hippel-Lindau ( P40337 ) gene , we can rule out that P40337 deficiency is causing the general reduction of OXPHOS enzymes observed in the PCs investigated . In contrast to the single enzyme defects found in a subset hereditary PCs , a more generalized reduction of mitochondrial respiration seems to be present in most sporadic PCs . Strikingly , one of the nine PCs showed specific loss of complex I and a compensatory up-regulation of complexes II-V , which is a phenotype usually characteristic of oncocytic tumors . The high glucose-induced stimulation of P46663 and P30411 expression via CB(1)R activation is involved in rat podocyte apoptosis . AIMS : We examined renal kallikrein-kinin system ( KKS ) apoptosis and its related signaling pathway in rat podocytes . In addition , we studied the relationship of cannabinoid receptor 1 ( CB(1)R ) with high glucose and BK receptors . MAIN METHODS : Cell viability was determined by an MTT assay and apoptosis by DNA fragmentation assay , while gene expression was investigated by RT-PCR . Protein expression was analyzed by Western blot analysis . A chemical inhibitor or siRNA transfection was used to inhibit P46663 , P30411 , and CB(1)R signaling . KEY FINDINGS : High glucose ( 25 mM ) treatment decreased cell viability and increased DNA fragmentation . High glucose-induced DNA fragmentation and PARP and caspase-3 activations were blocked by both [ des- DB00125 (10) ] - DB06196 ( a P46663 antagonist ) and DB06196 ( a P30411 antagonist ) . High glucose also increased Akt phosphorylation , ER stress-related protein expression , and NF-κB/I-κB phosphorylation in podocytes , which was blocked by both [ des- DB00125 (10) ] -HOE 140 and DB06196 . In addition , P46663 and P30411 siRNA transfections prevented high glucose-induced Akt and NF-κB activations in rat podocytes . Moreover , AM251 ( a CB(1)R antagonist ) treatment and CB(1)R siRNA transfection blocked the high glucose-induced stimulation of BK receptor expression , Akt activation , and NF-κB activation . SIGNIFICANCE : Our study suggests that hyperglycemia induces apoptosis via the stimulation of P46663 and P30411 expression through CB(1)R activation in rat podocytes in vitro , which is associated with the development of diabetic nephropathy . Exenatide does not evoke pancreatitis and attenuates chemically induced pancreatitis in normal and diabetic rodents . The risk of developing pancreatitis is elevated in type 2 diabetes and obesity . Cases of pancreatitis have been reported in type 2 diabetes patients treated with P0C6A0 ( P43220 ) receptor agonists . To examine whether the P43220 agonist exenatide potentially induces or modulates pancreatitis , the effect of exenatide was evaluated in normal or diabetic rodents . Normal and diabetic rats received a single exenatide dose ( 0.072 , 0.24 , and 0.72 nmol/kg ) or vehicle . Diabetic ob/ob or HF- Q11206 mice were infused with exenatide ( 1.2 and 7.2 nmol·kg(-1)·day(-1) ) or vehicle for 4 wk . Post-exenatide treatment , pancreatitis was induced with caerulein ( CRN ) or sodium taurocholate ( ST ) , and changes in plasma amylase and lipase were measured . In ob/ob mice , plasma cytokines ( IL-1β , P60568 , P05231 , P13500 , IFNγ , and TNFα ) and pancreatitis-associated genes were assessed . Pancreata were weighed and examined histologically . Exenatide treatment alone did not modify plasma amylase or lipase in any models tested . Exenatide attenuated CRN-induced release of amylase and lipase in normal rats and ob/ob mice but did not modify the response to ST infusion . Plasma cytokines and pancreatic weight were unaffected by exenatide . Exenatide upregulated Reg3b but not Il6 , Ccl2 , Nfkb1 , or Vamp8 expression . Histological analysis revealed that the highest doses of exenatide decreased CRN- or ST-induced acute inflammation , vacuolation , and acinar single cell necrosis in mice and rats , respectively . Ductal cell proliferation rates were low and similar across all groups of ob/ob mice . In conclusion , exenatide did not modify plasma amylase and lipase concentrations in rodents without pancreatitis and improved chemically induced pancreatitis in normal and diabetic rodents .

MH_train_1388

MH_train_1388

MH_train_1388

interacts_with DB00472?

multiple_choice

P01308 action on H292 bronchial carcinoma cells as compared to normal bronchial epithelial cells . DB00030 may contribute to bronchial carcinoma due to P08069 activation by high local concentrations . Therefore , effects of insulin and P05019 on human bronchial carcinoma cells ( H292 ) and normal bronchial epithelium cells ( P02100 ) were studied . TGF-β was included since it also influences carcinoma progression . H292 and P02100 cells expressed both the insulin receptor and the P08069 ; in H292 cells an additional , shorter , splicing variant ( IR-A ) of the insulin receptor was present . P06213 expression was around four to five times higher in H292 than in P02100 cells at mRNA and protein levels . P01308 and TGF-β exerted contrary actions on proliferation and gene expression in H292 cells . Genes regulated by insulin , P05019 , and TGF-β were linked to inflammation , cell adhesion , muscle contraction and differentiation . P01308 and P05019 also suppressed DNA repair genes . EC(50) for insulin-induced proliferation was around 5 nM in H292 and around 30 nM P02100 cells . The EC(50) values for gene expression ranged from 9 to 90 nM in both cell types , dependent on the gene studied . In H292 cells , the proliferative response was much stronger if TGF-β was present . In P02100 cells this interaction of insulin and TGF-β was not observed , and changes in gene expression were mostly lower by at least 10-fold as compared to H292 . All in all , the insulin effects in H292 were generally much stronger than in P02100 cells and - with regard to proliferation - occurred at lower concentrations . Thus , insulin will hardly induce cancer from normal bronchial cells but may favour progression of pre-existing tumours . Autoradiographic analyses of the effects of restraint-induced stress on P08908 , P28335 and 5-HT2 receptors in the dorsal hippocampus of male and female rats . Quantitative autoradiography was used to evaluate the effects of sex and either 1 or 5 daily 2-hour sessions of restraint stress on binding at P08908 , P28335 and 5-HT2 receptors in the rat dorsal hippocampus . Neither sex nor restraint stress were found to have effects on binding at P28335 or 5-HT2 receptors . However , restraint stress increased binding of [3H]8-hydroxy-2-(di-n-propylamino)tetralin at P08908 receptors in the P22748 region and in the infrapyramidal dentate gyrus . In addition , levels of binding at P08908 receptors in the oriens and lacunosum moleculare layers of the P00915 region were significantly higher in female rats . Neither estradiol benzoate nor estradiol benzoate plus progesterone had effects on binding at hippocampal P08908 receptors in ovariectomized rats , making it unlikely that the sex differences were related to stages of the estrous cycle . Stress-induced levels of corticosterone ( O00230 ) were higher in females . Although O00230 levels in blood obtained during restraint decreased from session 1 to session 5 in both male and female rats , the decrease became significant in females only . Female rats also displayed higher levels of activity in the open field . Although activity in the open field was reduced in male and female rats after restraint , these decreases were not significant . Results are discussed in relation to anxiety and depression . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . Sulfated oxysterol , 25HC3S , is a potent regulator of lipid metabolism in human hepatocytes . Recently , a novel oxysterol , 5-cholesten-3beta , 25-diol 3-sulfate ( 25HC3S ) was identified in primary rat hepatocytes following overexpression of the cholesterol transport protein , StarD1 . This oxysterol was also detected in human liver nuclei . In the present study , 25HC3S was chemically synthesized . Addition of 25HC3S ( 6 microM ) to human hepatocytes markedly inhibited cholesterol biosynthesis . Quantitative RT-PCR and Western blot analysis showed that 25HC3S markedly decreased P04035 mRNA and protein levels . Coincidently , 25HC3S inhibited the activation of sterol regulatory element binding proteins ( SREBPs ) , suggesting that inhibition of cholesterol biosynthesis occurred via blocking P36956 activation , and subsequently by inhibiting the expression of HMG DB01992 reductase . 25HC3S also decreased P36956 mRNA levels and inhibited the expression of target genes encoding acetyl DB01992 carboxylase-1 ( ACC-1 ) and fatty acid synthase ( FAS ) . In contrast , 25-hydroxycholesterol increased P36956 and FAS mRNA levels in primary human hepatocytes . The results imply that 25HC3S is a potent regulator of SREBP mediated lipid metabolism . Role of monoamine oxidases in the exaggerated 5-hydroxytryptamine-induced tension development of human isolated preeclamptic umbilical artery . We investigated the role(s) of monoamine oxidases ( MAOs ) on the altered 5-hydroxytryptamine ( 5-HT , serotonin ) -induced tension development of the isolated umbilical artery of preeclamptic pregnancy of Chinese women . An enhanced 5-HT-induced tension development of the umbilical artery of preeclamptic pregnancy was observed when compared with that of normal pregnancy . The enhanced component of 5-HT-induced tension development was eradicated by clorgyline ( a P21397 inhibitor ) . Blockade of P29474 ( endothelial isoform nitric oxide synthase ) ( N(omega)-nitro-L-arginine methyl ester ) , 5-HT transporter ( citalopram ) , 5-HT receptor subtypes ( 5HT2B , SB 204741 ; P28335 , RS 102221 ; P34969 , SB 269970 ) , and endothelium denudation of the umbilical artery of normal pregnancy mimicked the enhanced 5-HT-induced tension development as observed in the preeclamptic tissues . In contrast , no apparent changes in 5-HT-induced tension development of the umbilical artery of preeclamptic pregnancy were observed with the same pharmacological manipulations . A decreased protein expression levels of P21397 and P29474 ( no P35228 and P27338 expression was detected ) and no change in caveolin-1 and 5-HT transporter expression were demonstrated in the umbilical artery ( endothelium intact ) lysate of preeclamptic pregnancy , compared to that of the umbilical artery of normal pregnancy . Thus , in the umbilical artery of preeclamptic pregnancy , a decrease of P21397 and P29474 protein expression levels are probably associated with , or responsible for , the exaggerated 5-HT-induced tension development . Structures of murine carbonic anhydrase IV and human carbonic anhydrase II complexed with brinzolamide : molecular basis of isozyme-drug discrimination . P22748 ( CAIV ) is a membrane-associated enzyme anchored to plasma membrane surfaces by a phosphatidylinositol glycan linkage . We have determined the 2.8-angstroms resolution crystal structure of a truncated , soluble form of recombinant murine CAIV . We have also determined the structure of its complex with a drug used for glaucoma therapy , the sulfonamide inhibitor brinzolamide ( DB01194 ) . The overall structure of murine CAIV is generally similar to that of human CAIV ; however , some local structural differences are found in the active site resulting from amino acid sequence differences in the " 130 's segment " and the residue-63 loop ( these may affect the nearby catalytic proton shuttle , DB00117 -64 ) . Similar to human CAIV , the C-terminus of murine CAIV is surrounded by a substantial electropositive surface potential that may stabilize the interaction with the phospholipid membrane . Binding interactions observed for brinzolamide rationalize the generally weaker affinity of inhibitors used in glaucoma therapy toward CAIV compared with CAII . Regulatory regions of growth-related genes can activate an exogenous gene of the alpha-fetoprotein promoter to a comparable degree in human hepatocellular carcinoma cells . We examined the transcriptional activation by the regulatory regions of the midkine ( MK ) , survivin ( Q09428 ) , cyclooxygenase-2 ( P35354 ) , telomerase reverse transcriptase ( O14746 ) and alpha-fetoprotein ( AFP ) genes in human hepatocellular carcinoma cells . Luciferase assays showed that the Q09428 regulatory region exhibited the greatest activity and that the MK regulatory region activated the reporter gene better than the enhancer-linked AFP promoter even in high-AFP-producing cells . The P35354 and O14746 regulatory regions also activated the reporter gene better than the AFP enhancer/promoter in intermediate-AFP-producing cells . Combination of the regulatory regions arranged in tandem modulated their transcriptional activities , depending on the arrangement of the promoters and cells examined . These data suggested that the regulatory regions of the growth-related genes could be useful to activate a therapeutic gene in hepatocellular carcinoma cells irrespective of the amounts of AFP production but combinatory use of the promoter regions could not always contribute to enhanced activity . Retinoic acid-receptor activation of P07988 gene transcription in respiratory epithelial cells . Retinoids are known to play important roles in organ development of the lung . Retinoids exert their activity by modulating the expression of numerous genes , generally influencing gene transcription , in target cells . In the present work , the mechanism by which retinoic acid ( RA ) regulates surfactant protein ( SP ) B expression was assessed in vitro . RA ( 9- DB00982 ) enhanced P07988 mRNA in pulmonary adenocarcinoma cells ( H441 cells ) and increased transcriptional activity of the P07988 promoter in both H441 and mouse lung epithelial cells ( MLE-15 ) . Cotransfection of H441 cells with retinoid nuclear receptor ( RAR ) -alpha , -beta , and -gamma and retinoid X receptor ( RXR ) -gamma further increased the response of the P07988 promoter to RA . Treatment of H441 cells with RA increased immunostaining for the P07988 proprotein and increased the number of cells in which the P07988 proprotein was detected . An RA responsive element mediating RA stimulation of the human P07988 promoter was identified . P10276 and -gamma and RXR-alpha but not P10826 or RXR-beta and -gamma were detected by immunohistochemical analysis of H441 cells . RA , by activating RAR activity , stimulated the transcription and synthesis of P07988 in pulmonary adenocarcinoma cells . Vitamin D analogues . The plethora of actions attributed to 1,25(OH)2D3 throughout the body have suggested potential therapeutic applications for the treatment of hyperproliferative diseases , immune dysfunction , endocrine disorders , and metabolic bone disease . However , the potent calcemic activity of the natural vitamin D hormone has precluded its use in most cases . New vitamin D analogues are under development that display greater specificity , in most cases , by retaining the therapeutic properties of 1,25(OH)2D3 , but with lower calcemic activity . Two analogues have been approved for use in patients : calcipotriol ( DB02300 from Leo Pharmaceuticals , Copenhagen , Denmark ) for the treatment of psoriasis ; and 19-nor-1,25(OH)2D2 ( DB00910 from Abbott Laboratories , Abbott Park , IL ) for secondary hyperparathyroidism . Many others analogues are currently being tested in preclinical and clinical trials for the treatment of various types of cancer and osteoporosis , and for immunosuppression . The selectivity of the analogues can be attributed to the combined interactions with four proteins : the vitamin D receptor ( P11473 ) , the serum vitamin D binding protein ( DBP ) , the vitamin D-24-hydroxylase and to a newly described membrane receptor . Low DBP affinity has been shown to be responsible for the reduced calcemic actions of calcipotriol and 22-oxacalcitriol ( O75051 ) , which is being tested for secondary hyperparathyroidism . However , the low calcemic activity of other analogues , including 19-nor-1,25(OH)2D2 , involves other , as yet undefined , mechanisms . Understanding of the molecular basis for the selectivity of the vitamin D analogues will allow the design of more effective and safer vitamin D compounds for the treatment of a wide range of clinical disorders . Structural and biochemical studies of botulinum neurotoxin serotype C1 light chain protease : implications for dual substrate specificity . Clostridial neurotoxins are the causative agents of the neuroparalytic disease botulism and tetanus . They block neurotransmitter release through specific proteolysis of one of the three soluble N-ethylmaleimide-sensitive-factor attachment protein receptors ( SNAREs ) P60880 , syntaxin , and synaptobrevin , which constitute part of the synaptic vesicle fusion machinery . The catalytic component of the clostridial neurotoxins is their light chain ( LC ) , a DB01593 endopeptidase . There are seven structurally and functionally related botulinum neurotoxins ( BoNTs ) , termed serotype A to G , and tetanus neurotoxin ( TeNT ) . Each of them exhibits unique specificity for their target SNAREs and peptide bond(s) they cleave . The mechanisms of action for substrate recognition and target cleavage are largely unknown . Here , we report structural and biochemical studies of BoNT/C1-LC , which is unique among BoNTs in that it exhibits dual specificity toward both syntaxin and P60880 . A distinct pocket ( S1 ' ) near the active site likely achieves the correct register for the cleavage site by only allowing Ala as the P1 ' residue for both P60880 and syntaxin . Mutations of this P60880 residue dramatically reduce enzymatic activity . The remote alpha-exosite that was previously identified in the complex of DB00083 -LC and P60880 is structurally conserved in BoNT/C1 . However , mutagenesis experiments show that the alpha-exosite of BoNT/C1 plays a less stringent role in substrate discrimination in comparison to that of DB00083 , which could account for its dual substrate specificity . Isolation , partial purification and characterization of nuclear retinoic acid receptors from chick skin . Nuclear receptors ( RARs ) for retinoic acid ( RA ) are considered to be the ultimate mediators of the action of RA in the control of cell differentiation and inhibition of tumorigenesis . We have isolated and partially purified and characterized RAR from a RA-responsive tissue , chick embryo skin . The purification steps included Affi-Gel blue chromatography , ultrafiltration , size exclusion chromatography , and preparative isoelectric focusing . The electrofocusing of RAR-[3H]RA complex in ampholines ( pH 3-10 ) revealed that the receptors have an isoelectric pH of 7.5 . Whereas pronase-digested the RAR-[3H]RA complex completely , DNase showed 20-35 % and RNase showed negligible digestive action on the complex . The ligand binding to RAR was completely inhibited by a mercury compound . P10276 - and P10826 -specific antibodies , on Western blot analysis , immunoreacted with a protein having a molecular weight of 50,000 , presumably RAR . Binding affinity studies revealed that biologically active analogs of RA with a free COOH group ( e.g. , 13- DB00982 , RO-13-7410 , Ch 55 , and DB04942 ) showed , like RA , high binding affinity for RAR , whereas biologically ineffective analogs of RA ( e.g. , furyl and pyridyl ) were poor binders . Other groups of retinoids , in which the COOH group was either lacking or blocked , did not bind to RAR whether or not they were biologically active . Support for association between ADHD and two candidate genes : NET1 and P21728 . Attention deficit hyperactivity disorder ( ADHD ) is a common , multifactorial disorder with significant genetic contribution . Multiple candidate genes have been studied in ADHD , including the norepinephrine transporter ( NET1 ) and dopamine D1 receptor ( P21728 ) . NET1 is implicated in ADHD because of the efficacy of atomoxetine , a selective noradrenergic reuptake inhibitor , in the treatment of ADHD . P21728 is primarily implicated through mouse models of ADHD . DNA from 163 ADHD probands , 192 parents , and 129 healthy controls was used to investigate possible associations between ADHD and polymorphisms in 12 previously studied candidate genes ( P28222 , 5- Q13049 , P28335 , P08913 , P43681 , P21964 , Q01959 , P21728 , P21917 , P21918 , NET1 , and P60880 ) . Analyses included case-control and family-based methods , and dimensional measures of behavior , cognition , and anatomic brain magnetic resonance imaging ( Q9BWK5 ) . Of the 12 genes examined , two showed a significant association with ADHD . Transmission disequilibrium test ( P04053 ) analysis revealed significant association of two NET1 single nucleotide polymorphisms ( SNPs ) with ADHD ( P < or = 0.009 ) ; case-control analysis revealed significant association of two P21728 SNPs with ADHD ( P < or = 0.008 ) . No behavioral , cognitive , or brain Q9BWK5 volume measurement significantly differed across NET1 or P21728 genotypes at an alpha of 0.01 . This study provides support for an association between ADHD and polymorphisms in both NET1 and P21728 ; polymorphisms in ten other candidate genes were not associated with ADHD . Because family-based and case-control methods gave divergent results , both should be used in genetic studies of ADHD . Docosahexaenoic acid inhibits insulin-induced activation of sterol regulatory-element binding protein 1 and cyclooxygenase-2 expression through upregulation of Q96EB6 in human colon epithelial cells . Multiple lines of compelling evidence from clinical and population-based studies support that hyperinsulinemia often accompanying obesity-associated insulin insensitivity promotes colon carcinogenesis . P01308 can acetylate , thereby activating sterol regulator element-binding protein 1 ( P36956 ) , a prime transcription factor responsible for expression of genes involved in lipogenesis . Moreover , P36956 upregulates cyclooxygenase-2 ( P35354 ) , a key player in inflammatory signaling . Docosahexaenoic acid ( DB01708 ) , a representative omega-3 polyunsaturated fatty acid , has been known to negatively regulate P36956 , but the underlying molecular mechanism is not fully clarified yet . This prompted us to investigate whether DB01708 could inhibit insulin-induced activation of P36956 and P35354 expression in the context of its potential protective effect on obesity-induced inflammation and carcinogenesis . Q96EB6 , a NAD(+)-dependent histone/non-histone protein deacetylase , has been reported to inhibit intracellular signaling mediated by P36956 through deacetylation of this transcription factor . We found that DB01708 induced Q96EB6 expression in CCD841CoN human colon epithelial cells . DB01708 abrogated insulin-induced acetylation as well as expression of P36956 and P35354 upregulation . P01308 -induced stimulation of CCD841CoN cell migration was also inhibited by DB01708 . These effects mediated by DB01708 were attenuated by pharmacologic inhibition of Q96EB6 . Hyperinsulinemia or insulin resistance is considered to be associated with obesity-associated inflammation . Genetically obese ( ob/ob ) mice showed higher colonic expression levels of both P36956 and P35354 than did normal lean mice . Likewise , expression of P36956 and P35354 was elevated in human colon tumor specimens compared with surrounding normal tissues . In conclusion , DB01708 may protect against obesity-associated inflammation and colon carcinogenesis by suppressing insulin-induced activation of P36956 and expression of P35354 through up-regulation of Q96EB6 . Use of a cyclo-oxygenase 2 inhibitor for prophylaxis of cystoid macular oedema following cataract surgery : a randomized placebo-controlled trial . BACKGROUND : To assess the efficacy of Celecoxib , a cyclo-oxygenase 2 ( P35354 ) inhibitor , as prophylaxis for cystoid macular oedema after routine cataract surgery . METHODS : A prospective , randomized , double-blind placebo-controlled trial of 69 hospital patients undergoing cataract surgery . Celecoxib 200 mg twice daily or placebo was given immediately after surgery for 14 days . Optical coherence tomography was used to quantify macular thickness before surgery and on day 1 , week 2 and week 6 after surgery . RESULTS : Sixty-nine patients were enrolled , of which 33 received placebo and 36 received active drug . Clinically apparent cystoid macular oedema occurred in four of the treatment group and two of the placebo group ( P = 0.68 ) . No difference in best-corrected visual acuity was seen at 6 weeks ( P = 0.37 ) . Covariate analysis of the results at 2 weeks and 6 weeks showed a macular thickness of 3 % less in the treatment group compared with placebo ( P = 0.050 ) . CONCLUSION : Celecoxib may decrease macular thickening following routine cataract surgery at 2 and 6 weeks after surgery as measured by Stratus O75051 III . No difference in best-corrected visual acuity or clinically apparent cystoid macular oedema was seen . Further investigation of P35354 inhibitors in a larger prospective randomized trial is required . Induction of retinoic acid receptor-beta suppresses cyclooxygenase-2 expression in esophageal cancer cells . Since retinoic acid receptor ( RAR ) -beta mRNA is frequently lost during esophageal carcinogenesis and esophageal cancer cells that do not express P10826 are resistant to retinoic acid ( RA ) , we stably transfected P10826 expression vector into an esophageal cancer cell line TE-8 and an antisense P10826 into TE-3 cells . Transfection of P10826 decreased cell growth and colony formation and induced apoptosis in TE-8 cells . Antisense P10826 -transfected TE-3 cells had a shorter doubling time and became resistant to RA . Induction of P10826 decreased P35354 expression in P10826 transfected TE-8 cells , whereas antisense P10826 transfected TE-3 cells increased P35354 expression . The inhibitory effect of P10826 on P35354 expression was further enhanced in the presence of RA , which was blocked by an RAR antagonist . The synthetic retinoid N-(4-hydroxyphenyl)retinamide , which does not bind effectively to P10826 , had no effect on P35354 suppression . Furthermore , RA blocked bile acid-induced P35354 expression and prostaglandin E(2) production only in the P10826 positive cells . Our data demonstrated that anticancer effect of P10826 may be related to its ability to suppress P35354 expression and support that the loss of P10826 expression may contribute to esophageal carcinogenesis . P01308 signaling inhibits the P28335 receptor in choroid plexus via Q96HU1 kinase . BACKGROUND : G protein-coupled receptors ( GPCRs ) interact with heterotrimeric GTP-binding proteins ( G proteins ) to modulate acute changes in intracellular messenger levels and ion channel activity . In contrast , long-term changes in cellular growth , proliferation and differentiation are often mediated by tyrosine kinase receptors and certain GPCRs by activation of mitogen-activated protein ( Q96HU1 ) kinases . Complex interactions occur between these signaling pathways , but the specific mechanisms of such regulatory events are not well-understood . In particular it is not clear whether GPCRs are modulated by tyrosine kinase receptor- Q96HU1 kinase pathways . RESULTS : Here we describe tyrosine kinase receptor regulation of a GPCR via Q96HU1 kinase . P01308 reduced the activity of the P28335 receptor in choroid plexus cells which was blocked by the Q96HU1 kinase kinase ( MEK ) inhibitor , PD 098059 . We demonstrate that the inhibitory effect of insulin and insulin-like growth factor type 1 ( DB01277 ) on the P28335 receptor is dependent on tyrosine kinase , DB01367 and Q96HU1 kinase . The effect may be receptor-specific : insulin had no effect on another GPCR that shares the same G protein signaling pathway as the P28335 receptor . This effect is also direct : activated Q96HU1 kinase mimicked the effect of insulin , and removing a putative Q96HU1 kinase site from the P28335 receptor abolished the effect of insulin . CONCLUSION : These results show that insulin signaling can inhibit P28335 receptor activity and suggest that Q96HU1 kinase may play a direct role in regulating the function of a specific GPCR . Atherosclerosis and the Glu298Asp polymorphism of the P29474 gene in white patients with end-stage renal disease . BACKGROUND : We investigated whether the P29474 G/T polymorphism ( Glu298Asp variant ) is linked to the severity of carotid atherosclerosis and whether it is independent of asymmetric dimethylarginine ( DB01686 ) in determining vascular damage in patients with end-stage renal disease ( ESRD ) . METHODS : The P29474 polymorphism , DB01686 , carotid intima-media thickness ( IMT ) , and carotid artery ( CCA ) wall-to-lumen ratio ( an indicator of arterial remodeling ) were determined/measured in 131 patients with ESRD . RESULTS : Both in the co-dominant and dominant model approach , IMT as well as CCA wall-to-lumen ratio were directly related to the T allele ( P < or = .009 ) and these relationships held true in multiple linear regression analyses including DB01686 and traditional and emerging risk factors . The relationship between P29474 genotypes and CCA wall-to-lumen ratio was further analyzed by a categorical approach and in a multiple logistic regression analysis , the odds ratio ( OR ) of increased CCA wall-to-lumen ratio was strongly associated to the T allele ( codominant model : GG , OR = 1 ; GT , OR = 2.1 ; TT , OR = 8.2 ; P for trend = .01 ; dominant model : GG , OR = 1 ; GT and TT , OR = 2.7 ; P = .02 ) . CONCLUSIONS : The T allele of P29474 gene is an independent predictor of intimal lesions and vascular remodeling and it is associated with the severity of atherosclerosis independently of DB01686 . Antiallodynic effect of etidronate , a bisphosphonate , in rats with adjuvant-induced arthritis : involvement of DB00171 -sensitive K+ channels . Bisphosphonates , pyrophosphate analogues , known as inhibitors of bone resorption , appear to cause analgesia in certain clinical painful situations . To detect clinically relevant analgesic property of etidronate , a non-aminobisphosphonate , we examined and characterized its antiallodynic effect in the rat with adjuvant-induced arthritis , in comparison with alendronate , an aminobisphosphonate , as determined by the von Frey test . Repeated systemic administration of etidronate at 10-40 mg/kg/day suppressed the adjuvant-induced mechanical allodynia in rat hindpaw , an effect reaching a plateau in approximately 10 days . Systemic or intraplantar ( i.pl. ) administration of DB00171 -sensitive K+ ( K+ DB00171 ) channel inhibitors , glibenclamide and/or tolbutamide , completely reversed the antiallodynic effect of etidronate within 1h in the arthritic rats , without affecting the nociceptive scores in naïve or arthritic animals that had not received etidronate . DB00630 , administered repeatedly , also revealed similar glibenclamide-reversible antiallodynic effect . In contrast , the antiallodynic effect of repeated systemic indomethacin was resistant to i.pl. glibenclamide in the arthritic rats . Repeated administration of etidronate or alendronate only slightly attenuated the adjuvant-evoked hindpaw edema . Among K+ DB00171 channel subunits , mRNAs for Kir6.1 , Q09428 , SUR2A and SUR2B were abundant in rat dorsal root ganglia , while Kir6.2 mRNA was poor . Our data demonstrate that repeated etidronate as well as alendronate exhibits antiallodynic activity in arthritic rats , which might be clinically relevant , and suggest involvement of K+ DB00171 channels in the underlying mechanisms .

MH_train_1389

MH_train_1389

MH_train_1389

interacts_with DB00502?

multiple_choice

P01308 like growth factor-1 selectively regulates the expression of matrix metalloproteinase-2 in malignant H-ras transformed cells . The present study demonstrates alterations in the regulation of matrix metalloproteinase-2 ( P08253 ) expression in response to insulin like growth factor-1 ( DB01277 ) in a H-ras transformed cell line , P01024 , which is capable of metastasis formation . These changes in P08253 expression in response to DB01277 treatment did not occur in either non-transformed parental 10 T 1/2 cells or in H-ras transformed cells ( Q13224 cells ) which are capable of benign tumour formation . DB01277 treatment of P01024 cells resulted in increased expression of P08253 gelatinolytic activity and increased expression of P08253 mRNA levels . The DB01277 mediated alterations in P08253 mRNA levels were dependent upon de novo protein synthesis and independent of transcriptional events , but dependent upon post-transcriptional regulatory events . Most notably , DB01277 can regulate P08253 mRNA expression in P01024 cells through a mechanism involving P08253 message stabilization . This study demonstrates aspects of the temporal regulatory mechanisms of P08253 expression in response to insulin-like growth factor-1 in a H-ras transformed fibrosarcoma cell line capable of metastasis formation and thereby , provides further insight into the altered growth regulatory program associated with H-ras mediated cellular transformation and malignant progression . Catalog of 178 variations in the Japanese population among eight human genes encoding G protein-coupled receptors ( GPCRs ) . We screened DNAs from 48 Japanese individuals for single-nucleotide polymorphisms ( SNPs ) in eight genes encoding G protein-coupled receptors ( GPCRs ) by directly sequencing the entire relevant genomic regions except for repetitive-sequence elements . This approach identified 147 SNPs and 31 insertion/deletion polymorphisms among the eight GPCR genes . On average , we identified one SNP in every 584 nucleotides . Of the 147 SNPs , 69 were identified in P30556 , 12 in P50052 , nine in P35414 , 20 in P37288 , nine in P30518 , 16 in P21728 , six in P08514 , and six in P43119 . Twenty-one SNPs were located in 5' flanking regions , 76 in introns , 32 in exons , and 18 in 3' flanking regions . These variants should contribute to investigations of possible correlations between genotypes and phenotypes as regards susceptibility to disease or responsiveness to drug therapy . Functional identification of NR2 subunits contributing to DB01221 receptors on DB05875 receptor-expressing dorsal horn neurons . DB01221 receptors are important elements in pain signaling in the spinal cord dorsal horn . They are heterotetramers typically composed of two Q9UHB4 and two of four NR2 subunits : Q12879 -2D . Mice lacking specific NR2 subunits show deficits in pain transmission yet subunit location in the spinal cord remains unclear . We have combined electrophysiological and pharmacological approaches to investigate the composition of functional DB01221 receptors expressed by lamina I , DB05875 receptor-expressing ( P25103 + ) neurons , as well as P25103 - neurons . Under low Mg2+ conditions ( 100 microM ) , the conductance of DB01221 receptors at -90 mV ( g ( -90 mV ) ) with Q12879 or Q13224 subunits ( Q12879 /B ) is low compared to conductance measured at the membrane potential where the inward current is maximal or maximal inward current ( MIC ) ( ratio of approximately 0.07 calculated from Kuner and Schoepfer , 1996 ) . For Q14957 or O15399 subunits ( Q14957 /D ) , the ratio is higher ( ratio approximately 0.4 ) . P25103 + and P25103 - neurons express DB01221 receptors that give ratios approximately 0.28 and 0.16 , respectively , suggesting both types of subunits are present in both populations of neurons , with P25103 + neurons expressing a higher percentage of Q14957 /D type DB01221 receptors . This was confirmed using EAB318 , an Q12879 /B preferring antagonist , and UBP141 , a mildly selective Q14957 /D antagonist to increase and decrease the g ( -90 mV ) /g(MIC) ratios in both subpopulations of neurons . Neuroendocrinology of the pituitary gland . The hypophysial-portal chemotransmitter hypothesis of control of the anterior pituitary was first set forth in the 1940s on the basis of physiological studies of the effects of lesions of the hypothalamus , and of section of the pituitary stalk on pituitary function . Morphological demonstration of specific neuropeptide pathways in the hypothalamus , which project to the median eminence , and the chemical identification of releasing hormones in the hypothalamus have fully established this theory . Specific neuropeptides have been isolated which stimulate the secretion of DB01285 ( CRF , corticotrophin releasing hormone ) , DB00024 ( TRH , thyrotropin releasing hormone ) , GH ( P01286 , growth hormone releasing hormone ) , and the gonadotropins ( P01148 , luteinizing hormone releasing hormone ; DB00644 , gonadotropin releasing hormone ) . P01236 secretion is regulated by both an inhibitory hormone ( dopamine ) , and by one or more releasing factors . A factor inhibitory to GH and DB00024 secretion has also been identified . All factors except for the prolactin inhibitory hormone ( which is a biogenic amine ) are peptides , all synthesized as part of large prohormones . These substances have all been introduced into medical and veterinary practice where they are useful for regulation of pituitary abnormalities , and study of normal physiology . Isoenzyme-specific cyclooxygenase inhibitors : a whole cell assay system using the human erythroleukemic cell line HEL and the human monocytic cell line Mono Mac 6 . NSAIDs inhibit the conversion of arachidonic acid into DB03866 and Prostaglandin H2 which is catalyzed by the enzyme cyclooxygenase ( P36551 ) . Two genetically distinct isoforms have been discovered , P23219 and P35354 . While P23219 is thought to account for homeostatic amounts of eicosanoids , P35354 is induced during inflammation leading to pathologic amounts of eicosanoids . Since NSAIDs inhibit both P36551 isoforms , antiinflammatory drug research has refocused to discovering P35354 inhibitors that do not inhibit P23219 . For this purpose , we have developed a whole cell assay system using the human erythroleukemic cell line HEL as a source for P23219 and the human monocytic cell line Mono Mac 6 as a source for P35354 . Mono Mac 6 cells express high amounts of P35354 upon stimulation with lipopolysaccharide ( LPS ) in the absence of any detectable P23219 protein . On the other hand , we find HEL cells to naturally express P23219 protein , but not P35354 . Testing of a panel of NSAIDs as well as some P35354 specific inhibitors showed that this assay system is suitable for identifying compounds that selectively inhibit either P23219 or P35354 . This test system offers the advantage of assessing P23219 and P35354 inhibitors within the human species , within a similar test set-up , and circumvents the need for tedious purification of either platelets or peripheral blood monocytes . DB00502 induces neurotoxicity by the DB01221 receptor downstream signaling pathway , alternative from glutamate excitotoxicity . The DB01221 receptor is believed to be important in a wide range of nervous system functions including neuronal migration , synapse formation , learning and memory . In addition , it is involved in excitotoxic neuronal cell death that occurs in a variety of acute and chronic neurological disorders . Besides of agonist/coagonist sites , other modulator sites , including butyrophenone site may regulate the N-methyl-D-aspartate receptor . It has been shown that haloperidol , an antipsychotic neuroleptic drug , interacts with the Q13224 subunit of DB01221 receptor and inhibits DB01221 response in neuronal cells . We found that DB01221 receptor was co-immunoprecipitated by anti-Ras antibody and this complex , beside NR2 subunit of DB01221 receptor contained haloperidol-binding proteins , P29475 and Ras- P01286 . Furthermore , we have shown that haloperidol induces neurotoxicity of neuronal cells via DB01221 receptor complex , accompanied by dissociation of Ras- P01286 from membranes and activation of c-Jun-kinase . Inclusion of insulin prevented relocalization of Ras- P01286 and subsequent neuronal death . DB00502 -induced dissociation of Ras- P01286 leads to inhibition of membrane-bound form of Ras protein and changes downstream regulators activity that results in the initiation of the apoptotic processes via the mitochondrial way . Our results suggest that haloperidol induces neuronal cell death by the interaction with DB01221 receptor , but through the alternative from glutamate excitotoxicity signaling pathway . Effect of valproic acid through regulation of DB01221 receptor- P29323 signaling in sleep deprivation rats . Although the effect of mood stabilizer valproic acid ( DB00313 ) through multiple signaling pathways has been shown , its therapeutic mechanism is still largely unknown . We investigated the effect of DB00313 ( 200 mg/kg , every 12 h ) in sleep deprivation ( SD ) rats ( 72 h ) , the manic-like animal model , focusing on the N-methyl-D : -aspartic acid ( DB01221 ) receptor and signaling mediators of synaptic plasticity such as extracellular signal-regulated protein kinase ( P29323 ) , DB02527 response element-binding protein ( CREB ) , B cell chronic lymphocytic leukemia/lymphoma 2 ( P10415 ) , and brain-derived neurotrophic factor ( P23560 ) . SD reduced the expression of the Q13224 subunit of the DB01221 receptor in the frontal cortex and hippocampus but did not affect the expression of Q9UHB4 and Q12879 subunits . In comparison , DB00313 inhibited the SD-induced reduction of Q13224 expression in both brain regions . In addition , SD attenuated P29323 phosphorylation in the frontal cortex and hippocampus , whereas DB00313 prevented the attenuation . DB00313 also protected the SD-induced decrease of CREB phosphorylation , P10415 expression , and P23560 expression in the frontal cortex but not in the hippocampus . These results indicate that DB00313 could regulate DB01221 receptor- P29323 signaling in SD rats , preventing the SD-induced decrease of the expression of Q13224 subunit and the activation of P29323 signaling mediators such as P29323 , CREB , P10415 , and P23560 . Maladaptive exploratory behavior and neuropathology of the P49768 P117L Alzheimer transgenic mice . Patients with the early-onset Alzheimer 's disease P117L mutation in the presenilin-1 gene ( P49768 ) present pathological hallmarks in the hippocampus , the frontal cortex and the basal ganglia . In the present work we determined by immunohistochemistry which brain regions were injured in the transgenic P49768 P117L mice , in comparison to their littermates , the B6D2 mice . Furthermore , as these regions are involved in novelty detection , we investigated the behavior of these mice in tests for object and place novelty recognition . Limited numbers of senile plaques and neurofibrillary tangles were detected in aged P49768 P117L mice in the P00915 only , indicating that the disease is restrained to an initial neuropathological stage . Western blots showed a change in P78352 expression ( p=0.03 ) , not in Q12879 subunit , Q13224 subunit and synaptophysin expressions in the frontal cortex , suggesting specific synaptic alterations . The behavioral tests repeatedly revealed , despite a non-significant preference for object or place novelty , maladaptive exploratory behavior of the P49768 P117L mice in novel environmental conditions , not due to locomotor problems . These mice , unlike the B6D2 mice , were less inhibited to visit the center of the cages ( p=0.01 ) and they continued to move excessively in the presence of a displaced object ( p=0.021 ) . Overall , the P49768 P117L mice appear to be in an initial Alzheimer 's disease-like neuropathological stage , and they showed a lack of reaction toward novel environmental conditions . Functional methionine synthase deficiency ( cblE and cblG ) : clinical and biochemical heterogeneity . Functional methionine synthase deficiency is generally characterized by homocystinuria and hypomethioninemia in the absence of methylmalonic aciduria . Patients are divided into two classes , cblE and cblG , on the basis of complementation analysis . Presentation has usually been in the first 2 years of life , but one patient came to medical attention at age 21 years with symptoms initially diagnosed as multiple sclerosis . Common findings among 11 patients ( 4 with cblE and 7 with cblG ) have included megaloblastic anemia ( all patients ) and various neurological deficits including developmental retardation ( 10 patients ) , cerebral atrophy ( 8 patients ) , hypotonia ( 7 patients ) , EEG abnormalities ( 6 patients ) , and nystagmus ( 5 patients ) . Hypertonia , seizures , blindness , and ataxia were less frequent . All patients have responded to therapy with cobalamin with resolution of anemia and biochemical abnormalities ; neurological deficits resolved more slowly and in some cases incompletely . DB00200 has been more effective than cyanocobalamin . Fibroblasts from patients with cblE ( 5 patients ) and cblG ( 6 patients ) all showed decreased intracellular levels of methylcobalamin ( DB03614 ) and decreased incorporation of label from 5-methyltetrahydrofolate into macromolecules , suggesting decreased activity of the DB03614 -dependent enzyme methionine synthase . Q99707 specific activity in extracts of all cblE fibroblasts was normal or near-normal under standard reducing conditions ; synthase specific activity in extracts of 5 cblG patients was low but was high in a 6th patient measured in another laboratory . Thus , there is heterogeneity among patients with functional methionine synthase deficiency both in clinical presentation and in the results of biochemical studies of cultured cells . Use of P01148 antagonists in reproductive medicine . Gonadotrophin-releasing hormone ( DB00644 ) plays a key role in the secretion of gonadotrophins , follicle-stimulating hormone ( DB00094 ) and luteinizing hormone ( LH ) , which regulate steroidogenesis and folliculogenesis . Two DB00644 antagonists , DB00050 and DB06785 , deprived of histaminergic side-effects , have been introduced into ovarian stimulation protocols to prevent premature LH surges and proved their safety in clinical trials . At present , most of the published studies have not found significant differences in follicular recruitment , oocyte quality , and so on , except for a decrease in pregnancy and implantation rates in in vitro fertilization and embryo transfer ( IVF-ET ) cycles when the DB00644 antagonist rather than the agonist was used . This decrease in pregnancy rates was in relation with a necessary learning curve of the physicians . Another possibility is the impact of the DB00644 antagonist on endometrium through its P30968 ; this effect was cancelled after cryopreserved embryo transfers because the pregnancy rates were similar between DB00644 antagonist and agonist in this case . DB00644 antagonists were also interesting in poor responders and polycystic ovarian syndrome , where the agonists have not permitted to obtain the better results in IVF-ET cycles . Similarly , the DB00644 antagonists could prevent the LH surge in the intrauterine insemination cycles . ZNF804a regulates expression of the schizophrenia-associated genes Q9NQE7 , P21964 , Q07343 , and P14416 . ZNF804a was identified by a genome-wide association study ( GWAS ) in which a single nucleotide polymorphism ( SNP rs1344706 ) in ZNF804a reached genome-wide statistical significance for association with a combined diagnosis of schizophrenia ( SZ ) and bipolar disorder . Although the molecular function of ZNF804a is unknown , the amino acid sequence is predicted to contain a C2H2-type zinc-finger domain and suggests ZNF804a plays a role in DNA binding and transcription . Here , we confirm that ZNF804a directly contributes to transcriptional control by regulating the expression of several SZ associated genes and directly interacts with chromatin proximal to the promoter regions of Q9NQE7 and P21964 , the two genes we find upregulated by ZNF804a . Using immunochemistry we establish that ZNF804a is localized to the nucleus of rat neural progenitor cells in culture and in vivo . We demonstrate that expression of ZNF804a results in a significant increase in transcript levels of Q9NQE7 and P21964 , relative to GFP transfected controls , and a statistically significant decrease in transcript levels of Q07343 and P14416 . Furthermore , we show using chromatin immunoprecipitation assays ( ChIP ) that both epitope-tagged and endogenous ZNF804a directly interacts with the promoter regions of Q9NQE7 and P21964 , suggesting a direct upregulation of transcription by ZNF804a on the expression of these genes . These results are the first to confirm that ZNF804a regulates transcription levels of four SZ associated genes , and binds to chromatin proximal to promoters of two SZ genes . These results suggest a model where ZNF804a may modulate a transcriptional network of SZ associated genes . Passive smoke effects on cough and airways in young guinea pigs : role of brainstem DB05875 . Children raised with extended exposure to environmental tobacco smoke ( ETS ) experience increased cough and wheeze . This study was designed to determine whether extended ETS exposure enhances citric acid-induced cough and bronchoconstriction in young guinea pigs via a neurokinin-1 ( NK-1 ) receptor mechanism at the first central synapse of lung afferent neurons , the nucleus tractus solitarius . Guinea pigs were exposed to ETS from 1 to 6 weeks of age . At 5 weeks of age , guide cannulae were implanted bilaterally in the medial nucleus tractus solitarius at a site that produced apnea in response to the glutamate agonist D,L-homocysteic acid . At 6 weeks of age , either vehicle or a P25103 antagonist , DB05790 , was injected into the nucleus tractus solitarius of the conscious guinea pigs who were then exposed to citric acid aerosol . ETS exposure significantly enhanced citric acid-induced cough by 56 % and maximal Penh ( a measure of airway obstruction ) by 43 % , effects that were attenuated by the P25103 antagonist in the nucleus tractus solitarius . We conclude that in young guinea pigs extended exposure to ETS increases citric acid-induced cough and bronchoconstriction in part by an P25103 mechanism in the nucleus tractus solitarius . Mechanism of interaction of niflumic acid with heterologously expressed kidney CLC-K chloride channels . CLC-K Cl(-) channels belong to the CLC protein family . In kidney and inner ear , they are involved in transepithelial salt transport . Mutations in ClC-Kb lead to Bartter 's syndrome , and mutations in the associated subunit barttin produce Bartter 's syndrome and deafness . We have previously found that 3-phenyl-CPP blocks hClC-Ka and rClC- P04264 from the extracellular side in the pore entrance . Recently , we have shown that niflumic acid ( DB04552 ) , a nonsteroidal anti-inflammatory fenamate , produces biphasic behavior on human CLC-K channels that suggests the presence of two functionally different binding sites : an activating site and a blocking site . Here , we investigate in more detail the interaction of DB04552 on CLC-K channels . Mutants that altered block by 3-phenyl-2-(p-chlorophenoxy)propionic acid ( CPP ) had no effect on DB04552 block , indicating that the inhibition binding site of DB04552 is different from that of 3-phenyl-CPP and flufenamic acid . Moreover , DB04552 does not compete with extracellular Cl(-) ions , suggesting that the binding sites of DB04552 are not located deep in the pore . Differently from ClC-Ka , on the rat homologue P51800 , DB04552 has only an inhibitory effect . We developed a quantitative model to describe the complex action of DB04552 on ClC-Ka . The model predicts that ClC-Ka possesses two DB04552 binding sites : when only one site is occupied , DB04552 increases ClC-Ka currents , whereas the occupation of both binding sites leads to channel block . Early structural changes in individuals at risk of familial Alzheimer 's disease : a volumetry and magnetization transfer MR imaging study . Presenilin 1 ( P49768 ) mutation carriers provide the opportunity to asses early features of neurodegeneration in familial Alzheimer 's disease ( AD ) . Gray matter ( GM ) regional volume loss and decrease of magnetization transfer ratio ( Q99707 ) consistent with microstructural changes have been reported in sporadic AD . We performed a regional volumetric and Q99707 analysis in carriers of P49768 mutations . Six non-demented mutated P49768 carriers ( 5 with memory deficits ) and 14 healthy subjects were examined with high resolution T1-weighted images for volumetry and with P24752 * weighted images for Q99707 . Cortical GM volume and Q99707 values were derived . Compared to healthy controls , the GM volume of the left temporal and inferior parietal cortex and the Q99707 of the temporal cortex bilaterally were significantly decreased in P49768 gene carriers . In the latter , the temporal lobe Q99707 showed a trend for correlation with memory and executive function scores . Early neurodegeneration in non-demented subjects at risk for familial AD may be associated with atrophy and decreased Q99707 in the temporal cortex . DB04866 : a potent inhibitor of critical steps in angiogenesis progression . We have previously demonstrated that halofuginone , a low molecular weight quinazolinone alkaloid , is a potent inhibitor of collagen alpha1(I) and matrix metalloproteinase 2 ( P08253 ) gene expression . DB04866 also effectively suppresses tumor progression and metastasis in mice . These results together with the well-documented role of extracellular matrix ( Q13201 ) components and matrix degrading enzymes in formation of new blood vessels led us to investigate the effect of halofuginone on the angiogenic process . In a variety of experimental system , representing sequential events in the angiogenic cascade , halofuginone treatment resulted in profound inhibitory effect . Among these are the abrogation of endothelial cell P08253 expression and basement membrane invasion , capillary tube formation , and vascular sprouting , as well as deposition of subendothelial Q13201 . The most conclusive anti-angiogenic activity of halofuginone was demonstrated in vivo ( mouse corneal micropocket assay ) by showing a marked inhibition of basic fibroblast growth factor ( P09038 ) -induced neovascularization in response to systemic administration of halofuginone , either i.p. or in the diet . The ability of halofuginone to interfere with key events in neovascularization , together with its oral bioavailability and safe use as an anti-parasitic agent , make it a promising drug for further evaluation in the treatment of a wide range of diseases associated with pathological angiogenesis . Exploring schizophrenia drug-gene interactions through molecular network and pathway modeling . In this study , we retrieved 39 schizophrenia-related antipsychotic drugs from the DrugBank database . These drugs had interactions with 142 targets , whose corresponding genes were defined as drug targeted genes . To explore the complexity between these drugs and their related genes in schizophrenia , we constructed a drug-target gene network . These genes were overrepresented in several pathways including : neuroactive ligand-receptor pathways , glutamate metabolism , and glycine metabolism . Through integrating the pathway information into a drug-gene network , we revealed a few bridge genes connected the sub-networks of the drug-gene network : Q12879 , O60391 , Q14957 , Q13224 , P21728 , and P14416 . These genes encode ionotropic glutamate receptors belonging to the DB01221 receptor family and dopamine receptors . DB00502 was the only drug to directly interact with these pathways and receptors and consequently may have a unique action at the drug-gene interaction level during the treatment of schizophrenia . This study represents the first systematic investigation of drug-gene interactions in psychosis . Expression of P35354 and DB01221 receptor genes at the cochlea and midbrain in salicylate-induced tinnitus . OBJECTIVE/HYPOTHESIS : The expression of the genes for cyclooxygenase ( P36551 ) and DB01221 receptor ( NR ) has seldom been reported in tinnitus . We hypothesized that expression of P35354 and NR was altered in the cochlea and midbrain in salicylate-induced tinnitus . STUDY DESIGN : Experimental study on mice . METHODS : We evaluated the tinnitus score and mRNA expression levels of P35354 and NR subtype 2B ( Q13224 ) in the cochlea and midbrain in response to intraperitoneal injections of salicylate for 4 days . RESULTS : At day 4 of tinnitus induction , the mean weights of the whole body and midbrain did not change greatly in both control and salicylate groups . The tinnitus score was not elevated from day 1 to day 4 in the control group , but increased day by day in the salicylate group . The mRNA expression level of P35354 decreased slightly in the salicylate group in the cochlea ( 1.1 ± 0.33 vs. 1.3 ± 0.49 , P = .0752 ) and in the midbrain ( 0.9 ± 0.10 versus 1.0 ± 0.35 , P = .0489 ) . Inversely , the expression levels of the Q13224 gene increased moderately in the salicylate group in the cochlea ( 3.7 ± 0.47 versus 2.3 ± 1.13 , P < 0.0001 ) and in the midbrain ( 1.6 ± 0.64 versus 1.0 ± 0.44 , P = .0007 ) . CONCLUSIONS : Salicylate induced tinnitus and altered the expression of the P35354 and Q13224 genes in the cochlea and midbrain of mice . These findings might contribute to further understanding of pathophysiology and therapy of tinnitus . Alcohols inhibit N-methyl-D-aspartate receptors via a site exposed to the extracellular environment . N-Methyl-D-aspartate ( DB01221 ) receptors are important CNS target sites of alcohols , but the site and mechanism of action of alcohols on DB01221 receptors remains unclear . In CHO- P04264 cells transfected with Q9UHB4 / Q13224 DB01221 receptor subunits , ethanol inhibited DB01221 -activated current with an IC(50) of 138 mM . Truncation of the intracellular C-terminal domain of the Q9UHB4 subunit ( NR1T ) did not alter ethanol sensitivity when combined with the Q13224 subunit , but a similar truncation of the Q13224 subunit ( NR2BT ) slightly enhanced ethanol sensitivity of receptors formed from coexpression with either Q9UHB4 or NR1T subunits . 1-Pentanol applied externally inhibited DB01221 receptors with an IC(50) of 9.9 mM , but intracellular application of 1-pentanol ( 25 mM ) did not alter DB01221 receptor inhibition by externally applied ethanol or 1-pentanol . In addition , the amplitude of DB01221 -activated current did not decrease during the time required for 1-pentanol ( 25 mM ) to diffuse throughout the cytoplasm . DB00898 did not inhibit DB01221 receptors when bath-applied in cell-attached patches or when applied to the cytoplasmic face of inside-out membrane patches . These results appear to be best explained by an action of alcohols on the DB01221 receptor-channel protein , at a site located in a domain exposed to , or only accessible from , the extracellular environment . PDE4B5 , a novel , super-short , brain-specific DB02527 phosphodiesterase-4 variant whose isoform-specifying N-terminal region is identical to that of DB02527 phosphodiesterase-4D6 ( PDE4D6 ) . The DB02527 -specific phosphodiesterase-4 ( DB05876 ) gene family is the target of several potential selective therapeutic inhibitors . The four DB05876 genes generate several distinct protein-coding isoforms through the use of alternative promoters and 5'-coding exons . Using mouse transcripts , we identified a novel , super-short isoform of human Q07343 encoding a novel 5' terminus , which we label PDE4B5 . The protein-coding region of the novel 5' exon is conserved across vertebrates , chicken , zebrafish , and fugu . Reverse-transcription-polymerase chain reaction ( PCR ) and quantitative ( PCR ) measurements show that this isoform is brain-specific . The novel protein is 58 +/- 2 kDa ; it has DB02527 hydrolyzing enzymatic activity and is inhibited by DB05876 -selective inhibitors rolipram and cilomilast ( DB03849 ) . Confocal and subcellular fractionation analyses show that it is distributed predominantly and unevenly within the cytosol . The 16 novel N-terminal residues of PDE4B5 are identical to the 16 N-terminal residues of the super-short isoform of Q08499 ( PDE4D6 ) , which is also brain-specific . PDE4B5 is able to bind the scaffold protein Q9NRI5 , whose gene has been linked to schizophrenia . Microarray expression profiling of the DB05876 gene family shows that specific DB05876 genes are enriched in muscle and blood fractions ; however , only by monitoring the individual isoforms is the brain specificity of the super-short Q08499 and Q07343 isoforms revealed . Understanding the distinct tissue specificity of DB05876 isoforms will be important for understanding phosphodiesterase biology and opportunities for therapeutic intervention . Antiplatelet effect of phloroglucinol is related to inhibition of cyclooxygenase , reactive oxygen species , P29323 /p38 signaling and thromboxane A2 production . Platelet dysfunction is a major risk factor of cardiovascular diseases such as atherosclerosis , stroke and myocardial infarction . Many antiplatelet agents are used for prevention and treatment of these diseases . In this study , phloroglucinol ( 2.5-25 μM ) suppressed AA-induced platelet aggregation and thromboxane B(2) ( TXB(2) ) production , but not U46619-induced platelet aggregation . Phloroglucinol ( 100-250 μM ) showed little cytotoxicity to platelets . Phloroglucinol inhibited the P23219 and P35354 activities by 45-74 % and 49-72 % respectively at concentrations of 10-50 μM . At concentrations of 1 and 5 μM , phloroglucinol attenuated the AA-induced ROS production in platelets by 30 % and 53 % , with an IC(50) of 13.8 μM . Phloroglucinol also inhibited the PMA-stimulated ROS production in PMN . Preincubation of platelets by phloroglucinol ( 10-25 μM ) markedly attenuated the AA-induced P29323 and p38 phosphorylation . Intravenous administration of phloroglucinol ( 2.5 and 5 μmol/mouse ) suppressed the ex vivo AA-induced platelet aggregation by 57-71 % . Phloroglucinol administration also elevated the mice tail bleeding time . Moreover , phloroglucinol inhibited the IL-1β-induced PGE(2) production in pulp fibroblasts . These results indicate that antiplatelet and anti-inflammatory effects of phloroglucinol are related to inhibition of P36551 , ROS and TXA2 production as well as P29323 /p38 phosphorylation in platelets . Phloroglucinol further suppress PMA-induced ROS production in PMN . The antiplatelet effect of phloroglucinol was confirmed by ex vivo study . Clinically , the consumption of phloroglucinol-containing food/natural products as nutritional supplement may be helpful to cardiovascular health . Phloroglucinol has potential pharmacological use . [ Meloxicam ( DB00814 ) : a review of its pharmacological and clinical profile ] . Meloxicam ( DB00814 ) is a new nonsteroidal anti-inflammatory drug ( NSAID ) derived from enolic acid , exhibiting selectivity for cyclooxygenase ( P36551 ) -2 over P23219 . Meloxicam has shown potent anti-inflammatory and analgesic activity together with low gastrointestinal toxicity in animal models . It is a potent inhibitor not only of acute exudation in adjuvant arthritis in the rat , but also of bone and cartilage destruction . The therapeutic range of meloxicam in the rat , with regard to inhibition of adjuvant arthritis , was several times greater than that of other NSAIDs . Meloxicam in therapeutic doses was found to have no effect on bleeding time or platelet aggregation in healthy volunteers . In clinical studies , meloxicam has shown reliable efficacy against rheumatoid arthritis , osteoarthritis , lumbago ( low back pain ) , scapulohumeral periarthritis , and neck-shoulder-arm syndrome with low gastrointestinal toxicity . Blocking dopamine D2 receptors by haloperidol curtails the beneficial impact of calorie restriction on the metabolic phenotype of high-fat diet induced obese mice . Calorie restriction is the most effective way of expanding life-span and decreasing morbidity . It improves insulin sensitivity and delays the age-related loss of dopamine receptor D(2) ( P14416 ) expression in the brain . Conversely , high-fat feeding is associated with obesity , insulin resistance and a reduced number of P14416 binding sites . We hypothesised that the metabolic benefit of calorie restriction involves the preservation of appropriate P14416 transmission . The food intake of wild-type C57Bl6 male mice was restricted to 60 % of ad lib. intake while they were treated with the P14416 antagonist haloperidol or vehicle using s.c. implanted pellets . Mice with ad lib. access to food receiving vehicle treatment served as controls . All mice received high-fat food throughout the experiment . After 10 weeks , an i.p. glucose tolerance test was performed and , after 12 weeks , a hyperinsulinaemic euglycaemic clamp . Hypothalamic P14416 binding was also determined after 12 weeks of treatment . Calorie-restricted ( CR ) vehicle mice were glucose tolerant and insulin sensitive compared to ad lib . ( AL ) fed vehicle mice . CR mice treated with haloperidol were slightly heavier than vehicle treated CR mice . DB00502 completely abolished the beneficial impact of calorie restriction on glucose tolerance and partly reduced the insulin sensitivity observed in CR vehicle mice . The metabolic differences between AL and CR vehicle mice were not accompanied by alterations in hypothalamic P14416 binding . In conclusion , blocking P14416 curtails the metabolic effects of calorie restriction . Although this suggests that the dopaminergic system could be involved in the metabolic benefits of calorie restriction , restricting access to high-fat food does not increase ( hypothalamic ) P14416 binding capacity , which argues against this inference . Effects of inhaled prostacyclin analogue on chronic hypoxic pulmonary hypertension . Inhaled DB01240 has been reported to elicit pulmonary vasodilation , but whether it is also effective in treating chronic hypoxic pulmonary hypertension is still uncertain . We designed this study to address the in vivo effectiveness of inhaled DB05229 , a stable DB01240 analogue , on pulmonary vascular tone during hypoxic exposure in normoxic ( N ) and chronically hypoxic ( CH ) rats . Pulmonary vasodilation was observed by low-dose inhaled DB05229 in N rats , but not in CH rats . It was not until higher doses of DB05229 were given that pulmonary vasodilation was obtained in CH rats . When the agent was continuously administered by an intravascular route at the inhaled dose , it elicited no vasodilation in N rats . On the contrary , it elicited profound vasodilation in CH rats , although a concomitant systemic hypotension was observed . The P43119 mRNA expression was unchanged in the lungs of CH rats compared with that of N rats . We conclude that low doses of aerosolized DB05229 may reduce pulmonary vascular tone in rats without preexisting lung diseases . In contrast , when hypoxic pulmonary hypertension is present , the threshold of DB05229 inhalation was elevated to provoke pulmonary vasodilation .

MH_train_1390

MH_train_1390

MH_train_1390

interacts_with DB01109?

multiple_choice

Preliminary assessment of differential expression of candidate genes associated with atherosclerosis . Identifying susceptible genes associated with the pathogenesis of atherosclerosis ( ATH ) may contribute toward better management of this condition . This preliminary study was aimed at assessing the expression levels of 11 candidate genes , namely tumor protein ( P04637 ) , transforming growth factor , beta receptor II ( P37173 ) , cysthathionenine-beta-synthase ( P35520 ) , insulin receptor substrate 1 ( P35568 ) , lipoprotein lipase ( P06858 ) , methylenetetrahydrofolate reductase ( P42898 ) , thrombomodulin ( P07204 ) , lecithin-cholesterol acyltransferase ( P04180 ) , matrix metallopeptidase 9 ( P14780 ) , low density lipoprotein receptor ( P01130 ) , and arachidonate P09917 -activating protein ( P20292 ) genes associated with ATH . Twelve human coronary artery tissues ( HCATs ) were obtained from deceased subjects who underwent post-mortem procedures . Six atherosclerotic coronary artery tissue ( ACAT ) samples representing the cases and non-atherosclerotic coronary artery tissue ( NCAT ) samples as controls were gathered based on predetermined inclusion and exclusion criteria . Gene expression levels were assessed using the GenomeLab Genetic Analysis System ( GeXP ) . The results showed that P01130 , P04637 , and P14780 expression levels were significantly increased in ACAT compared to NCAT samples ( p < 0.05 ) . Thus , P01130 , P04637 , and P14780 genes may play important roles in the development of ATH in a Malaysian study population . A phase-1 trial of bexarotene and denileukin diftitox in patients with relapsed or refractory cutaneous T-cell lymphoma . DB00004 , a genetically engineered fusion protein combining the enzymatically active domains of diphtheria toxin and the full-length sequence for interleukin-2 ( P60568 ) , efficiently targets lymphoma cells expressing the high-affinity P60568 receptor ( IL-2R ) consisting of the alpha/p55/CD25 , beta/p75/CD122 , and gamma/ P31785 /CD132 chains . In vitro studies demonstrated that the retinoid X receptor ( RXR ) retinoid , bexarotene , at biologically relevant concentrations of 10(-6) M to 10(-8) M , upregulated both the p55 and p75 subunits of the IL-2R and enhanced 5- to 10-fold the susceptibility of T-cell leukemia cells to denileukin diftitox . To determine whether this biomodulatory effect could be recapitulated in vivo , we treated 14 patients with relapsed or refractory cutaneous T-cell lymphoma with escalating doses of bexarotene ( 75 mg/day-300 mg/day ) and denileukin diftitox ( 18 mcg/kg per day x 3 days every 21 days ) in a phase 1 trial . Overall response was 67 % ( 4 complete responses , 4 partial responses ) . Modulation of IL-2R expression was observed at or above a bexarotene dose of 150 mg/day . Four patients experienced grade 2 or 3 leukopenia , and 2 had grade 4 lymphopenia . Our results demonstrate that the combination of denileukin diftitox and bexarotene is well tolerated and that even low doses ( 150 mg/day ) of bexarotene are capable of in vivo upregulation of CD25 expression on circulating leukemia cells . Functional cutaneous lymphocyte antigen can be induced in essentially all peripheral blood T lymphocytes . The cutaneous lymphocyte-associated antigen ( DB01211 ) is a skin-homing receptor expressed on a minority of memory-type peripheral blood T ( P10721 ) lymphocytes . Induction of high-level DB01211 expression in P10721 has previously been difficult to accomplish in vitro . Here we report that constitutive DB01211 expression could be readily induced in virtually all P10721 by various polyclonal activators . There was no requirement for accessory cells or addition of other mediators except for P60568 for maintaining cell survival . Absence of serum in the culture medium was important for optimal induction of DB01211 . The number of T cells adhering to P16581 as well as tethering and shear stress resistance under hydrodynamic flow increased in correlation with the level of cell surface DB01211 expressed . Clonal analysis of DB01211 induction revealed that in serum-containing medium , which permits the majority of T cells to expand , only a minority of clones did not express DB01211 . Such T cells could be induced to highly express DB01211 within 8 days by switching from serum-containing to serum-free medium . This cell-surface phenotype change was closely associated with acquisition of P16581 ligand activity . Q11130 , which is believed to be important for the generation of the DB01211 epitope on the P16109 glycoprotein ligand-1 ( Q14242 ) backbone , was shown to be significantly increased in DB01211 -positive versus DB01211 -negative T cell populations by PCR analysis . Our findings are consistent with the idea that restriction of DB01211 expression after activation , rather than positive selection of predetermined T cell subpopulations exposed to restrictive stimulatory conditions in unique microenvironments , may be important in vivo . Inhibition of a thrombin anion-binding exosite-2 mutant by the glycosaminoglycan-dependent serpins protein C inhibitor and heparin cofactor II . Antithrombin ( P01008 ) , heparin cofactor II ( HCII ) and protein C inhibitor ( P05154 ; also named plasminogen activator inhibitor-3 ) are serine protease inhibitors ( serpins ) whose thrombin inhibition activity is accelerated in the presence of glycosaminoglycans . We compared the inhibition properties of P05154 and HCII to P01008 using R93A/R97A/R101A thrombin , an anion-binding exosite-2 ( exosite-2 ) mutant that has greatly reduced heparin-binding properties . DB01109 -enhanced P05154 inhibition of R93A/R97A/R101A thrombin was only approximately 2-fold compared to 40-fold enhancement with wild-type recombinant thrombin . P07204 ( TM ) ( with or without the chondroitin sulfate moiety ) accelerated P05154 inhibition of both wild-type and R93A/R97A/R101A thrombins . HCII achieved the same maximum activity in the presence of heparin with both wild-type and R93A/R97A/R101A thrombins ; however , the optimum heparin concentration was 20 times greater than the reaction with wild-type thrombin , indicative of a decrease in heparin affinity . Dermatan sulfate ( DSO4 ) -catalyzed HCII thrombin inhibition was unchanged in R93A/R97A/R101A thrombin compared to wild-type recombinant thrombin . These results suggest that P05154 is similar to P01008 and depends upon ternary complex formation with heparin and these specific thrombin exosite-2 residues to accelerate thrombin inhibition . In contrast , HCII does not require DB00125 (93) , DB00125 (97) and DB00125 (101) of thrombin exosite-2 and further supports the hypothesis that HCII uses an allosteric process following glycosaminoglycan binding to inhibit thrombin . [ Progress in locating the genetic causes of schizophrenia ] . Elucidation of the pathogenesis of schizophrenia is progressing rapidly . The importance of the glutamatergic system and the glutamate receptor Q14832 were shown in both genetic and pharmacological studies of the new drug DB05096 . The zinc finger domain-containing gene Q7Z570 could be identified as a new schizophrenia susceptibility gene , while large copy number variants at 1q21.1 and 15q13.3 now are seen as monogenic causes of schizophrenia . It is anticipated that the coming years will see further rapid progress in the unraveling of the causes of schizophrenia . Molecular profiling to identify molecular mechanism in esophageal cancer with familial clustering . To identify the genes and molecular functional pathways involved in esophageal cancer , we analyzed the gene expression profile of esophageal tumor tissue from patients having family history of esophageal cancer by cDNA microarray . Three hundred and fifty differentially expressed genes ( 26 up-regulated and 324 down-regulated ) were identified . Genes involved in humoral immune response ( P02776 ) , extracellular matrix organization ( P53420 ) , metabolism of xenobiotics ( P07099 ) , TGF-beta signaling ( Q15797 ) and calcium signaling pathways ( P21796 ) were down-regulated and genes involved in regulation of actin cytoskeleton ( O00401 ) , neuroactive ligand receptor interaction ( Q14832 ) , Toll-like receptor ( P08571 ) , B-cell receptor ( P13164 ) and insulin signaling pathways ( Q12778 ) were up-regulated . Validation of differential expression of subset of genes by QRT-PCR and tissue microarray in familial and non-familial cases showed no significant difference in expression of these genes in two groups suggesting familial clustering occurs as result of sharing of common environmental factors . Gene expression profiling of clinical specimens from well characterized populations that have familial clustering of cancer identified molecular mechanism associated with progression of esophageal cancer . DB01109 's anti-inflammatory effects require glucosamine 6-O-sulfation and are mediated by blockade of L- and P-selectins . DB01109 has been used clinically as an anticoagulant and antithrombotic agent for over 60 years . Here we show that the potent anti-inflammatory property of heparin results primarily from blockade of P16109 and P14151 . DB01109 and chemically modified analogs were tested as inhibitors of selectin binding to immobilized sialyl Lewis(X) and of cell adhesion to immobilized selectins or thrombin-activated endothelial cells . Compared with unfractionated heparin , the modified heparinoids had inhibitory activity in this general order : over-O-sulfated heparin > heparin > 2-O,3-O-desulfated > or = N-desulfated/N-acetylated heparin > or = carboxyl-reduced heparin > or= N-,2-O,3-O-desulfated heparin >> 6-O-desulfated heparin . The heparinoids also showed similar differences in their ability to inhibit thioglycollate-induced peritonitis and oxazolone-induced delayed-type hypersensitivity . Mice deficient in P- or L-selectins showed impaired inflammation , which could be further reduced by heparin . However , heparin had no additional effect in mice deficient in both P- and L-selectins . We conclude that ( a ) heparin 's anti-inflammatory effects are mainly mediated by blocking P- and P14151 -initiated cell adhesion ; ( b ) the sulfate groups at P13671 on the glucosamine residues play a critical role in selectin inhibition ; and ( c ) some non-anticoagulant forms of heparin retain anti-inflammatory activity . Such analogs may prove useful as therapeutically effective inhibitors of inflammation . NT-702 ( parogrelil hydrochloride , DB05505 ) , a novel and potent phosphodiesterase inhibitor , improves reduced walking distance and lowered hindlimb plantar surface temperature in a rat experimental intermittent claudication model . NT-702 ( parogrelil hydrochloride , DB05505 ) , 4-bromo-6-[3-(4-chlorophenyl)propoxy]-5-[(pyridin-3-ylmethyl)amino]pyridazin-3(2H)-one hydrochloride , a novel phosphodiesterase ( PDE ) inhibitor synthesized as a potent vasodilatory and antiplatelet agent , is being developed for the treatment of intermittent claudication ( IC ) in patients with peripheral arterial disease . We assessed the efficacy of NT-702 in an experimental IC model as compared with cilostazol and additionally investigated the pharmacological property in vitro and ex vivo . NT-702 selectively inhibited PDE3 ( IC(50)=0.179 and 0.260 nM for Q14432 and 3B ) more potently than cilostazol ( IC(50)=231 and 237 nM for Q14432 and 3B ) among recombinant human PDE1 to PDE6 . NT-702 inhibited in vitro human platelet aggregation induced by various agonists ( IC(50)=11 to 67 nM ) and phenylephrine-induced rat aortic contraction ( IC(50)=24 nM ) . Corresponding results for cilostazol were 4.1 to 17 microM and 1.0 microM , respectively . NT-702 ( 3 mg/kg or more ) significantly inhibited ex vivo rat platelet aggregation after a single oral dose . For cilostazol , 300 mg/kg was effective . In a rat femoral artery ligation model , NT-702 at 5 and 10 mg/kg repeated oral doses twice a day ( P55957 ) for 13 days significantly improved the reduced walking distance while the lowered plantar surface temperature was improved at 2.5 mg/kg and more . DB01166 also improved the walking distance and surface temperature at 300 mg/kg P55957 but significant difference was only observed for surface temperature on day 8 . These results suggest that NT-702 can be expected to have therapeutic advantage for IC . Responsiveness of the QUALID to Improved Neuropsychiatric Symptoms in Patients with Alzheimer 's Disease . BACKGROUND : This study aimed to determine whether the Quality of Life in Late-Stage Dementia ( QUALID ) scale is responsive to changes in behaviour due to therapeutic intervention . METHOD : 31 long-term care residents with moderate to severe AD and agitation/aggression entered a three-month , open-label trial of memantine 10 mg P55957 . The relationships between the QUALID and BPSD , global improvement , and cognition at baseline and endpoint , as well as the changes in these scales as a result of treatment , were examined . RESULTS : Despite a significant improvement in agitation and aggression ( NPI agitation , P13726 ,90 = 3.721 , p =.014 ; CMAI total , P13726 ,90 = 6.301 , p =.001 ) and overall behaviour ( NPI total , P13726 ,90 = 4.035 , p =.010 ) , there was no significant change in QUALID score ( t30 = -0.278 , p =.783 ) . The QUALID was correlated with NPI at baseline ( τ = 0.270 , p =.037 ) and endpoint ( τ = 0.404 , p =.002 ) , but change scores were not correlated ( τ = 0.107 , p =.412 ) . CONCLUSION : While the QUALID correlates with behavioural measures at single time points , it does not appear to correlate with changes longitudinally associated with treatment . Molecular weight and biochemical profile of a chemically modified heparin derivative , Suleparoide . Recently , a new chemically modified derivative of heparin ( Suleparoide , Syntex Laboratories Buenos Aires , Argentina ) was introduced for the prophylaxis of thrombosis and treatment of vascular disorders . This agent is claimed to contain a depolymerized , chemically modified , heparin derivative with similar biologic actions as heparan sulfate . To study the pharmacologic profile of this agent , we have defined its molecular weight distribution profile , utilizing a computerized gel permeation chromatographic system equipped with ultraviolet and refractive index detectors . Suleparoide exhibited a normal molecular distribution profile with no contaminants . It exhibited a weight average of 9.3 K DA and an apparent peak MW of 8.0 K DA . Approximately 50 % of the molecular components were < 5.0 K DA and 40 % > 5.0 K DA . The results from these studies on the mechanisms show that Suleparoide has anticoagulant activity primarily mediated through DB01109 Cofactor-II ( P05546 ) and because of its novel mechanism of action , further investigations on the biochemical profile of Suleparoide are carried out . Global clotting tests such as Activated Partial P13726 Time ( APTT ) , Heptest and Thrombin Time ( TT ) revealed a concentration dependent effect in all assays . Plasma samples supplemented with Suleparoide exhibited no significant anti-Xa and anti-IIa activities . However , in the P05546 mediated inhibitory assay for IIa , Suleparoide exhibited significant activity . In contrast , the P01008 ( DB11598 ) mediated inhibition of IIa was much weaker . [ The expression of rat brain constitutive nitric oxide synthase in NG108-15 cell ] . Q8N1N2 length cDNA of rat brain P29474 was inserted into the polylinker area of pRC/CMV with specific orientation and an eukaryocyte expression vector pCMVcNOS was obtained . The existence of P29474 gene was demonstrated by PCR amplification , using pCMVcNOS gene as the model and primers designed in accord with the internal sequence of P29474 gene . The insertion and orientation of pCMVcNOS were further verified by enzymatic cleavage . NG108-15 cells were transfected with pCMVcNOS by calcium phosphate DNA coprecipitation and lipofectin transfection . G418 resistant monoclonal cells were selected with a culture medium containing 600 micrograms/ml G418 . NOS activity of each clone was assayed by monitoring the conversion of 3H- DB00125 to 3H- DB00155 . High expression cell lines were selected through measurement of the cytosol and particulate NOS activity . Out of 42 resistant monoclonal cell lines , 3 stable high expression clones have been finally selected . The increase of expressed cytosol NOS was more obvious . The result showed that the cell lines expressing P29474 at a high level had been obtained . The design and development of pegfilgrastim ( PEG-rmetHuG- P04141 , Neulasta ) . Recombinant protein technology produces drugs for human therapy in unprecedented quantity and quality . Research is now focusing on the relationship between pharmacokinetic and pharmacodynamic properties of molecules , with the aim of engineering proteins that possess enhanced therapeutic characteristics in contrast to being used as simple replacements for the natural equivalent . The addition of a polyethylene glycol ( PEG ) moiety to filgrastim ( rmetHu- DB00099 , Neupogen ) resulted in the development of pegfilgrastim . DB00019 is a long-acting form of filgrastim that requires only once-per-cycle administration for the management of chemotherapy-induced neutropenia . The covalent attachment of PEG to the N-terminal amine group of the parent molecule was attained using site-directed reductive alkylation . Pegylation increases the size of filgrastim so that it becomes too large for renal clearance . Consequently , neutrophil-mediated clearance predominates in elimination of the drug . This extends the median serum half-life of pegfilgrastim to 42 hours , compared with between 3.5 and 3.8 hours for DB00099 , though in fact the half-life is variable , depending on the absolute neutrophil count , which in turn reflects of the ability of pegfilgrastim to sustain production of those same cells . The clearance of the molecule is thus dominated by a self-regulating mechanism . DB00019 retains the same biological activity as filgrastim , and binds to the same Q99062 , stimulating the proliferation , differentiation and activation of neutrophils . Once-per-chemotherapy cycle administration of pegfilgrastim reduces the duration of severe neutropenia as effectively as daily treatment with filgrastim . In clinical trials , patients receiving pegfilgrastim also had a lower observed incidence of febrile neutropenia than patients receiving filgrastim . DB00939 sodium is an inhibitor of both the P09917 and cyclooxygenase pathways of the arachidonic acid cascade in vitro . DB00939 sodium was compared to other nonsteroidal antiinflammatory drugs in terms of its potency to inhibit the formation of 5-HETE and LTB4 in human leukocytes and the formation of prostaglandin E2 in bovine seminal vesicles as measures of its ability to inhibit the P09917 and cyclooxygenase pathways of the arachidonic acid cascade . DB00939 sodium was about 2-4 times less potent than BW-755C in inhibiting P09917 enzyme activity and three times more potent than benoxaprofen , while naproxen , ibuprofen , and indomethacin showed IC50 greater than 100 microM . DB00939 sodium and indomethacin were the most potent inhibitors of the formation of DB00917 in bovine seminal vesicles followed by ibuprofen , naproxen , and benoxaprofen in this order . DB00939 sodium , like BW-755C , can be considered a dual inhibitor of P09917 and cyclooxygenase pathways of arachidonic acid cascade . This finding may explain in part the antiinflammatory activity of meclofenamate sodium . DB09301 glycosaminoglycans as major P16109 ligands on metastatic breast cancer cell lines . The metastatic breast cancer cell line , 4T1 , abundantly expresses the oligosaccharide sialylated Lewis x ( sLe(x) ) . SLe(x) oligosaccharide on tumor cells can be recognized by E- and P16109 , contributing to tumor metastatic process . We observed that both selectins reacted with this cell line . However , contrary to the P16581 reactivity , which was sLe(x) dependent , P16109 reactivity with this cell line was sLe(x)-independent . The sLe(x)-Neg variant of the 4T1 cell line with markedly diminished expression of sLe(x) and lack of sLe(a) , provided a unique opportunity to characterize P16109 ligands and their contribution to metastasis in the absence of overlapping selectin ligands and P16581 binding . We observed that P16109 binding was Ca(2+)-independent and sulfation-dependent . We found that P16109 reacted primarily with cell surface chondroitin sulfate ( CS ) proteoglycans , which were abundantly and stably expressed on the surface of the 4T1 cell line . P16109 binding to the 4T1 cells was inhibited by heparin and CS glycosaminoglycans ( GAGs ) . Moreover , DB01109 administration significantly inhibited experimental lung metastasis . In addition , the data suggest that surface CS GAG chains were involved in P16109 mediated adhesion of the 4T1 cells to murine platelets and human umbilical vein endothelial cells . The data suggest that CS GAGs are also the major P16109 -reactive ligands on the surface of human MDA-MET cells . The results warrant conducting clinical studies on the involvement of cell surface CS chains in breast cancer metastasis and evaluation of various CS types and their biosynthetic pathways as target for development of treatment strategies for antimetastatic therapy of this disease . Cerebrospinal fluid proteomics in children during induction for acute lymphoblastic leukemia : A pilot study . BACKGROUND : Thrombosis in patients with acute lymphocytic leukemia ( ALL ) can develop after treatment with L-asparaginase ( asp ) and is often localized to the central nervous system ( CNS ) . We hypothesize that changes in the cerebrospinal fluid ( P04141 ) proteome will occur after asp therapy and will anticipate CNS clots . METHODS : Five newly diagnosed patients , ages 1-11 years , with ALL ( n = 4 ) or lymphoblastic lymphoma ( LL ) ( n = 1 ) underwent serial lumbar punctures during induction . P04141 was depleted of abundant plasma proteins and analyzed by gel-free , label-free quantitative proteomics . RESULTS : More than 600 proteins were quantified across all P04141 samples . In four subjects , the expression of proteins involved in coagulation such as protein C Inhibitor ( P05154 ) and heparin cofactor II ( P05546 ) changed over the course of asp therapy . Antithrombin III ( P01008 ) and plasminogen ( PLMN ) levels were shown to have decreased expression over time in one child who developed a CNS thrombosis , compared to other subjects . CONCLUSIONS : P04141 proteomics is feasible and reproducible in ALL and LL . P04141 P01008 and PLMN should be further investigated as predictive markers of CNS thrombosis . Endothelial protective genes induced by statin are mimicked by Q13164 activation as triggered by a drug combination of FTI-277 and GGTI-298 . BACKGROUND : Statins are potent inhibitors of cholesterol biosynthesis and are clinically beneficial in preventing cardiovascular diseases , however , the therapeutic utility of these drugs is limited by myotoxicity . Here , we explored the mechanism of statin-mediated activation of Q13164 in the human endothelium with the goal of identifying compounds that confer endothelial protection but are nontoxic to muscle . METHODS : An Q13164 -one hybrid luciferase reporter transfected into COS-7 cells with pharmacological and molecular manipulations dissected the signaling pathway leading to statin activation of Q13164 . qRT-PCR of HUVEC cells documented the transcriptional activation of endothelial-protective genes . Lastly , morphological and cellular DB00171 analysis , and induction of atrogin-1 in C2C12 myotubes were used to assess statin-induced myopathy . RESULTS : Statin activation of Q13164 is dependent on the cellular reduction of GGPPs . Furthermore , we found that the combination of FTI-277 ( inhibitor of farnesyl transferase ) and GGTI-298 ( inhibitor of geranylgeranyl transferase I ) mimicked the statin-mediated activation of Q13164 . FTI-277 and GGTI-298 together recapitulated the beneficial effects of statins by transcriptionally upregulating anti-inflammatory mediators such as P29474 , P07204 , and Q9Y5W3 . Finally , C2C12 skeletal myotubes treated with both FTI-277 and GGTI-298 evoked less morphological and cellular changes recognized as biomarkers of statin-associated myopathy . CONCLUSIONS : Statin-induced endothelial protection and myopathy are mediated by distinct metabolic intermediates and co-inhibition of farnesyl transferase and geranylgeranyl transferase I confer endothelial protection without myopathy . GENERAL SIGNIFICANCE : The combinatorial FTI-277 and GGTI-298 drug regimen provides a promising alternative avenue for endothelial protection without myopathy . Control of phenylalanine and tyrosine metabolism by phosphorylation mechanisms . A system for the parallel determination of enzyme phosphorylation and expressed activity in rat liver cells , and its application to studies of phenylalanine hydroxylase and tyrosine aminotransferase , is described . DB00120 hydroxylase is phosphorylated by agents which stimulate cyclic AMP- and Ca2+-dependent protein kinase activity . The phosphorylation site(s) appear to be the same for both kinases . Phosphorylation is accompanied by increased metabolic flux at low , physiologically relevant , substrate concentrations . P01308 and spermine both inhibit the phosphorylation of the enzyme , possibly by increasing dephosphorylation . P17735 is phosphorylated in liver cell incubations but the rate is slow and insensitive to additions to the medium . No parallel changes in flux could be detected . Both enzymes are subject to complex regulatory mechanisms , short- and long-term . Their activities may be coordinated in vivo by control exerted at the level of the plasma membrane where both amino acids share the same transport processes . Determination of the control coefficients for the several components indicates that membrane transport may be a major limitation on flux . [ Activation of coagulation cascade in children during an idiopathic nephrotic syndrome relapse ] . The objective of this study was to assess concentrations of selected markers of coagulation in children with relapse of idiopathic nephrotic syndrome during a 6-week therapy . Study groups : 22 subjects ( 32 relapses ) -- 14 males , 8 females ( mean age 7.15 +/- 1.5 y. ) with no thrombotic complications were included into the study . All children were clinically steroid-sensitive . METHODS : Coagulation markers ( platelet count , thrombin time , APTT , INR , fibrinogen 1 + 2 fragments ( F1 + 2 ) , thrombin-antithrombin complexes ( TAT ) , serum levels of D-dimer ( DD ) , fibrin monomers ( FM ) and antithrombin activity ( P01008 ) ) were measured three times : on admission , after 2 and 6 weeks . The control group consisted of 13 healthy children . RESULTS : Serum concentration of TAT or F1 + 2 did not differ between 3 stages ( p > 0.05 ) . However , values at 0 and 2 weeks were significantly higher than in control group ( p < 0.05 ) . We found no correlation between TAT or F1 + 2 and FBG , ALB , TCH , TG levels . [ table : see text ] CONCLUSIONS : The coagulation cascade in relapse of NS was activated during first 6 weeks of therapy whereas metabolic disturbances ( low ALB , high P02675 , TCH , TG , high platelets ) normalized . It is speculative whether it was caused by active immunological process but definitely it resulted in " prothrombotic state " in P01308 patients . P01308 -like growth factor-I is a serum component stimulating growth of human neuroblastoma . Human non-autocrine neuroblastoma cells SK-N-SH and LF require serum for proliferation in vitro . We wished to determine the role of serum-borne insulin-like growth factor I ( P05019 ) as mitogen for these cells . Introduction of the monoclonal antibody alpha-IR3 against human P08069 reduced proliferation in the presence of fetal bovine serum ( FBS ) . P05019 ( 5 nM ) was as effective as FBS ( 10 % ) in stimulating proliferation . DB00071 mimicked the effects of P05019 , but at a 1000-fold higher concentration . The antibody alpha-IR3 reduced growth stimulated by P05019 more effectively than growth stimulated by insulin . Thus , proliferation of human non-autocrine neuroblastoma cells can be effectively manipulated by exogenous P05019 . [ Effect of the monophase oral contraceptive combination with 20 ug ethinyl estradiol/150 ug desogestrel on haemostasis ] . The authors examined the changes in the haemostasis during the use of the oral contraceptive combination with 20 microg ethynil estradiol/150 microg desogestrel at 35 women , a basic group , who used the oral contraceptive in the duration of 12 months and a control group ( n=35 ) , who do not use the pills . We found statistically significant increase of Antithrombin III ( P01008 ) ( p < 0.011 ) , Cofactor II of DB01109 ( HCII ) ( p < 0.001 ) , the activity of plasminogen ( p < 0.026 ) and beta2-antiplasmin ( 0.026 ) , significant decrease of P02810 ( PrC ) ( p < 0.0001 ) and of total Protein S ( TPrS ) ( p < 0.03 ) in the basic group in comparision with the control one . We do not observe significant changes in the rest of the haemostatic variables between the two groups . During the use of the oral contraceptive combination with 20 microg ethynil estradiol/150 microg desogestrel the changes in the system of the natural inhibitors are balanced by these in the system of fibrinolysis . P14416 stimulation of Na+/H+ exchange assessed by quantification of extracellular acidification . A microphysiometer was used to quantify the rate of extracellular acidification by P13671 glioma cells and L fibroblasts expressing recombinant dopamine D2 receptors . The dopamine D2 receptor agonist , quinpirole , accelerated the rate of acidification of the medium by P13671 cells expressing either the short or long form of D2 receptors , D2(415) and D2(444) , but not by wild-type cells that were not transfected with a D2 receptor cDNA . The rate of acidification increased with increasing concentrations of quinpirole up to 100 nM . Inhibition of the response by the dopamine D2 antagonist , spiperone , provided additional evidence that the enhanced extracellular acidification resulted from stimulation of D2 receptors . To test the hypothesis that D2 receptor-stimulated extracellular acidification was due to transport of protons by a Na+/H+ antiporter and reflected intracellular alkalinization , the effect of two inhibitors of Na+/H+ exchange , amiloride and methyl-isobutyl-amiloride , was determined . Both compounds inhibited quinpirole-induced extracellular acidification at concentrations that did not alter D2 receptor-mediated inhibition of adenylylcyclase or radioligand binding to D2 receptors . In addition , quinpirole-induced extracellular acidification was greatly inhibited by removal of sodium from the extracellular medium , confirming the participation of Na+/H+ exchange in the extrusion of acid . Quinpirole ( 100 nM ) also increased the rate of extracellular acidification by L cells expressing D2(415) , LZR1 cells . Treatment with pertussis toxin ( 100 ng/ml for 18 h ) had no effect on the quinpirole-induced acid extrusion by C6D2(415) and LZR1 cells , although the same pertussis toxin treatment regimen completely prevented inhibition of adenylylcyclase . We conclude that recombinant D2 receptors accelerate Na+/H+ exchange in P13671 cells and L fibroblasts by a pathway that does not involve inhibition of adenylylcyclase or pertussis toxin-sensitive G proteins . Investigation of a potential protective mechanism against heparin-induced thrombocytopenia in patients on chronic intermittent hemodialysis . BACKGROUND : DB01109 -induced thrombocytopenia ( HIT ) develops as a result of platelet ( Q02083 ) activation by anti-platelet factor 4 ( P02776 ) /heparin complex antibodies . Despite repeated exposure to heparin , patients undergoing chronic intermittent hemodialysis ( HD ) rarely develop HIT . We investigated the possibility that HD decreases/removes P02776 from Q02083 surfaces and/or plasma , thereby disfavoring immune complex formation as a mechanism of protection against HIT . MATERIALS AND METHODS : We enrolled 20 patients undergoing chronic HD at the Penn Presbyterian Medical Center . Blood samples were drawn before , during and after treatment in the presence and absence of heparin . P02776 , anti- P02776 /heparin antibody , heparin , and P16109 levels were measured . RESULTS : No patients demonstrated clinical symptoms of HIT . Q02083 surface P02776 levels decreased and plasma P02776 levels increased concurrently with the increase in plasma heparin concentration . In the absence of heparin , Q02083 surface and plasma P02776 levels were unchanged . Anti- P02776 /heparin antibodies , which were non-functional by the serotonin release assay , were detectable in 8 patients . Q02083 surface P16109 levels did not change during treatment . CONCLUSIONS : Removal of Q02083 surface and/or plasma P02776 as a mechanism of protection against HIT in patients undergoing HD is not supported by the results of our study , although the transient decrease in Q02083 surface P02776 in the presence of large amounts of heparin remains a candidate mechanism . The small sample size , single type of dialyzer membrane , and early sampling time points may have led to the inability to detect changes in P02776 levels . Future studies should explore other potential protective mechanisms . DB06016 ( RGH-188 ) , a potent D3/D2 dopamine receptor partial agonist , binds to dopamine D3 receptors in vivo and shows antipsychotic-like and procognitive effects in rodents . We investigated the in vivo effects of orally administered cariprazine ( RGH-188 ; trans-N-{4-[2-[4-(2,3-dichlorophenyl)-piperazin-1-yl]-ethyl]-cyclohexyl}-N',N'-dimethyl-urea ) , a D(3)/ P14416 partial agonist with ∼10-fold preference for the D(3) receptor . Oral bioavailability of cariprazine at a dose of 1mg/kg in rats was 52 % with peak plasma concentrations of 91ng/mL . DB06016 10mg/kg had good blood-brain barrier penetration , with a brain/plasma AUC ratio of 7.6:1 . In rats , cariprazine showed dose-dependent in vivo displacement of [ (3)H ] (+)-PHNO , a dopamine D(3) receptor-preferring radiotracer , in the D(3) receptor-rich region of cerebellar lobules 9 and 10 . Its potent inhibition of apomorphine-induced climbing in mice ( ED(50)=0.27mg/kg ) was sustained for 8h . DB06016 blocked amphetamine-induced hyperactivity ( ED(50)=0.12mg/kg ) and conditioned avoidance response ( CAR ) ( ED(50)=0.84mg/kg ) in rats , and inhibited the locomotor-stimulating effects of the noncompetitive DB01221 antagonists MK-801 ( ED(50)=0.049mg/kg ) and phencyclidine ( ED(50)=0.09mg/kg ) in mice and rats , respectively . It reduced novelty-induced motor activity of mice ( ED(50)=0.11mg/kg ) and rats ( ED(50)=0.18mg/kg ) with a maximal effect of 70 % in both species . DB06016 produced no catalepsy in rats at up to 100-fold dose of its CAR inhibitory ED(50) value . DB06016 0.02-0.08mg/kg significantly improved the learning performance of scopolamine-treated rats in a water-labyrinth learning paradigm . Though risperidone , olanzapine , and aripiprazole showed antipsychotic-like activity in many of these assays , they were less active against phencyclidine and more cataleptogenic than cariprazine , and had no significant effect in the learning task . The distinct in vivo profile of cariprazine may be due to its higher affinity and in vivo binding to D(3) receptors versus currently marketed typical and atypical antipsychotics .

MH_train_1391

MH_train_1391

MH_train_1391

interacts_with DB06779?

multiple_choice

Effects of cytokines on P15692 expression and secretion by human first trimester trophoblast cell line . PROBLEM : The mechanism through which vascular endothelial growth factor ( P15692 ) regulation occurs at the feto-maternal interface is poorly understood . The aim of this study was to investigate the effects of various cytokines on P15692 expression and secretion by trophoblast cells . METHOD OF STUDY : We investigated the effects of cytokines on P15692 expression in human first trimester trophoblast cell line by analyzing P15692 messenger RNA ( mRNA ) by reverse transcription-polymerase chain reaction and P15692 protein secretion by enzyme linked immunosorbent assay . RESULTS : The trophoblast cells expressed P15692 mRNA constitutively and the main subtypes were identified as VEGF121 and VEGF165 . When cultured in the presence of interferon ( IFN ) -gamma , interleukin ( IL ) - 1beta , tumor necrosis factor ( P01375 ) -alpha , P60568 , or P22301 , P15692 mRNA expression was found to be significantly increased by IL-1beta , P01579 and P01375 but to be unaffected by P60568 and P22301 . Moreover , P15692 secretion was most significantly increased by P01579 treatment . CONCLUSION : These results suggest that IL-1beta , P01579 , and P01375 may regulate the production of P15692 in early gestational trophoblasts . P10275 YAC transgenic mice recapitulate SBMA motor neuronopathy and implicate VEGF164 in the motor neuron degeneration . X-linked spinal and bulbar muscular atrophy ( SBMA ) is an inherited neuromuscular disorder characterized by lower motor neuron degeneration . SBMA is caused by polyglutamine repeat expansions in the androgen receptor ( AR ) . To determine the basis of AR polyglutamine neurotoxicity , we introduced human AR yeast artificial chromosomes carrying either 20 or 100 CAGs into mouse embryonic stem cells . The AR100 transgenic mice developed a late-onset , gradually progressive neuromuscular phenotype accompanied by motor neuron degeneration , indicating striking recapitulation of the human disease . We then tested the hypothesis that polyglutamine-expanded AR interferes with CREB binding protein ( CBP ) -mediated transcription of vascular endothelial growth factor ( P15692 ) and observed altered CBP-AR binding and P15692 reduction in AR100 mice . We found that mutant AR-induced death of motor neuron-like cells could be rescued by P15692 . Our results suggest that SBMA motor neuronopathy involves altered expression of P15692 , consistent with a role for P15692 as a neurotrophic/survival factor in motor neuron disease . DB00065 monotherapy for refractory psoriasis : preliminary results . Tumour necrosis factor ( P01375 ) -alpha plays an important role in the pathogenesis of psoriasis . DB00065 is an anti- P01375 chimeric monoclonal antibody , which is licensed for the treatment of rheumatoid arthritis and Crohn 's disease . Some reports have shown the efficacy of infliximab , either in monotherapy or in combination with methotrexate , for the treatment of psoriatic arthropathy and psoriasis . The efficacy and tolerability of infliximab monotherapy was evaluated in 29 patients with moderate to severe psoriasis , unresponsive to conventional treatments . Fourteen patients suffered from concomitant arthropathy . Patients received intravenous infliximab , 5mg/kg , at weeks 0 , 2 , and 6 . After this 3-dose-induction regimen , patients were followed-up at monthly intervals and retreated with a single-dose infusion in case of relapse of signs and symptoms . Clinical assessment was performed using the psoriasis area and severity index ( PASI ) to monitor psoriasis activity ; pruritus and joint pain were assessed on a scale of 0 to 3 . A marked improvement of skin lesions and subjective symptoms was noted in the majority of patients ; an excellent reduction of PASI score ( > or =75 % ) was observed in 13.8 % of cases at week 2 , 71.4 % at week 6 and 78.6 % at week 10 . During the follow-up period , some patients maintained satisfactory clinical results without requiring any additional infusions . In general , skin lesions showed a trend towards a more prolonged and sustained improvement as compared with subjective symptoms . Treatment was well tolerated and no serious adverse events occurred . Multipotent cancer stem cells derived from human malignant peritoneal mesothelioma promote tumorigenesis . During the progression of malignant peritoneal mesothelioma ( MPeM ) , tumor nodules propagate diffusely within the abdomen and tumors are characterized by distinct phenotypic sub-types . Recent studies in solid organ cancers have shown that cancer stem cells ( CSCs ) play a pivotal role in the initiation and progression of tumors . However , it is not known whether tumorigenic stem cells exist and whether they promote tumor growth in MPeM . In this study , we developed and characterized a CSC model for MPeM using stably expandable tumorigenic stem cells derived from patient tumors . We found morphologically distinct populations of CSCs that divide asymmetrically or symmetrically in MPeM in vitro cell culture . The MPeM stem cells ( MPeMSCs ) express stem cell markers c-MYC , P48681 and P35968 and in the presence of matrix components cells form colony spheres . MPeMSCs are multipotent , differentiate into neuronal , vascular and adipose progeny upon defined induction and the differentiating cells express lineage-specific markers such as Q13509 , an early neuronal marker ; P04275 , P15692 , P49767 and P10145 , endothelial markers ; and PPARγ and P15090 , adipose markers . Xenotransplantation experiments using MPeMSCs demonstrated early tumor growth compared with parental cells . Limiting dilution experiments using MPeMSCs and endothelial lineage-induced cells derived from a single MPeMSC resulted in early tumor growth in the latter group indicating that endothelial differentiation of MPeMSCs is important for MPeM tumor initiation . Our observation that the MPeM tumors contain stem cells with tumorigenic potential has important implications for understanding the cells of origin and tumor progression in MPeM and hence targeting CSCs may be a useful strategy to inhibit malignant progression . Detection and quantification of cimicoxib , a novel P35354 inhibitor , in canine plasma by HPLC with spectrofluorimetric detection : development and validation of a new methodology . DB05095 ( CX ) is a selective P35354 inhibitor recently launched on the veterinary market . No analytical method to detect CX in biological samples has been published to date . The chromatographic separation was performed with a Kinetex C18 analytical column ( 100 mm × 4.6 mm , 2.6 μm particle size ) at 25 °C . The mobile phase consisted of acetonitrile:buffer ( 10 mM AcONH4 , pH 4.5 ) ( 35:65 , v/v ) at a flow rate of 1 mL/min . Excitation and emission wavelengths were 268 and 430 nm , respectively . The extraction used 500 μL of plasma added to 100 μL of IS ( 5 μg/mL ) and 100 μL of 10 % CF3COOH , extracted with 600 μL of C2H2:Et2O ( 3:7 , v/v ) . The organic phase was evaporated and reconstituted with 200 μL of mobile phase . The CX recovery ranged from 74.5 % to 82.6 % . The limit of quantification was 25 ng mL(-1) . The chromatographic runs were specific with no interfering peaks at the retention times of the analytes , as confirmed by HPLC-mass spectrometry experiments . The other validation parameters were in agreement with the international guidelines . The method was successfully tested on two dogs treated at two dose rates . It facilitated tracking of the plasma concentration for 24h and calculation of the main pharmacokinetic parameters . In conclusion , this method ( extraction , separation and applied techniques ) is simple , effective and specific . This is the first time that a method for the quantification of CX in plasma has been reported . This technique may have applications for further pharmacokinetic studies . A synthetic dl-nordihydroguaiaretic acid ( Nordy ) , inhibits angiogenesis , invasion and proliferation of glioma stem cells within a zebrafish xenotransplantation model . The zebrafish ( Danio rerio ) and their transparent embryos represent a promising model system in cancer research . Compared with other vertebrate model systems , we had previously shown that the zebrafish model provides many advantages over mouse or chicken models to study tumor invasion , angiogenesis , and tumorigenesis . In this study , we systematically investigated the biological features of glioma stem cells ( GSCs ) in a zebrafish model , such as tumor angiogenesis , invasion , and proliferation . We demonstrated that several verified anti-angiogenic agents inhibited angiogenesis that was induced by xenografted-GSCs . We next evaluated the effects of a synthetic dl-nordihydroguaiaretic acid compound ( dl-NDGA or " Nordy " ) , which revealed anti-tumor activity against human GSCs in vitro by establishing parameters through studying its ability to suppress angiogenesis , tumor invasion , and proliferation . Furthermore , our results indicated that Nordy might inhibit GSCs invasion and proliferation through regulation of the arachidonate P09917 ( Alox-5 ) pathway . Moreover , the combination of Nordy and a P15692 inhibitor exhibited an enhanced ability to suppress angiogenesis that was induced by GSCs . By contrast , even following treatment with 50 µM Nordy , there was no discernible effect on zebrafish embryonic development . Together , these results suggested efficacy and safety of using Nordy in vivo , and further demonstrated that this model should be suitable for studying GSCs and anti- P56915 drug evaluation . Pharmacodynamics and pharmacokinetics of DB05225 , a novel inhibitor of P09917 -activating protein ( P20292 ) . The P09917 -activating protein ( P20292 ) gene and an increase in leukotriene ( LT ) production are linked to the risk of asthma , myocardial infarction , and stroke . We evaluated the pharmacodynamics , pharmacokinetics , and tolerability of 3-[3-tert-butylsulfanyl-1-[4-(6-methoxy-pyridin-3-yl)-benzyl]-5-(pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid ( DB05225 ) , a novel P20292 inhibitor , in healthy subjects . Single and multiple doses of DB05225 demonstrated dose-dependent inhibition of blood Q06643 (4) production and dose-related inhibition of urinary LTE(4) . After a single oral dose ( 50-1,000 mg ) of DB05225 , the maximum concentration ( C(max) ) and area under the curve ( AUC ) in plasma increased in a dose-dependent manner . After multiple-dose administration ( 50-1,000 mg once daily for 11 days ) , there were no significant differences in the pharmacokinetic parameters between the first and last days of treatment . DB05225 was well tolerated at all doses in both the single- and multiple-dose cohorts . Further clinical trials with DB05225 in inflammatory diseases are warranted . Vascular endothelial growth factor signaling is required for the behavioral actions of antidepressant treatment : pharmacological and cellular characterization . This study extends earlier work on the role of vascular endothelial growth factor ( P15692 ) in the actions of antidepressant treatment in two key areas . First , by determining the requirement for P15692 in the actions of a 5-HT selective reuptake inhibitor ( SSRI ) , fluoxetine in behavioral models of depression/antidepressant response ; and second , by examining the role of the P08908 receptor subtype in the regulation of P15692 , and the cellular localization of antidepressant regulation of P15692 expression . The results show that pharmacological inhibition of P15692 receptor signaling blocks the behavioral actions of fluoxetine in rats subjected to chronic unpredictable stress . Infusions of SU5416 or SU1498 , two structurally dissimilar inhibitors of P15692 -Flk-1 receptor signaling , block the antidepressant effects of fluoxetine on sucrose preference , immobility in the forced swim test , and latency to feed in the novelty suppressed feeding paradigm . We also show that activation of P08908 receptors is sufficient to induce P15692 expression and that a P08908 antagonist blocks both the increase in P15692 and behavioral effects induced by fluoxetine . Finally , double labeling studies show that chronic fluoxetine administration increases P15692 expression in both neurons and endothelial cells in the hippocampus . Taken together these studies show that P15692 is necessary for the behavioral effects of the SSRI fluoxetine , as well as norepinephrine selective reuptake inhibitor , and that these effects may be mediated by P08908 receptors located on neurons and endothelial cells . Prion protein stimulates tissue-type plasminogen activator-mediated plasmin generation via a lysine-binding site on kringle 2 . Recombinant human prion-protein ( PrP23-231 ) stimulates plasminogen activation by tissue-type plasminogen activator ( t-PA ) . The stimulatory activity is conserved in the N-terminal fragment ( PrP23-110 ) . It has further been shown by others that P04156 (c) binds to kringle-domains of plasminogen . We compared the stimulatory activity of recombinant PrP23-231 and PrP23-110 on plasminogen activation catalyzed by t-PA , urokinase ( u-PA ) , streptokinase and Desmodus salivary plasminogen activator ( DSPAalpha1 ) . As these plasminogen activators are distinct , with respect to their kringle domains we studied their binding to immobilized PrP23-110 . P00747 activation was measured in a chromogenic assay in vitro and binding studies were carried out using surface plasmon resonance technology . We found that recombinant full-length prion protein , PrP23-231 , and PrP23-110 specifically stimulate t-PA mediated plasminogen activation . Two hundred nanomoles per liter of PrP23-110 stimulated 1.8 nmol L(-1) t-PA 48-fold , 180 nmol L(-1) DB04925 (alpha1) 2.5-fold , 1.8 nmol L(-1) u-PA 1.1-fold , and 1.8 nmol L(-1) streptokinase 1.8-fold . Our data show no specific binding for streptokinase . In contrast all plasminogen activators carrying a kringle domain bound to PrP23-110 . We further studied the effect of lysine on binding to PrP23-110 and on plasminogen activation by DB04925 (alpha1) or t-PA . DB00123 decreased both the binding of t-PA to PrP23-110 and the stimulation of plasmin generation by t-PA . Both binding and plasminogen activation of DB04925 (alpha1) were not influenced by the presence of lysine . All plasminogen activators tested bearing kringle domains bind to PrP23-110 . Binding to PrP23-110 is not sufficient for stimulation of plasmin generation . Thus the lysine-binding site of kringle 2 that is unique to t-PA appears to mediate the specific stimulation of plasminogen activation by the cellular prion protein . P10275 abnormalities in identical twins with oligospermia . Clinical and biochemical studies . Identical twin brothers presented with oligospermia , small testes , normal male phenotypes , elevated serum luteinizing hormone levels , and normal or elevated serum testosterone levels . Both men had low to low-normal cytosol androgen receptor binding capacity in cultured fibroblasts from pubic skin biopsy specimens . Qualitative abnormalities of cellular androgen receptors were suggested by low-normal or low nuclear androgen uptake in fibroblasts from both brothers as well as abnormal thermolability and subnormal molybdate stabilization of androgen receptors from one brother . In vivo androgen sensitivity was assessed in one twin following administration of testosterone or the non-aromatizable androgen fluoxymesterone . DB01185 suppressed serum luteinizing hormone and serum testosterone/estradiol-binding globulin , and although testosterone suppressed both serum luteinizing hormone and serum follicle-stimulating hormone , the suppression of serum luteinizing hormone by testosterone was subnormal . Both subjects showed marked exaggeration of the serum 17-hydroxyprogesterone increase after administration of human chorionic gonadotropin , despite normal serum testosterone increases , suggesting a block in testicular 17,20-desmolase , which converts 17-hydroxyprogesterone to testosterone . These studies suggest that oligospermia and block of the enzyme 17,20-desmolase may be the earliest manifestations of androgen resistance , and the finding of the syndrome of oligospermia , normal male phenotype , and androgen receptor abnormalities in identical twins indicates a genetic etiology of this disorder . Differential in vitro sensitivity to patupilone versus paclitaxel in uterine and ovarian carcinosarcoma cell lines is linked to tubulin-beta-III expression . OBJECTIVE : To compare the in vitro sensitivity/resistance to patupilone versus paclitaxel in uterine and ovarian carcinosarcomas ( CS ) . METHODS : Five primary carcinosarcoma cell lines , two from uterine and three of ovarian origin , were evaluated for growth rate and tested for their in vitro sensitivity/resistance to patupilone versus paclitaxel by MTS assays . To identify potential mechanisms underlying the differential sensitivity/resistance to patupilone , expression levels of β-tubulin III ( Q13509 ) were determined with quantitative-real-time-polymerase-chain-reaction ( q-RT-PCR ) in primary uterine and ovarian CS cell lines and in 26 uterine and 9 ovarian CS fresh-frozen-tissues . RESULTS : No appreciable difference in sensitivity to patupilone versus paclitaxel was noted in ovarian CS cell lines , or when uterine and ovarian CS cell lines were compared in their response to paclitaxel . In contrast , uterine CS cell lines were found to be significantly more sensitive to patupilone than to paclitaxel ( P < 0.002 ) and demostrated lower IC(50s) to patupilone ( range 0.76-0.93nM ) when compared to ovarian CS ( range 1.9-3.4 nM , p < 0.05 ) . Higher levels of Q13509 were detected in uterine CS cell lines and fresh frozen tissues when compared to ovarian CS ( P < 0.05 ) . CONCLUSIONS : Uterine CS cell lines are significantly more sensitive than ovarian CS cell lines to patupilone versus paclitaxel . High expression of Q13509 is associated with sensitivity to patupilone in primary CS cell lines and may act as a genetic marker to predict chemotherapy efficacy . DB03010 may represent a promising drug in the treatment of this subset of rare but highly aggressive gynecological tumors . P00747 kringle 5 reduces vascular leakage in the retina in rat models of oxygen-induced retinopathy and diabetes . AIMS/HYPOTHESIS : Retinal vascular leakage is an early pathological feature in diabetic retinopathy and can lead to macular oedema and loss of vision . Previously we have shown that plasminogen kringle 5 ( P13647 ) , an angiogenic inhibitor , inhibits retinal neovascularisation in the rat model of oxygen-induced retinopathy ( OIR ) . The purpose of this study was to examine the effect of P13647 on vascular leakage in the retina . METHODS : Neonatal rats were exposed to hyperoxia to induce OIR . Diabetes was induced in adult rats by injecting streptozotocin . Vascular permeability was measured by Evans blue method . Expression of vascular endothelial growth factor ( P15692 ) was evaluated using immunohistochemistry and western blot analysis . RESULTS : Rats with OIR and diabetes showed abnormal vascular hyperpermeability in the retina and iris . Intravitreal injection of P13647 , reduced vascular permeability in both animal models , but did not affect permeability in normal rats . P13647 reduced vascular permeability at doses substantially lower than that required for inhibition of retinal neovascularisation . The P13647 -induced reduction in vascular permeability correlated with its down-regulation of P15692 expression in the retina . Moreover , P13647 inhibited DB01277 -induced hyperpermeability , which is known to arise through up-regulation of endogenous P15692 expression . However , P13647 had no effect on the hyperpermeability induced by injection of exogenous P15692 . CONCLUSIONS/INTERPRETATION : Very low doses of P13647 reduce pathological vascular leakage in the retina . P13647 thus has therapeutic potential in the treatment of diabetic macular oedema . This effect can be ascribed , at least in part , to the down-regulation of endogenous P15692 expression . P15692 -dependent and PDGF-dependent dynamic neurovascular reconstruction in the neurohypophysis of adult mice . Hypothalamo-neurohypophysial system ( HNS ) releases arginine vasopressin ( AVP ) and oxytocin ( P01178 ) from axonal terminals of the neurohypophysis ( NH ) into blood circulation for controlling body fluid homeostasis and lactation . Chronic osmotic and suckling stimulations have been shown to cause neurovascular and neuroglial reconstruction in the NH of adult mammals and no study has been reported for vascular dynamics . The aim of this study was to elucidate the occurrence of continuous angiogenesis and growth factor-dependent neurovascular reconstruction in the NH of adult mice . Active proliferation of endothelial cells and oligodendrocyte progenitor cells ( OPCs ) was observed using the immunohistochemistry of bromodeoxyuridine and Ki-67 . P15692 ( P15692 ) and P15692 receptor 2 ( P35968 ( P35968 ) ) were highly expressed at pituicytes and endothelial cells respectively . Moreover , prominent expression of platelet-derived growth factor B ( PDGFB ) and PDGF receptor beta was observed at P01178 -containing axonal terminals and pericytes respectively . Administration of the selective tyrosine kinase inhibitor DB04849 for VEGFRs and STI571 for PDGFRs significantly decreased proliferation of endothelial cells and OPCs . Moreover , DB04849 treatment decreased vascular density by facilitating apoptosis of endothelial cells and the withdrawal of its treatment led to remarkable rebound proliferation of endothelial cells , so that vascular density rapidly returned to normal levels . DB04849 decreased the density of both AVP- and P01178 -containing axonal terminals , whereas STI571 selectively decreased the density of AVP-containing ones . Thus , this study demonstrates that the signaling pathways of P15692 and PDGF are crucial mediators for determining proliferation of endothelial cells and OPCs and the density of AVP- and P01178 -containing axonal terminals in the HNS . Antibodies to the epidermal growth factor receptor in non small cell lung cancer : current status of matuzumab and panitumumab . DB05101 and panitumumab are antibodies against the epidermal growth factor receptor ( P00533 ) that are being evaluated in several malignancies including non-small cell lung cancer ( NSCLC ) . In phase I trials of single-agent matuzumab in patients with P00533 -positive cancer , three tumor responses were documented in esophageal squamous cell carcinoma as well as colorectal carcinoma . A phase I trial of matuzumab in combination with paclitaxel has been reported in 18 patients with P00533 -positive advanced NSCLC . Objective responses were seen in 4 of 18 ( 23 % ) patients . A randomized phase II trial is currently ongoing in second-line NSCLC with matuzumab in combination with pemetrexed . A large dose/schedule trial of single-agent panitumumab enrolled 96 patients with P00533 -positive solid tumors . No responses were seen in the 14 lung cancer patients evaluated ; 5 of 39 patients with colorectal cancers had objective responses . A randomized phase II trial of carboplatin/paclitaxel with or without panitumumab in 166 patients with previously untreated advanced stage IIIB/IV NSCLC did not find any benefit for the panitumumab arm compared with the chemotherapy alone arm with regard to response rates , time to disease progression , or median survival time . The lack of a biomarker to identify a subset of NSCLC patients who may derive benefit from this agent limits any potential enthusiasm for further trials of panitumumab at this time in NSCLC . Effects of LSD on Ca++ currents in central 5-HT-containing neurons : P08908 receptors may play a role in hallucinogenesis . Drugs that influence the activity of central serotonergic neurons by activating a 5-hydroxytryptamine subtype of receptor ( P08908 ) alter mood and perception . Previously , we demonstrated with whole-cell recordings from acutely isolated 5-HT-containing dorsal raphe ( DR ) neurons from the adult rat that 5-HT inhibited Ca++ current and activated K+ current in DR neurons . We now show that DB04829 ( LSD ) mimics the actions of 5-HT ; it dramatically suppresses Ca++ current in a dose-dependent manner and activates an inwardly rectifying K+ conductance . Spiperone ( 0.2 microM ) , a P08908 /5-HT2 antagonist , blocks the effect of both LSD and 5-HT . The nonhallucinogenic structural analog 2-bromo-LSD ( 2-Bol ) at 10 microM has no effect on either Ca++ or K+ current by itself , but it competitively antagonizes both effects of LSD . Inhibition of 5-HT release resulting from P08908 receptor activation may play an integral role in the hallucinogenic actions of LSD by reducing competition between 5-HT and LSD for the postsynaptic 5-HT receptors . DB06779 , a low-molecular-weight heparin , promotes angiogenesis mediated by heparin-binding P15692 in vivo . Tumors are angiogenesis dependent and vascular endothelial growth factor-A ( P15692 ) , a heparin-binding protein , is a key angiogenic factor . As chemotherapy and co-treatment with anticoagulant low-molecular-weight heparin ( LMWH ) are common in cancer patients , we investigated whether angiogenesis in vivo mediated by P15692 is modulated by metronomic-type treatment with : ( i ) the LMWH dalteparin ; ( ii ) low-dosage cytostatic epirubicin ; or ( iii ) a combination of these two drugs . Using the quantitative rat mesentery angiogenesis assay , in which angiogenesis was induced by intraperitoneal injection of very low doses of P15692 , dalteparin sodium ( Fragmin(®) ) and epirubicin ( Farmorubicin(®) ) were administered separately or in combination by continuous subcutaneous infusion at a constant rate for 14 consecutive days . DB06779 was administered at 27 , 80 , or 240 IU/kg/day , i.e. , doses that reflect the clinical usage of this drug , while epirubicin was given at the well-tolerated dosage of 0.4 mg/kg/day . While dalteparin significantly stimulated angiogenesis in an inversely dose-dependent manner , epirubicin did not significantly affect angiogenesis . However , concurrent treatment with dalteparin and epirubicin significantly inhibited angiogenesis . The effect of dalteparin is the first demonstration of a proangiogenic effect of any LMWH in vivo . The fact that co-treatment with dalteparin and epirubicin significantly inhibited angiogenesis suggests a complex drug effect . 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 .

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interacts_with DB06684?

multiple_choice

P08069 targeted therapeutics : novel compounds and novel treatment strategies for cancer medicine . The insulin-like growth factor 1 receptor ( IGF-1R ) and its associated signalling system has provoked considerable interest over recent years as a novel therapeutic target in cancer . A brief outline of the IGF-1R signalling system and the rationale for its use in cancer medicine is given . This is followed by a discussion of the different possible targets within the IGF-1R system , and drugs developed to interact at each target . A systems-based approach is then used to review the in vitro and in vivo data in the published literature of the following compounds targeting IGF-1R components using specific examples : growth hormone releasing hormone antagonists ( e.g. JV-1-38 ) , growth hormone receptor antagonists ( e.g. pegvisomant ) , IGF-1R antibodies ( e.g. CP-751,871 , AVE1642/EM164 , DB05759 , P35240 -717454 , BIIB022 , Q99217 479 , MK-0646/h7C10 ) , and IGF-1R tyrosine kinase inhibitors ( e.g. BMS-536942 , BMS-554417 , DB00238 -AEW541 , DB00238 -ADW742 , AG1024 , potent quinolinyl-derived imidazo (1,5-a)pyrazine PQIP , picropodophyllin PPP , Nordihydroguaiaretic acid Insm-18/NDGA ) . The following tumour types are specifically discussed : lung , breast , colorectal , pancreatic , NETs , sarcoma , prostate , leukaemia , multiple myeloma . Other tumour types are mentioned briefly : squamous cell carcinoma of the head and neck , melanoma , glioblastoma , ovary , gastric and mesothelioma . Results of early stage clinical trials , involving recently patented drugs. are included where appropriate . We then outline the current understanding of toxicity related to IGF-1R targeted therapy , and finally outline areas for further research . DB02709 protects against peripheral deficits in a mouse model of Huntington 's disease . Sirtuins are NAD-dependent deacetylases that regulate important biologic processes including transcription , cell survival and metabolism . Activation of Q96EB6 , a mammalian sirtuin , extends longevity and increases neuronal survival . An important substrate of Q96EB6 is peroxisome proliferator-activated receptor gamma co-activator-1alpha ( P20142 -1alpha ) , a principal regulator of energy metabolism , whose function is significantly impaired in Huntington 's disease ( HD ) . We studied the effects of a pharmacological preparation of the Q96EB6 activator resveratrol ( DB05073 -M ) , in the N171-82Q transgenic mouse model of HD . We analyzed motor performance , survival , central and peripheral pathology and levels of P20142 -1alpha expression . Administration of DB05073 -M increased expression of P20142 -1alpha , as well as its downstream targets , nuclear respiratory factor-1 ( Q16656 ) and uncoupling protein-1 ( P25874 -1 ) in brown adipose tissue ( Q14032 ) , but there was no effect on P20142 -1alpha , Q16656 or the mitochondrial transcription factor ( Tfam ) in the striatum . DB05073 -M administration also reduced Q14032 vacuolation and decreased elevated blood glucose levels . However , there was no significant improvement in weight loss , motor performance , survival and striatal atrophy . Activation of the P20142 -1alpha signaling pathway via resveratrol-induced activation of Q96EB6 , therefore , is an effective therapy in Q14032 , but not in the central nervous system of HD transgenic mice . Proteolytic cleavage of platelet endothelial cell adhesion molecule-1 ( P16284 /CD31 ) is regulated by a calmodulin-binding motif . Homophilic engagement of platelet endothelial cell adhesion molecule-1 ( P16284 /CD31 ) induces ' outside-in ' signal transduction that results in phosphorylation events and recruitment and activation of signalling molecules . The formation of signalling scaffolds with P16284 are important signalling events that modulate platelet secretion , aggregation and platelet thrombus formation . In this study , we describe a novel interaction between P16284 and cytosolic calmodulin ( P62158 ) in platelets . Reciprocal co-immunoprecipitation studies revealed that cytosolic P62158 is constitutively associated with P16284 in resting , thrombin activated and aggregated human platelets . Our studies demonstrate that P62158 directly interacts with a P16284 peptide ( 594-604 ) C595A containing the sequences (594)KAFYLRKAKAK(604) . This P62158 : P16284 interaction has a threefold higher affinity than P62158 : Q9HCN6 interaction . It is potentiated by the addition of calcium ions , and dissociated by the P62158 inhibitor , trifluoperazine . Treatment of platelets with P62158 inhibitors triggers cleavage of P16284 in a time- and dose-dependent manner . Furthermore , this membrane proximal portion of P16284 is conserved across mammalian species and the helical representation of basic/hydrophobic residues reveals a charge distribution analogous to other P62158 -binding motifs in other proteins . Taken together , these results suggest that this highly charged cluster of amino acids in the P16284 cytoplasmic domain directly interacts with P62158 and this novel interaction appears to regulate cleavage of P16284 . P23560 activation of P62158 -kinase kinase via transient receptor potential canonical channels induces the translation and synaptic incorporation of P42261 -containing calcium-permeable AMPA receptors . Glutamatergic synapses in early postnatal development transiently express calcium-permeable AMPA receptors ( CP-AMPARs ) . Although these P42262 -lacking receptors are essential and are elevated in response to brain-derived neurotrophic factor ( P23560 ) , little is known regarding molecular mechanisms that govern their expression and synaptic insertion . Here we show that P23560 -induced P42261 translation in rat primary hippocampal neurons requires the activation of mammalian target of rapamycin ( P42345 ) via calcium calmodulin-dependent protein kinase kinase ( CaMKK ) . Specifically , P23560 -mediated phosphorylation of threonine 308 ( T308 ) in AKT , a known substrate of CaMKK and an upstream activator of P42345 -dependent translation , was prevented by ( 1 ) pharmacological inhibition of CaMKK with STO-609 , ( 2 ) overexpression of a dominant-negative CaMKK , or ( 3 ) short hairpin-mediated knockdown of CaMKK . P42261 surface expression induced by P23560 , as assessed by immunocytochemistry using an extracellular N-terminal P42261 antibody or by surface biotinylation , was impaired following knockdown of CaMKK or treatment with STO-609 . Activation of CaMKK by P23560 requires transient receptor potential canonical ( TRPC ) channels as SKF-96365 , but not the DB01221 receptor antagonist d-APV , prevented P23560 -induced P42261 surface expression as well as phosphorylation of CaMKI , AKT(T308) , and P42345 . Using siRNA we confirmed the involvement of Q9UL62 and Q9Y210 subunits in P23560 -induced AKT(T308) phosphorylation . The P23560 -induced increase in mEPSC was blocked by IEM-1460 , a selected antagonist of CP-AMPARs , as well as by the specific repression of acute P42261 translation via siRNA to P42261 but not P42262 . Together these data support the conclusion that newly synthesized P42261 subunits , induced by P23560 , are readily incorporated into synapses where they enhance the expression of CP-AMPARs and synaptic strength . Genetic polymorphism and activities of human lung alcohol and aldehyde dehydrogenases : implications for ethanol metabolism and cytotoxicity . DB00898 dehydrogenase ( DB00067 ) and aldehyde dehydrogenase ( ALDH ) exhibit genetic polymorphism and tissue specificity . DB00067 and ALDH isozyme phenotypes from 39 surgical Chinese lung specimens were identified by agarose isoelectric focusing . The identity of the lung beta-ADHs was further demonstrated by their characteristic pH-activity profiles for ethanol oxidation , Km values for NAD and ethanol , and inhibition by DB01213 or 1,10-phenanthroline . The beta 2 allele , coding for beta 2 polypeptide , was found to be predominant in the lung specimens studied . The DB00067 activities in the lungs with the homozygous phenotype P00325 2-2 ( exhibiting beta 2 beta 2 ) and P00325 1-1 ( exhibiting beta 1 beta 1 ) and the heterozygous phenotype P00325 P35326 ( exhibiting beta 2 beta 2 , beta 2 beta 1 , and beta 1 beta 1 ) were determined to be 999 +/- 77 , 48 +/- 17 , and 494 +/- 61 nmol/min/g tissue , respectively . Fifty-one percent of the specimens studied lacked the P05091 activity band on the isoelectric focusing gels . The activities in the lung tissues with the P05091 -active phenotype and the inactive phenotype were determined to be 30 +/- 3 and 17 +/- 1 nmol/min/g tissue , respectively . These findings indicate that human pulmonary ethanol-metabolizing activities differ significantly with respect to genetic polymorphism at both the P00325 and the P05091 loci . The results suggest that individuals with high Vmax beta 2- DB00067 and deficient in low-Km mitochondrial P05091 , accounting for approximately 45 % of the Chinese population , may end up with acetaldehyde accumulation during alcohol consumption , rendering them vulnerable to tissue injury caused by this highly reactive and toxic metabolite . Molecular identification of the human O75899 : cell surface expression and coupling to adenylyl cyclase in the absence of Q9UBS5 . We have identified a gene encoding a GABAB receptor , the human O75899 , located on chromosome 9q22.1 , that is distinct from the recently reported rat Q9UBS5 . O75899 structurally resembles Q9UBS5 ( 35 % identity ) , having seven transmembrane domains and a large extracellular region , but differs in having a longer carboxy-terminal tail . O75899 is localized to the cell surface in transfected COS cells , and negatively couples to adenylyl cyclase in response to GABA , baclofen , and 3-aminopropyl(methyl)phosphinic acid in CHO cells lacking Q9UBS5 . DB00181 action is inhibited by the GABABR antagonist , 2-hydroxysaclofen . The human O75899 and Q9UBS5 genes are differentially expressed in the nervous system , with the greatest difference being detected in the striatum in which Q9UBS5 but not O75899 mRNA transcripts are detected . O75899 and Q9UBS5 mRNAs are also coexpressed in various brain regions such as the Purkinje cell layer of the cerebellum . Identification of a functional homomeric O75899 coupled to adenylyl cyclase suggests that the complexity of GABAB pharmacological data is at least in part due to the presence of more than one receptor and opens avenues for future research leading to an understanding of metabotropic GABA receptor signal transduction mechanisms . Adenovirus-mediated Q16763 P25874 gene transfer prevents autoamputation in a mouse model of hindlimb ischemia . Q16763 ubiquitin carrier protein ( P25874 ) stabilizes hypoxia-inducible factor-1α ( HIF-1α ) inducing ischemic vascular responses . Here , we investigated the effect of P25874 gene transfer on therapeutic angiogenesis . Adenovirus-encoded P25874 ( Ad-F- P25874 ) increased the expression of vascular endothelial growth factor ( P15692 ) and fibroblast growth factor-2 ( P09038 ) in cells and mice . Conditioned media from P25874 -overexpressing cells promoted proliferation , tubule formation , and invasion of human umbilical-vascular-endothelial cells ( HUVECs ) , and vascularization in chorioallantoic membrane ( P62158 ) assay . Ad-F- P25874 increased the vessel density in the Martigel plug assay , and generated copious vessel-like structures in the explanted muscle . The P25874 effect on angiogenesis was dependent on P15692 and P09038 . In mouse hindlimb ischemia model ( N = 30/group ) , autoamputation ( limb loss ) occurred in 87 % and 68 % of the mice with saline and Ad encoding β-galactosidase ( Ad-LacZ ) , respectively , whereas only 23 % of the mice injected with Ad-F- P25874 showed autoamputation after 21 days of treatment . Ad-F- P25874 increased protein levels of HIF-1α , platelet-endothelial cell adhesion molecule-1 ( P16284 ) , smooth muscle cell actin ( SMA ) in the ischemic muscle , and augmented blood vessels doubly positive for P16284 and SMA . Consequently , P25874 gene transfer prevented muscle degeneration and autoamputation of ischemic limb . The results suggest that Q16763 P25874 may be a target for therapeutic angiogenesis . Evidence that 5-HT2c receptor antagonists are anxiolytic in the rat Geller-Seifter model of anxiety . Four non-selective P28335 /5- Q13049 receptor antagonists , mianserin ( 2-8 mg/kg ) , 1-naphthyl piperazine ( 1-NP ) ( 0.5-1 mg/kg ) , ICI 169,369 ( 20 mg/kg ) and LY 53857 ( 5 mg/kg ) , increased punished responding for a food reward in the rat Geller-Seifter test 30 min after subcutaneous ( SC ) administration . This property was shared by the benzodiazepine anxiolytic chlordiazepoxide ( 5 mg/kg SC ) . However , the selective 5- Q13049 receptor antagonists ketanserin ( 0. P35326 mg/kg SC ) and altanserin ( 0.5 , 1 mg/kg SC ) had little effect . The P08908 , P28222 and beta-adrenergic receptor antagonists pindolol and cyanopindolol ( 6 mg/kg SC ) did not affect punished responding either , nor did the P28221 receptor partial agonist and alpha 2 adrenergic receptor antagonist yohimbine ( 2.5 mg/kg SC ) or the histamine H1 receptor antagonist mepyramine ( 1 mg/kg SC ) . Unpunished responding was also modestly increased after some doses of the P28335 /5- Q13049 receptor antagonists . However , this effect was inconsistent and was also seen after chlordiazepoxide . Furthermore , it was not associated with the increase in punished responding observed in rats orally treated with mianserin ( 10 , 20 mg/kg ) , 1-NP ( 10 , 20 mg/kg ) or ICI 169,369 ( 50 mg/kg ) . The action of the P28335 /5- Q13049 receptor antagonists tested is therefore consistent with anxiolysis . The results also strongly suggest that this effect is mediated by blockade of the P28335 receptor , although the possibility of P41595 receptor mediation is discussed . Effects of systemic injections of vilazodone , a selective serotonin reuptake inhibitor and serotonin 1A receptor agonist , on anxiety induced by predator stress in rats . We examined the effect of DB06684 , a selective serotonin reuptake inhibitor ( SSRI ) and serotonin 1A ( 5-HT(1A) ) receptor agonist [ Bartoszyk , G.D. , Hegenbart , R. , Ziegler , H. , 1997. P50402 68843 , a serotonin reuptake inhibitor with selective presynaptic P08908 receptor agonistic properties. Eur. J. Pharmacol. 322 , 147-153. ] , on change in affect following predator stress . DB06684 and vehicle injection ( intraperitoneal ) occurred either 10 min after predator stress ( prophylactic testing ) , or 90 min prior to behavioral testing for the effects of predator stress ( therapeutic testing ) . Predator stress involved unprotected exposure of rats to a domestic cat . Behavioral effects of stress were evaluated with hole board , plus-maze , and acoustic startle tests 1 week after stress . Predator stress increased anxiety-like behavior in the plus-maze and elevated response to acoustic startle . In prophylactic testing , DB06684 affected stress potentiation of startle at doses above 5 mg/kg . DB06684 increased stress elevation of startle at 10 mg/kg . Higher doses of DB06684 ( 20 and 40 mg/kg ) blocked stress potentiation of startle . In contrast , DB06684 had no effect on stress potentiation of anxiety in the plus-maze . In therapeutic testing , DB06684 increased stress elevation of startle at all doses . In contrast , therapeutic DB06684 had no effect on stress potentiation of anxiety in the plus-maze . Taken together , the data suggest a prophylactic potential for DB06684 in the treatment of changes in hypervigilance following severe stress . The contribution of serotonin P28335 and melanocortin-4 receptors to the satiety signaling of glucagon-like peptide 1 and liraglutide , a glucagon-like peptide 1 receptor agonist , in mice . Glucagon-like peptide 1 ( P0C6A0 ) , an insulinotropic gastrointestinal peptide produced mainly from intestinal endocrine L-cells , and liraglutide , a P43220 ( P43220 ) agonist , induce satiety . The serotonin P28335 receptor ( 5-HT2CR ) and melanoroctin-4 receptor ( P32245 ) are involved in the regulation of food intake . Here we show that systemic administration of P0C6A0 ( 50 and 200μg/kg ) -induced anorexia was blunted in mice with a 5HT2CR null mutation , and was attenuated in mice with a heterozygous P32245 mutation . On the other hand , systemic administration of liraglutide ( 50 and 100μg/kg ) suppressed food intake in mice lacking 5-HT2CR , mice with a heterozygous mutation of P32245 and wild-type mice matched for age . Moreover , once-daily consecutive intraperitoneal administration of liraglutide ( 100μg/kg ) over 3days significantly suppressed daily food intake and body weight in mice with a heterozygous mutation of P32245 as well as wild-type mice . These findings suggest that P0C6A0 and liraglutide induce anorexia via different central pathways . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Inhibition of noradrenaline release via presynaptic P28222 receptors of the rat vena cava . In the rat inferior vena cava preincubated with 3H-noradrenaline , the effects of nine serotonin ( 5-HT ) receptor agonists and of eight antagonists ( including two beta-adrenoceptor blocking agents ) on the electrically evoked 3H overflow were determined . 1 . 5-HT , 5-carboxamido-tryptamine , 5-methoxy-3(1,2,3,6-tetrahydropyridine-4-yl)-1H-indole ( RU 24969 ) , 5-methoxytryptamine , N,N-dimethyl-5HT , tryptamine and 5-aminotryptamine inhibited the evoked 3H overflow . The potencies of these agonists in inhibiting overflow were significantly correlated with their affinities for P28222 binding sites , but not with their affinities for P08908 , P28335 or 5-HT2 binding sites . 8-Hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) , a P08908 receptor agonist , and ipsapirone , a partial agonist at these receptors , did not inhibit overflow . 2 . Cyanopindolol facilitated the evoked 3H overflow , an effect which was abolished by propranolol . The maximum inhibition of overflow obtainable with 5-HT was diminished by cyanopindolol . 3 . The concentration-response curve for 5-HT was shifted to the right by metitepine , metergoline , quipazine , 6-chloro-2-(1-piperazinyl)pyrazine ( MK 212 ) and propranolol which , given alone , did not affect 3H overflow . The apparent pA2 values of these antagonists tended to be correlated with their affinities for P28222 ( but not P08908 , P28335 or 5-HT2 ) binding sites . Ketanserin , a 5-HT2 receptor antagonist , and spiperone , which blocks 5-HT2 and P08908 but not P28222 or P28335 receptors , failed to antagonize the effect of 5-HT. ( ABSTRACT TRUNCATED AT 250 WORDS ) DB05332 administration shows reduced megakaryocyte response-capacity and increased myelofibrosis in a mouse model of P35579 -RD . Macrothrombocytopenia in P35579 -related disease ( P35579 -RD ) results from defects in nonmuscular myosin-IIA function . P40238 agonists ( eltrombopag ; romiplostim ) seem to improve hemostasis , but little is known about their biologic effects in P35579 -RD . We administered romiplostim to Myh9(-/-) mice ( 100 μg/kg , every 3 days , during 1 month ) . MKs increased to similar numbers in Myh9(-/-) and wild-type ( WT ) mice ( with an increase in immature MKs ) , but Myh9(-/-) platelet count response was much less ( 2.5-fold vs 8-fold increase ) . A strong increase in MK nuclei emboli in the lung , in WT and Myh9(-/-) mice , indicates increased transmigration of MKs from the BM . Prolonged ( but not acute ) treatment with romiplostim decreased expression of GPIb-IX-V complex and Q9HCN6 , but not of GPIIbIIIa , and bleeding time increased in WT mice . Microcirculation was not altered by the increased number of large platelets in any of the assessed organs , but in Myh9(-/-) mice a much stronger increase in BM reticulin fibers was present after 4 weeks of romiplostim treatment vs WT mice . These data further encourage short-term use of thrombopoietic agents in patients with P35579 -RDs ; however , myelofibrosis has to be considered as a potential severe adverse effect during longer treatment . Reduction of GPIbIX/ Q9HCN6 expression by romiplostim requires further studies . P43220 agonists and the thyroid : C-cell effects in mice are mediated via the P43220 and not associated with P07949 activation . DB06655 and exenatide are glucagon-like peptide receptor ( P43220 ) agonists used in the treatment of type 2 diabetes . Both molecules have been associated with the development of thyroid C-cell tumors after lifetime exposure in rodents . Previously , it has been reported that these tumors are preceded by increased plasma calcitonin and C-cell hyperplasia . We can now document that the murine C-cell effects are mediated via P43220 . Thus , 13 wk of continuous exposure to P43220 agonists was associated with marked increases in plasma calcitonin and in the incidence of C-cell hyperplasia in wild-type mice . In contrast , similar effects were not seen in P43220 knockout mice . Human C-cell cancer is often caused by activating mutations in the rearranged-during-transfection ( P07949 ) protooncogene . We developed an immunohistochemical method to assess P07949 activation in tissues . DB06655 dosing to mice was not found to activate P07949 . Further evaluation of the signaling pathways demonstrated that liraglutide increased ribosomal S6 , but not MAPK kinase , phosphorylation . These observations are consistent with effects of P43220 agonists on rodent C cells being mediated via mammalian target of rapamycin activation in a P07949 - and MAPK-independent manner . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . Role of presynaptic serotonergic receptors on the mechanism of action of P08908 and P28222 agonists on masculine sexual behaviour : physiological and pharmacological implications . In order to establish whether the P08908 or the 5HT1B agonists , 8-OH-DPAT or TFMPP , produce their facilitatory or inhibitory actions on masculine sexual behaviour via a mechanism involving : ( a ) the serotonin synthesis or release ; ( b ) the stimulation of presynaptic receptors , or ( c ) the stimulation of somatodendritic receptors , three series of experiments were performed . The administration of the serotonin synthesis inhibitor , p-chlorophenylalanine ( p- P15085 , 300 mg/kg x 3 days ) , facilitated sexual behaviour but does not interfere neither with the inhibitory nor with the facilitatory effects of TFMPP ( 0.5 mg/kg ) or 8-OH-DPAT ( 0.5 mg/kg ) , respectively . The icv or the intraraphé administration of the serotonergic neurotoxin , 5,7-dihydroxytryptamine ( 5,7- DB02901 ) , slightly stimulated masculine sexual behaviour and produced a decrease in serotonin and its metabolite levels . In lesioned animals TFMPP ( 0.5 mg/kg ) resulted in an inhibitory effect reflected as a prolongation of the ejaculation latency . The inhibitory effect of this drug on mounting behaviour was not observed in 5,7- DB02901 treated rats . In lesioned animals 8-OH-DPAT ( 0.5 mg/kg ) produced the same facilitatory effect . Present data indicate that serotonergic postsynaptic receptors mediate both the inhibitory and the facilitatory actions of TFMPP or 8-OH-DPAT in copulation . All data further support the idea that endogenous serotonin acts via the stimulation of P28222 receptors to induce its inhibitory effects on masculine sexual behaviour . Light and X-ray scattering show decorin to be a dimer in solution . P07585 is a widely distributed member of the extracellular matrix small leucine-rich repeat glycoprotein/proteoglycan family . For investigation of its physical properties , decorin from two sources ( young steer skin and a recombinant adenovirus ) was used . The first sample was extracted into 7 m urea and purified , while the second was isolated from medium conditioned by 293A cells infected with adenovirus and purified without chaotropes . The only chemical differences detected between these materials were a slightly shorter glycosaminoglycan chain and the retention of the propeptide on the latter . Circular dichroism spectra of the two samples were virtually identical , showing a high proportion of beta-sheet and beta-turn and little alpha-helix . The protein cores were completely denatured in 2.25 m guanidine HCl ( GdnHCl ) but recovered their secondary structure on removal of chaotrope . Light scattering of material eluted from gel-filtration columns in DB03754 -buffered saline , pH 7.0 , gave molecular mass values of 165 +/- 1 kDa and 84.6 +/- 4 kDa for intact decorin and the glycoprotein core produced by digestion with chondroitin ABC lyase , respectively . Intact recombinant prodecorin had a mass of 148 +/- 18 kDa . These values , which are double those estimated from SDS gel electrophoresis or from the known sequences and compositions , were halved in 2.5 m GdnHCl . Data from solution x-ray scattering of intact decorin and its core in DB03754 -buffered saline are consistent with a dimeric particle whose protein component has a radius of gyration of 31.6 +/- 0.4 A , a maximum diameter of 98 +/- 5 A , and approximates two intertwined C shapes . Computation of standard binding free energies of polar and charged ligands to the glutamate receptor P42262 . Accurate calculation of the binding affinity of small molecules to proteins has the potential to become an important tool in rational drug design . In this study , we use the free energy perturbation ( FEP ) method with restraints to calculate the standard binding free energy of five ligands ( ACPA , AMPA , CNQX , DB03759 , and glutamate ) to the glutamate receptor P42262 , which plays an essential role in synaptic transmission . To deal with the convergence problem in FEP calculations with charged ligands , we use a protocol where the ligand is coupled in the binding site while it is decoupled in bulk solution simultaneously . The contributions from the conformational , rotational , and translational entropies to the standard binding free energy are determined by applying/releasing respective restraints to the ligand in bulk/binding site . We also employ the confine-and-release approach , which helps to resolve convergence problems in FEP calculations . Our results are in good agreement with the experimental values for all five ligands , including the charged ones which are often problematic in FEP calculations . We also analyze the different contributions to the binding free energy of each ligand to P42262 and discuss the nature of these interactions . P25116 14-amino acid peptide mediates endothelial hyperadhesivity and neutrophil adhesion by P16109 -dependent mechanism . Thrombin cleaves its receptor at arginine-41 , resulting in the generation of a new receptor NH2-terminus with the sequence SFLLRNPNDKYEPF . This peptide ( TRP-14 ) may signal a variety of thrombin 's responses . We examined the effects of TRP-14 in inducing endothelial cell hyperadhesivity and neutrophil ( PMN ) adhesion to endothelial cell monolayers . Human umbilical vein endothelial cells ( HUVECs ) challenged with TRP-14 ( 10(-4) to 10(-5) M ) produced concentration-dependent increases in endothelial adhesivity to PMN . In contrast , position 1 to 2 inverted peptide ( FSLLRNPNDKYEPF ) did not induce the response . The adhesion response was transient ; that is , PMN adhesion increased within 15 minutes and decreased by 75 minutes after TRP-14 challenge of HUVECs . The transient endothelial adhesiveness paralleled the time course of P16109 expression . TRP-14-induced release of P16109 from intracellular stores may be a critical determinant of the response since treatment of endothelial cells with anti- P16109 monoclonal antibody ( mAb ) P55008 prevented the increase in PMN adhesion . Control nonneutralizing anti- P16109 mAb P28222 and mAb P23921 /1 directed against intercellular adhesion molecule-1 ( P05362 ) on HUVECs were ineffective . The results indicate that the " tethered ligand " of the thrombin receptor created by the proteolytic action of thrombin on its receptor ( i.e. , TRP-14 ) signals increased endothelial adhesiveness by a P16109 -dependent mechanism . Thrombin-induced PMN adhesion may involve formation of a new NH2-terminus of the endothelial thrombin receptor with the sequence SFLLRNPNDKYEPF followed by activation of endothelial second messenger pathways and the transient expression of P16109 . Screening for candidate gene regions in narcolepsy using a microsatellite based approach and pooled DNA . Narcolepsy is a complex sleep disorder characterized by excessive daytime sleepiness and cataplexy . Mutations in genes of the hypocretin ( orexin ) neurotransmitter system cause narcoleptic symptoms in animal models . The absence of hypocretin in the cerebrospinal fluid of human patients is hypothesized to originate from destruction of hypocretinergic cells in the hypothalamus , the cause of which remains unknown . Due to strong HLA association autoimmune models of narcolepsy pathogenesis are still mostly favored . Genetic predisposition factors other than HLA are likely to play a role in causing the disorder . We screened three sets of gene regions ( n=254 ) for association with narcolepsy using a microsatellite based approach and pooled DNA : genes related to immunity , particularly apoptosis ; genes related to regulation of circadian rhythmicity ; genes coding for several factors of neurotransmission . In relation to apoptosis an association was found for the Q99933 gene region . Interestingly , microsatellites representing four genomic regions related to neurotransmission revealed association with narcolepsy : P21964 , P14416 , Q9UBS5 , and P28223 . These results , although exploratory and still to be confirmed in independent samples , support a complex pathogenetic model for narcolepsy , including disturbances of neurotransmission rather than involvement of autoimmunity . Modulatory effects of serotonin on glutamatergic synaptic transmission and long-term depression in the deep cerebellar nuclei . The deep cerebellar nuclei ( P07585 ) are the terminal components of the cerebellar circuitry and constitute its primary output structure . Their activity is important for certain forms of motor learning as well as generation and control of movement . P07585 neurons receive glutamatergic excitatory inputs from the pontine nuclei via mossy fibres ( MFs ) and concomitantly receive inputs from 5-HT-containing neurons of the raphe nuclei . We aimed to explore the roles of 5-HT at MF- P07585 synapses by using cerebellar slices from 11 to 15-day-old rats . Bath application of 5-HT reversibly decreased the amplitude of stimulation-evoked excitatory postsynaptic currents ( eEPSCs ) via the activation of P28222 receptors at the presynaptic terminals of the MFs . Burst stimulation of the MFs elicited long-term depression ( LTD ) at the MF- P07585 synapses that require activation of the group I metabotropic glutamate receptor ( mGluR ) . In the presence of 5-HT , the extent of burst-induced LTD of MF EPSCs was significantly reduced . Application of 5-HT also decreased the amplitude of mGluR-dependent slow EPSCs evoked by similar burst stimulation . Furthermore , ( S ) -3,5-dihydroxyphenylglycine ( DHPG ) , a group I mGluR agonist , induced chemical LTD of MF EPSCs , and 5-HT had no significant effect on this LTD . Taken together , the results suggest that 5-HT not only has transitory inhibitory effects on MF EPSCs but also plays a role in regulating the long-term synaptic efficacy . Thrombin receptors and their antagonists : an update on the patent literature . IMPORTANCE OF THE FIELD : Thrombin plays a central role in cardiovascular inflammation . Most of the cellular responses to thrombin are mediated by cell surface protease-activated receptors ( PARs ) . Several preclinical studies indicate that PARs are potential targets for treating cardiovascular diseases such as thrombosis , atherosclerosis and restenosis . Among PARs , P25116 has emerged as an important therapeutic target . AREAS COVERED IN THIS REVIEW : This review covers recent advances in the development of thrombin receptors antagonists . It is focused on the search for P25116 antagonists as this is at the moment the most promising and attractive target . However , some early promising studies on PAR-3 and -4 antagonists are also reported . WHAT THE READER WILL GAIN : The review has been written in order to give to the reader hints and references that cover , in our opinion , the most interesting and/or promising approaches in this research field . TAKE HOME MESSAGE : Research on P25116 antagonists has finally led to good clinical candidates such as DB05692 ( Schering-Plough ) and E-5555 ( Eisai Co. ) . Clinical trials clearly demonstrate that development of PAR1 antagonists is not only possible but most likely will lead to development of antiplatelet drugs as well as of drugs useful for the treatment of inflammatory , proliferative and neurodegenerative diseases . Gene expression of malignant rhabdoid tumor cell lines by reverse transcriptase-polymerase chain reaction . Malignant rhabdoid tumors ( MRT ) are characterized by unique neoplastic cells demonstrating phenotypic diversity . By using the reverse transcriptase-polymerase chain reaction , we have detected expression of various genes before and after differentiation induction with four different agents in four established MRT cell lines ( TM87-16 , STM91-01 , TTC642 , and TTC549 ) . The agents used in this study were all-trans retinoic acid ( RA ) , 12-O-tetradecanoylphorbol-13-acetate ( TPA ) , interleukin-3 , or interferon-gamma . Before and after induction , c-myc , P01344 , P08069 , and P11717 were constitutively expressed by all four cell lines . The neurofilament medium-size ( P07197 ) was constitutively expressed by the TM87-16 and TTC642 , and the S100 protein alpha subunit was expressed by TM87-16 , TTC642 , and TTC549 . Chromogranin A was expressed by TM87-16 only after treatment with either TPA or RA . MyoD , N-myc , tyrosine hydroxylase , N- P62158 , trkA , and the S100 protein beta subunit were not expressed by any cell line before or after induction with these agents . All the MRT cell lines in this study except TM87-16 were highly resistant to differentiation induction . The proliferating cells in TM87-16 and TTC642 expressed mRNA profiles characteristic of neuroectoderm .

MH_train_1393

MH_train_1393

MH_train_1393

interacts_with DB08827?

multiple_choice

[ Protective effects of penehyclidine hydrochloride against acute renal injury induced by hemorrhagic shock and lipopolysaccharides in rats ] . OBJECTIVE : To investigate the effect of penehyclidine hydrochloride ( Q00325 ) in a rat model of renal injury induced by hemorrhagic shock and lipopolysaccharides ( LPS ) . METHODS : Forty-five healthy Wistar rats were randomized into sham operated group , model group , and 3 penehyclidine hydrochloride ( Q00325 ) dose ( 1 , 2 and 3 mg/kg ) groups ( P78364 , Q8IXK0 , and Q8NDX5 groups , respectively ) . The arterial blood samples were collected to determine the concentrations of serum tumor necrosis factor-α ( P01375 -α ) , interleukin-8 ( P10145 ) , interleukin-1 ( IL-1 ) , urine creatinine ( Cr ) and blood urine nitrogen ( BUN ) , and the renal tissues were collected to measure the expressions of P05362 and nuclear factor-κB ( NF-κB ) and observe the pathological changes . RESULTS : P01375 -α , P10145 , IL-1 , Cr , BUN , P05362 and NF-κB in the 3 Q00325 groups were significantly lower than those in the model group ( P < 0.05 ) . P01375 -α , P10145 , IL-1 , Cr and BUN were significantly lower in P78364 ( P < 0.05 ) than in the Q8IXK0 and Q8NDX5 groups , and P05362 and NF-κB were similar between 3 Q00325 groups ( P > 0.05 ) . Compared with the model group , the 3 Q00325 groups showed lessened pathological changes in the renal tubules . CONCLUSION : Q00325 has protective effects against renal injury induced by hemorrhagic-endotoxin shock in rats , and treatment with 1 mg/kg Q00325 produces the most significant protective effect . Niacin reduces plasma P11597 levels by diminishing liver macrophage content in P11597 transgenic mice . The anti-dyslipidemic drug niacin has recently been shown to reduce the hepatic expression and plasma levels of P11597 . Since liver macrophages contribute to hepatic P11597 expression , we investigated the role of macrophages in the P11597 -lowering effect of niacin in mice . In vitro studies showed that niacin does not directly attenuate P11597 expression in macrophages . Treatment of normolipidemic human P11597 transgenic mice , fed a Western-type diet with niacin for 4 weeks , significantly reduced the hepatic cholesterol concentration ( -20 % ) , hepatic P11597 gene expression ( -20 % ) , and plasma P11597 mass ( -30 % ) . Concomitantly , niacin decreased the hepatic expression of P34810 ( -44 % ) and P45844 ( -32 % ) , both of which are specific markers for the hepatic macrophage content . The decrease in hepatic P11597 expression was significantly correlated with the reduction of hepatic macrophage markers . Furthermore , niacin attenuated atherogenic diet-induced inflammation in liver , as evident from decreased expression of P01375 ( -43 % ) . Niacin similarly decreased the macrophage markers and absolute macrophage content in hyperlipidemic P02649 *3-Leiden. P11597 transgenic mice on a Western-type diet . In conclusion , niacin decreases hepatic P11597 expression and plasma P11597 mass by attenuating liver inflammation and macrophage content in response to its primary lipid-lowering effect , rather than by attenuating the macrophage P11597 expression level . Loss of both phospholipid and triglyceride transfer activities of microsomal triglyceride transfer protein in abetalipoproteinemia . Mutations in microsomal triglyceride transfer protein ( P55157 ) cause abetalipoproteinemia ( P00519 ) , characterized by the absence of plasma apoB-containing lipoproteins . In this study , we characterized the effects of various P55157 missense mutations found in P00519 patients with respect to their expression , subcellular location , and interaction with protein disulfide isomerase ( P07237 ) . In addition , we characterized functional properties by analyzing phospholipid and triglyceride transfer activities and studied their ability to support apoB secretion . All the mutants colocalized with calnexin and interacted with P07237 . We found that R540H and N780Y , known to be deficient in triglyceride transfer activity , also lacked phospholipid transfer activity . Novel mutants S590I and G746E did not transfer triglycerides and phospholipids and did not assist in apoB secretion . In contrast , D384A displayed both triglyceride and phospholipid transfer activities and supported apoB secretion . These studies point out that P00519 is associated with the absence of both triglyceride and phospholipid transfer activities in P55157 . The protein disulfide isomerase O95994 is essential for production of intestinal mucus . Protein disulfide isomerases ( PDIs ) aid protein folding and assembly by catalyzing formation and shuffling of cysteine disulfide bonds in the endoplasmic reticulum ( ER ) . Many members of the P07237 family are expressed in mammals , but the roles of specific PDIs in vivo are poorly understood . A recent homology-based search for additional P07237 family members identified anterior gradient homolog 2 ( O95994 ) , a protein originally presumed to be secreted by intestinal epithelial cells . Here , we show that O95994 is present within the ER of intestinal secretory epithelial cells and is essential for in vivo production of the intestinal mucin Q02817 , a large , cysteine-rich glycoprotein that forms the protective mucus gel lining the intestine . A cysteine residue within the O95994 thioredoxin-like domain forms mixed disulfide bonds with Q02817 , indicating a direct role for O95994 in mucin processing . Mice lacking O95994 were viable but were highly susceptible to colitis , indicating a critical role for O95994 in protection from disease . We conclude that O95994 is a unique member of the P07237 family , with a specialized and nonredundant role in intestinal mucus production . Mitigation of DB03128 Esterase Activity in the 1,7-Diazacarbazole Series of Inhibitors of Checkpoint Kinase 1 . Checkpoint kinase 1 ( ChK1 ) plays a key role in the DNA damage response , facilitating cell-cycle arrest to provide sufficient time for lesion repair . This leads to the hypothesis that inhibition of ChK1 might enhance the effectiveness of DNA-damaging therapies in the treatment of cancer . Lead compound 1 ( Q9Y223 -783 ) , the prototype of the 1,7-diazacarbazole class of ChK1 inhibitors , was found to be a highly potent inhibitor of acetylcholine esterase ( P22303 ) and unsuitable for development . A campaign of analogue synthesis established SAR delineating ChK1 and P22303 activities and allowing identification of new leads with improved profiles . In silico docking using a model of P22303 permitted rationalization of the observed SAR . Compounds 19 ( Q9Y223 -900 ) and 30 ( Q9Y223 -145 ) were identified as selective , orally bioavailable ChK1 inhibitors offering excellent in vitro potency with significantly reduced P22303 activity . In combination with gemcitabine , these compounds demonstrate an in vivo pharmacodynamic effect and are efficacious in a mouse p53 mutant xenograft model . [ Measurement of rifampicin and clarithromycin in serum by high-performance liquid chromatography with electrochemical detection ] . DB01045 ( RFP ) induces hepatic drug-metabolizing enzymes , making drug interactions a very important clinical problem . DB01211 ( P62158 ) metabolism is reportedly enhanced by induction of hepatic drug-metabolizing enzymes ( P08684 ) by RFP , so that the blood lend of P62158 decreases when RFP is administered concurrently . We connected an electrochemical detector to a high-performance liquid chromatograph ( HPLC ) for simple , rapid , easy measurement of blood concentrations of RFP and P62158 . Using samples of patient serum , normal serum , and reference standards , we compared HPLC by an external laboratory and the results of LC/MS/MS analysis with those of this new assay . A strong correlation was seen between our HPLC results and those of the external laboratory in RFP levels ( r=0.975 , p < 0.01 ) . A strong correlation was also seen between results we obtained for P62158 with the electrochemical detector in this assay and values measured by LC/MS/MS analysis ( r=0.995 , p < 0.01 ) . Our method enabled simple , rapid measurement of RFP and P62158 by connecting the HPLC and electrochemical detector in tandem . This system was used to modulate dosage during combined therapy with RFP and P62158 . The therapeutic effect for nontuberculous mycobacteriosis is expected to improve , and our HPLC is expected to be useful for simple , rapid , easy measurement of blood concentrations . P22303 antagonist potentiated insulin action in fed but not fasted state . The glucose disposal effect of insulin is doubled in response to a meal . This meal-induced insulin sensitization results from insulin acting on the liver , in the presence of a permissive hepatic parasympathetic feeding signal and elevated hepatic glutathione ( DB00143 ) , to release hepatic insulin-sensitizing substance ( HISS ) , a hormone that acts selectively on skeletal muscle to stimulate insulin-mediated glucose uptake . Blockade of the parasympathetic feeding signal to the liver , either through surgical denervation or atropine-mediated antagonism of hepatic muscarinic receptors , eliminates the HISS response , resulting in HISS-dependent insulin resistance ( HDIR ) and decreasing the response to insulin by approximately 55 % in the fed state . P01308 action in Sprague-Dawley rats , as determined with a rapidly sampled , transient euglycemic clamp in response to insulin ( 50 mU/kg ) , is decreased in a dose-dependent manner by atropine . In this study , we have used the ED75 atropine-induced model of HDIR . After a submaximal dose of atropine , potentiation of the remaining parasympathetic effect with the acetylcholinesterase antagonist neostigmine significantly restored postprandial insulin sensitization in a dose-dependent manner with peak effect at 0.1 microg/kg/min . DB01400 reversed the insulin resistance induced by partial fasting and partial muscarinic inhibition ( hepatic DB00143 levels are at fed levels ) , but not that induced by surgical hepatic denervation ( DB00143 normal , no nerve signal ) or 24-h fasting ( low DB00143 ) . No potentiation of the response to insulin by neostigmine occurred in normal , fed rats . The data suggest the use of either direct or indirectly acting cholinergic agonists for the treatment of impaired postprandial insulin sensitization . Unfolded protein response and activated degradative pathways regulation in Q9Y223 myopathy . Although intracellular beta amyloid ( Aβ ) accumulation is known as an early upstream event in the degenerative course of UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase ( Q9Y223 ) myopathy , the process by which Aβdeposits initiate various degradative pathways , and their relationship have not been fully clarified . We studied the possible secondary responses after amyloid beta precursor protein ( AβPP ) deposition including unfolded protein response ( UPR ) , ubiquitin proteasome system ( P08397 ) activation and its correlation with autophagy system . Eight Q9Y223 myopathy patients and five individuals with normal muscle morphology were included in this study . We performed immunofluorescence and immunoblotting to investigate the expression of AβPP , phosphorylated tau ( p-tau ) and endoplasmic reticulum molecular chaperones . Proteasome activities were measured by cleavage of fluorogenic substrates . The expression of proteasome subunits and linkers between proteasomal and autophagy systems were also evaluated by immunoblotting and relative quantitative real-time RT-PCR . Four molecular chaperones , glucose-regulated protein 94 ( P14625 ) , glucose-regulated protein 78 ( P11021 ) , calreticulin and calnexin and valosin containing protein ( P55072 ) were highly expressed in Q9Y223 myopathy . 20S proteasome subunits , three main proteasome proteolytic activities , and the factors linking P08397 and autophagy system were also increased . Our study suggests that AβPP deposition results in endoplasmic reticulum stress ( ERS ) and highly expressed P55072 deliver unfolded proteins from endoplasmic reticulum to proteosomal system which is activated in endoplasmic reticulum associated degradation ( ERAD ) in Q9Y223 myopathy . Excessive ubiquitinated unfolded proteins are exported by proteins that connect P08397 and autophagy to autophagy system , which is activated as an alternative pathway for degradation . DB00067 increases expression of UT-A1 , UT-A3 , and ER chaperone P11021 in the renal medulla of mice with a urinary concentrating defect . Activation of V2 receptors ( P30518 ) during antidiuresis increases the permeability of the inner medullary collecting duct to urea and water . Extracellular osmolality is elevated as the concentrating capacity of the kidney increases . Osmolality is known to contribute to the regulation of collecting duct water ( aquaporin-2 ; P41181 ) and urea transporter ( UT-A1 , UT-A3 ) regulation . AQP1KO mice are a concentrating mechanism knockout , a defect attributed to the loss of high interstitial osmolality . A P30518 -specific agonist , deamino-8-D-arginine vasopressin ( dDAVP ) , was infused into wild-type and AQP1KO mice for 7 days . UT-A1 mRNA and protein abundance were significantly increased in the medullas of wild-type and AQP1KO mice following dDAVP infusion . The mRNA and protein abundance of UT-A3 , the basolateral urea transporter , was significantly increased by dDAVP in both wild-type and AQP1KO mice . Semiquantitative immunoblots revealed that dDAVP infusion induced a significant increase in the medullary expression of the endoplasmic reticulum ( ER ) chaperone P11021 . Immunofluorescence studies demonstrated that P11021 expression colocalized with P41181 in principal cells of the papillary tip of the renal medulla . Using immunohistochemistry and immunogold electron microscopy , we demonstrate that vasopressin induced a marked apical targeting of P11021 in medullary principal cells . DB03904 -sensitive genes , P35638 and P18848 ( components of the ER stress pathway ) , were significantly increased in AQP1KO mice by dDAVP infusion . These findings strongly support an important role of vasopressin in the activation of an ER stress response in renal collecting duct cells , in addition to its role in activating an increase in UT-A1 and UT-A3 abundance . DB04892 . DB04892 , a derivative of physostigmine , was first described as an inhibitor of acetylcholinesterase ( P22303 ) and was shown to improve cognition in various experimental paradigms in rodents and dogs . It was clinically tested for Alzheimer 's disease , with moderate success in initial Phase II studies . DB04892 deserves attention for an additional quality of action : in addition to inhibiting P22303 , it modulates the amount of beta-amyloid precursor protein ( P05067 ) in neuronal cell culture by reducing P05067 translation . This effect probably involves interaction of phenserine with a regulatory element in the 5'-untranslated region of the P05067 gene that controls P05067 expression . DB04892 apparently reduces translational efficiency of P05067 mRNA into protein , a process that may involve an interaction with iron and/or an iron-responsive element . As a consequence , phenserine reduces beta-amyloid peptide ( Abeta ) formation in vitro and in vivo . DB04892 is also unique because of differing actions of its enantiomers : DB04892 is the active enantiomer for inhibition of P22303 , whereas (+)-phenserine ( ' posiphen ' ) has weak activity as an P22303 inhibitor and can be dosed much higher . Both enantiomers are equipotent in downregulating P05067 expression . (+)-Posiphen may be a promising drug , either alone or in combination with DB04892 , to attenuate the progression of Alzheimer 's disease . DB06403 . Elevated endothelin ( ET ) -1 levels are strongly correlated with the pathogenesis and prognosis of pulmonary arterial hypertension ( PAH ) . DB06403 is an orally active , highly selective P25101 receptor antagonist with > 4000-fold higher selectivity over the ETB receptor . In two large , well designed , 12-week , placebo-controlled , phase III trials ( ARIES-1 , n = 202 and ARIES-2 , n = 192 ) in patients with PAH ( WHO group I ) , ambrisentan 2.5-10 mg once daily significantly increased 6-minute walk distance by 31-59 m from baseline ( primary outcome measure ) versus placebo . The incidence of clinical worsening ( secondary outcome measure ) was significantly delayed for the combined ambrisentan 5 mg once daily groups versus the combined placebo groups from ARIES-1 and -2 . At week 12 , WHO functional class distribution was significantly improved with once-daily ambrisentan 5 mg , and Borg dyspnoea scores were significantly improved with ambrisentan 2.5-10 mg versus placebo in combined data from the ARIES-1 and -2 trials . The beneficial effects of ambrisentan on exercise capacity , WHO functional class and Borg dyspnoea scores seen at 12 weeks were maintained at 48 weeks in the ARIES-E phase III extension trial ( n = 361 ) . One-year survival rates with ambrisentan were 95-97 % . Treatment with ambrisentan for up to 2.8 years was generally well tolerated in clinical trials . DB01411 inhibits NF-kappaB activation and Q02817 gene expression in cultured human epithelial cells . DB01411 is a selective cysteinyl leukotriene ( 1 ) (cysLT(1)) receptor antagonist , and is now widely used in the treatment of asthma . The anti-asthmatic effect of pranlukast may be rendered not only by antileukotriene activity , but also by other pharmacological activity . This study was designed to investigate whether pranlukast had inhibitory effects on nuclear factor-kappaB ( NF-kappaB ) activation and mucin gene expression in cultured human epithelial cells . Luciferase assay was mainly used for analysis . Cultured epithelial cells were transfected with NF-kappaB luciferase vector , Q02817 or P98088 luciferase vectors . Lipopolysaccharide ( LPS ) significantly increased NF-kappaB activation in NCI-H292 cells , which was inhibited by the pretreatment by pranlukast in a dose-dependent manner . Either LTD(4) or pranlukast alone did not increase NF-kappaB activation in NCI-H292 cells . DB01411 also inhibited NF-kappaB activation induced by phorbol 12-myristate 13-acetate ( PMA ) . DB01411 also significantly inhibited LPS-induced Q02817 mRNA expression by reverse transcription-polymerase chain reaction ( RT-PCR ) analysis in NCI-H292 cells . DB01411 also inhibited LPS-induced Q02817 gene expression in HM3- Q02817 cells . However , pranlukast did not inhibit P98088 gene transcription activity induced by lipoteichoic acid ( P01374 ) in NCI-H292 cells . These results suggest that pranlukast may inhibit NF-kappaB activation and Q02817 gene transcription through pathways distinct from cysLT(1) receptor antagonism in cultured human epithelial cells . Mechanisms underlying hypertriglyceridemia in rats with monosodium L-glutamate-induced obesity : evidence of P17861 / P07237 / P55157 axis activation . Non-alcoholic fatty liver disease ( NAFLD ) is intimately associated with insulin resistance and hypertriglyceridemia , whereas many of the mechanisms underlying this association are still poorly understood . In the present study , we investigated the relationship between microsomal triglyceride transfer protein ( P55157 ) and markers of endoplasmic reticulum ( ER ) stress in the liver of rats subjected to neonatal monosodium L-glutamate ( MSG ) -induced obesity . At age 120 days old , the MSG-obese animals exhibited hyperglycemia , hypertriglyceridemia , insulin resistance , and liver steatosis , while the control ( CTR ) group did not . Analysis using fast protein liquid chromatography of the serum lipoproteins revealed that the triacylglycerol content of the very low-density lipoprotein ( VLDL ) particles was twice as high in the MSG animals compared with the CTR animals . The expression of ER stress markers , GRP76 and P14625 , was increased in the MSG rats , promoting a higher expression of X-box binding protein 1 ( P17861 ) , protein disulfide isomerase ( P07237 ) , and P55157 . As the P17861 / P07237 / P55157 axis has been suggested to represent a significant lipogenic mechanism in the liver response to ER stress , our data indicate that hypertriglyceridemia and liver steatosis occurring in the MSG rats are associated with increased P55157 expression . The Drosophila insulin receptor activates multiple signaling pathways but requires insulin receptor substrate proteins for DNA synthesis . The Drosophila insulin receptor ( P30518 ) contains a 368-amino-acid COOH-terminal extension that contains several tyrosine phosphorylation sites in YXXM motifs . This extension is absent from the human insulin receptor but resembles a region in insulin receptor substrate ( P41252 ) proteins which binds to the phosphatidylinositol ( PI ) 3-kinase and mediates mitogenesis . The function of a chimeric P30518 containing the human insulin receptor binding domain ( hDIR ) was investigated in 32D cells , which contain few insulin receptors and no P41252 proteins . P01308 stimulated tyrosine autophosphorylation of the human insulin receptor and hDIR , and both receptors mediated tyrosine phosphorylation of Shc and activated mitogen-activated protein kinase . P35568 was required by the human insulin receptor to activate PI 3-kinase and p70s6k , whereas hDIR associated with PI 3-kinase and activated p70s6k without P35568 . However , both receptors required P35568 to mediate insulin-stimulated mitogenesis . These data demonstrate that the P30518 possesses additional signaling capabilities compared with its mammalian counterpart but still requires P35568 for the complete insulin response in mammalian cells . Beyond statins : new lipid lowering strategies to reduce cardiovascular risk . Statins are the first-line therapy in LDL- DB04540 ( LDL-C ) reduction and its clinical use has contributed to significant prevention and treatment of atherosclerotic vascular disease . Yet , a significant proportion of patients remain at high risk . Recently , a number of new therapies have been developed to further lower LDL-C . These agents may provide clinical benefit on top of statin therapy in patients with high residual risk , severe hypercholesterolemia or as an alternative for patients who are intolerant to statins . We review four novel approaches based on the inhibition of proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) , apolipoprotein-B100 ( apoB ) , Cholesteryl ester transport protein ( P11597 ) and microsomal triglyceride transfer protein ( P55157 ) . ApoB and P55157 inhibitors ( DB05528 and DB08827 ) are indicated only for homozygous familial hypercholesterolemia patients . The results of ongoing trials with P11597 and Q8NBP7 inhibitors may warrant a wider employment in different categories of patients at high risk for cardiovascular disease . DB08827 : A novel agent for the treatment of homozygous familial hypercholesterolemia . PURPOSE : The pharmacology , pharmacokinetics , and clinical efficacy and safety of lomitapide in the management of homozygous familial hypercholesterolemia ( HoFH ) are reviewed . SUMMARY : DB08827 ( Juxtapid , Aegerion Pharmaceuticals ) is an oral microsomal triglyceride transfer protein ( P55157 ) inhibitor indicated for the treatment of patients with HoFH , a rare form of hypercholesterolemia that can lead to premature atherosclerotic disease . In clinical trials , the use of lomitapide alone or in combination with other lipid-lowering modalities reduced plasma concentrations of low-density lipoprotein cholesterol ( LDL-C ) by a mean of more than 50 % . DB08827 is associated with significant gastrointestinal adverse effects and increases in hepatic fat levels . DB08827 undergoes hepatic metabolism via cytochrome P-450 ( CYP ) isoenzyme 3A4 and interacts with P08684 substrates including atorvastatin and simvastatin ; dose adjustment is recommended when lomitapide is used concurrently with these agents . In patients receiving concomitant warfarin , the International Normalized Ratio ( INR ) should be closely monitored , as lomitapide use may increase INR values . The recommended initial dosage of lomitapide is 5 mg once daily , with subsequent upward dose adjustment at specified intervals according to tolerability . DB08827 is contraindicated in patients with moderate-to-severe liver disease , patients with sustained abnormal liver function tests , patients taking strong or moderate P08684 inhibitors , and pregnant patients . CONCLUSION : DB08827 is an oral P55157 inhibitor approved for the treatment of HoFH . This agent appears to be a realistic option for patients with HoFH who are unable to attain their LDL-C goal or can not tolerate statin therapy . DB08875 suppresses tumor growth and metastasis in hepatocellular carcinoma by a dual blockade of P35968 and MET . PURPOSE : MET signaling has been suggested a potential role in hepatocellular carcinoma ( HCC ) and associated with prometastasis during antiangiogenesis therapy . We investigated the potential association between MET expression and therapeutic response to sorafenib in patients with HCC . Antitumor effects of cabozantinib , a dual inhibitor of MET and P35968 , were examined in cultured HCC cells as well as in vivo models . EXPERIMENTAL DESIGN : Total MET and phosphorylated MET ( p-MET ) were measured in 29 resected HCC specimens , and correlated with response to sorafenib as postoperative adjuvant therapy . In the second set of experiments using cultured HCC cells , and mouse xenograft and metastatic models , effects of cabozantinib were examined . RESULTS : High level of p-MET in resected HCC specimens was associated with resistance to adjuvant sorafenib therapy . In cultured HCC cells that expressed p-MET , cabozantinib inhibited the activity of MET and its downstream effectors , leading to P55008 -phase arrest . DB08875 inhibited tumor growth in p-MET-positive and p-MET-negative HCC by decreasing angiogenesis , inhibiting proliferation , and promoting apoptosis , but it exhibited more profound efficacy in p-MET-positive HCC xenografts . DB08875 blocked the hepatocyte growth factor ( P14210 ) -stimulated MET pathway and inhibited the migration and invasion of the HCC cells . Notably , cabozantinib reduced the number of metastatic lesions in the lung and liver in the experimental metastatic mouse model . CONCLUSIONS : Patients with HCC with high level of p-MET are associated with resistance to adjuvant sorafenib treatment . The dual blockade of P35968 and MET by cabozantinib has significant antitumor activities in HCC , and the activation of MET in HCC may be a promising efficacy-predicting biomarker . Clin Cancer Res ; 20(11) ; 2959-70 . ©2014 AACR . DB00428 -induced increase in cholesterol ester transfer protein ( P11597 ) and its reversal by insulin in transgenic mice expressing human P11597 . High plasma triacylglycerol and low high-density lipoprotein levels are risk factors for cardiovascular disease in diabetes . Plasma high-density lipoprotein levels are regulated by cholesterol ester transfer protein ( P11597 ) . The regulation of P11597 under diabetic conditions is not clear , and this is due to a lack of appropriate models . We used transgenic mice expressing human P11597 to study the regulation of this protein under type-1 diabetic conditions and further investigated whether insulin reverses the effect of diabetes . Mice expressing human P11597 under the control of its natural flanking region and age-matched littermates not expressing this protein were made diabetic by injecting streptozotocin , and the reversal of diabetes was assessed by injecting insulin . The plasma total cholesterol , low-density lipoprotein-cholesterol , and triacylglycerol concentrations were elevated , whereas high-density lipoprotein-cholesterol concentrations were reduced after the onset of diabetes . P01308 injection partially recovered this effect . The plasma cholesterol ester transfer activity , P11597 mass , and hepatic P11597 mRNA abundance were significantly higher in diabetic mice that were partially restored by insulin administration . There was a strong correlation between high-density lipoprotein-cholesterol concentrations and cholesterol ester transfer activity . These results suggest that an increase in P11597 under diabetic conditions might be a major factor responsible for increased incidence of diabetes-induced atherosclerosis . Nitrergic response to cyclophosphamide treatment in blood and bone marrow . Daily intraperitoneal injection of cyclophosphamide ( P15085 ) ( 50 mgkg(-1) of body weight ) for 5 days resulted in reduced levels of marrow and blood cellularity , which was most pronounced in 18 days post-treatment ( pt ) . On day 18 after P15085 treatment the enhancedlevels of nitric oxide ( NO ) precursors and metabolites ( L-arginine , L-citrulline , reactive nitrogen species ( RNS ) ) of marrow and blood cells ( platelet , neutrophil , lymphocyte and monocyte ) resulted from up-regulation of Ca(II)/calmodulin( P62158 )-independent " inducible " NO synthase ( P35228 ) , with a lessercontribution of Ca(II)/ P62158 -dependent " constitutive " P29474 isoforms to systemic NO.Biphasic response to P15085 of marrow nitrergic system , i.e. both P35228 and P29474 showed significantly depressed activities , as well as diminished levels of NO metabolites on day 9 pt , suggested that signals in addition to NO might be involved in P15085 -induced inhibition of hematopoesis , while a gradual increase of neutrophil and platelet NOS activity appeared to be contributed to a P15085 -induced development of granulopenia , thrombocytopenia and hemorrhage . Systemic sclerosis - a systematic overview : part 2 - immunosuppression , treatment of SSc-associated vasculopathy , and treatment of pulmonary arterial hypertension . Here we give an overview over treatment recommendations propagated by the European League Against Rheumatism ( EULAR ) , EULAR Scleroderma Trials and Research Group , the German Network for Systemic Sclerosis , the European Respiratory Society , and the International Society of Heart and Lung Transplantation . As response to immunosuppressant ( IS ) therapy is usually weaker in systematic sclerosis ( SSc ) compared to other connective tissue disorders IS should be considered with caution . To prevent scleroderma renal crisis steroid doses should not exceed 15 mg/d . The definitive role of a number of new immunosuppressant drugs and the effects of autologous stem cell transplantation in systemic clerosis ( SSc ) have to be elucidated . Prostanoids , especially iloprost , are widely used as intravenous formulas for the treatment of severe Raynaud 's phenomenon ( RP ) and digital ulcers ( DU ) . DB01373 antagonists are of limited therapeutic value . DB00559 , an oral endothelin receptor antagonists ( P25101 ) , was shown to prevent new DU , but failed to heal existing DU , while the oral phopshodiesterase inhibitor ( P07237 ) DB00203 reduces the occurrence of RP and might be effective in ulcer healing . Combination therapies of P07237 with P25101 are currently evaluated . Therapy of pulmonary arterial hypertension ( PAH ) is usually started as oral monotherapy , frequently using an P25101 . When this first-line therapy is not tolerated P25101 is substituted by P07237 . If treatment goals are not reached with monotherapy combinationtherapy is started , for example by adding a P07237 to an existing P25101 . In general , treatment of PAH in patients with connective tissue disease follows the same algorithms as in idiopathic PAH . A common binding site on the microsomal triglyceride transfer protein for apolipoprotein B and protein disulfide isomerase . The assembly of triglyceride-rich lipoproteins requires the formation in the endoplasmic reticulum of a complex between apolipoprotein B ( apoB ) , a microsomal triglyceride transfer protein ( P55157 ) , and protein disulfide isomerase ( P07237 ) . In the P55157 complex , the amino-terminal region of P55157 ( residues 22-303 ) interacts with the amino-terminal region of apoB ( residues 1-264 ) . Here , we report the identification and characterization of a site on apoB between residues 512 and 721 , which interacts with residues 517-603 of P55157 . P07237 binds in close proximity to this apoB binding site on P55157 . The proximity of these binding sites on P55157 for P07237 and amino acids 512-721 of apoB was evident from studies carried out in a yeast two-hybrid system and by co-immunoprecipitation . The expression of P07237 with P55157 and apoB16 ( residues 1-721 ) in the baculovirus expression system reduced the amount of P55157 co-immunoprecipitated with apoB by 73 % . The interaction of residues 512-721 of apoB with P55157 facilitates lipoprotein production . Mutations of apoB that markedly reduced this interaction also reduced the level of apoB-containing lipoprotein secretion . Imatinib inhibits P15692 -independent angiogenesis by targeting neuropilin 1-dependent P00519 activation in endothelial cells . To enable new blood vessel growth , endothelial cells ( ECs ) express neuropilin 1 ( NRP1 ) , and NRP1 associates with the receptor tyrosine kinase P35968 after binding the vascular endothelial growth factor A ( P15692 ) to enhance arteriogenesis . We report that NRP1 contributes to angiogenesis through a novel mechanism . In human and mouse ECs , the integrin ligand fibronectin ( FN ) stimulated actin remodeling and phosphorylation of the focal adhesion component paxillin ( P49023 ) in a P15692 / P35968 -independent but NRP1-dependent manner . NRP1 formed a complex with P00519 that was responsible for FN-dependent P49023 activation and actin remodeling . This complex promoted EC motility in vitro and during angiogenesis on FN substrates in vivo . Accordingly , both physiological and pathological angiogenesis in the retina were inhibited by treatment with Imatinib , a small molecule inhibitor of P00519 which is widely used to prevent the proliferation of tumor cells that express P11274 - P00519 fusion proteins . The finding that NRP1 regulates angiogenesis in a P15692 - and P35968 -independent fashion via P00519 suggests that P00519 inhibition provides a novel opportunity for anti-angiogenic therapy to complement P15692 or P35968 blockade in eye disease or solid tumor growth . Consequences of missense mutations for dimerization and turnover of alanine:glyoxylate aminotransferase : study of a spectrum of mutations . DB00160 :glyoxylate aminotransferase ( AGT ) is a liver peroxisomal enzyme , deficiency of which results in primary hyperoxaluria type 1 ( P78364 ) . More than 65 P78364 -related mutations are now documented in the AGT gene ( P21549 ) , of which about 50 % are missense . We have generated a spectrum of 15 missense changes including the most common P78364 mutation , G170R , and expressed them on the appropriate background of the major or minor allele , in an Escherichia coli overexpression system and in a rabbit reticulocyte transcription/translation system . We have investigated their effects on enzyme activity , dimerization , aggregation , and turnover . The effect of pyridoxal phosphate ( PLP ) on dimerization and stability was also investigated . Although all 15 mutant AGTs were expressed as intact proteins in E. coli , only three : G41R and G41V on the major allele , and the common mutation G170R , resulted in significant amounts of enzymatic activity . Dimerization failure was a frequent observation ( 13/15 ) except for G41V and D183N . Dimerization was poor with S187F but was substantially improved with PLP . Proteasome-mediated protein degradation was observed for all the mutations except G41R on the major allele , G41V , D183N , G170R , and S218L . Increases in the stability of the mutant enzymes in the presence of PLP were small ; however , G41R on the minor allele showed a direct relationship between its half life and the concentration of PLP . The minor allele AGT product and many of the mutants were subject to a limited non-proteasomal proteolytic cleavage when DB00171 was depleted . Endoplasmic reticulum stress in PLP-overexpressing transgenic rats : gray matter oligodendrocytes are more vulnerable than white matter oligodendrocytes . Studies dealing with transport of proteins from the oligodendrocyte cell body to the myelin sheath reveal the presence of different transport pathways . Proteolipid protein ( PLP ) is synthesized at the rough endoplasmic reticulum ( ER ) and then processed through the Golgi apparatus and transported to the myelin membranes . P02686 ( MBP ) on the other hand is synthesized locally at the ends of cell processes where its messenger RNA is translated on free ribosomes . Here we show that in rats that overexpress PLP , impairment of PLP transport from the cell body to the processes interferes with the translocation of other membrane proteins such as myelin-associated glycoprotein ( P20916 ) and myelin oligodendrocyte glycoprotein ( Q16653 ) , but not with peripherally translated MBP . In addition , it also impedes the transport of non-myelin proteins , for example the amyloid precursor protein ( P05067 ) . At the ultrastructural level , the ER of these metabolically disturbed oligodendrocytes revealed extreme swelling of the cisternae , and immunohistochemistry revealed intense expression of the ER chaperone molecule P11021 / P11021 and ER folding enzyme protein disulfide isomerase ( P07237 ) . These features suggest that these oligodendrocytes , which were found exclusively in gray matter areas of the spinal cord , started an unfolded protein response while suffering from ER stress . Some of these disturbed oligodendrocytes were seen to undergo programmed cell death . These results indicate that gray matter oligodendrocyte differ from white matter oligodendrocytes in their capacity to stabilize metabolic disturbances by an unfolded protein response . P29474 activation by HDL is impaired in genetic P11597 deficiency . Mutations in the P11597 gene resulting in defective P11597 activity have been shown to cause remarkable elevations of plasma HDL-C levels , with the accumulation in plasma of large , buoyant HDL particles enriched in apolipoprotein E. Genetic P11597 deficiency thus represents a unique tool to evaluate how structural alterations of HDL impact on HDL atheroprotective functions . Aim of the present study was to assess the ability of HDL obtained from P11597 -deficient subjects to protect endothelial cells from the development of endothelial dysfunction . HDL isolated from one homozygous and seven heterozygous carriers of P11597 null mutations were evaluated for their ability to down-regulate cytokine-induced cell adhesion molecule expression and to promote NO production in cultured endothelial cells . When compared at the same protein concentration , HDL and HDL3 from carriers proved to be as effective as control HDL and HDL3 in down-regulating cytokine-induced P19320 , while carrier HDL2 were more effective than control HDL2 in inhibiting P19320 expression . On the other hand , HDL and HDL fractions from carriers of P11597 deficiency were significantly less effective than control HDL and HDL fractions in stimulating NO production , due to a reduced P29474 activating capacity , likely because of a reduced Q14703 content . In conclusion , the present findings support the notion that genetic P11597 deficiency , by affecting HDL particle structure , impacts on HDL vasculoprotective functions . Understanding of these effects might be important for predicting the outcomes of pharmacological P11597 inhibition .

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Prevention of acute and chronic allograft rejection by a novel retinoic acid receptor-alpha-selective agonist . To investigate the involvement of retinoic acid receptor ( RAR ) -alpha in allograft rejection , we investigated the effect of a novel selective agonist to the receptor , ER-38925 , in a mouse cardiac allograft model . Prophylactic treatment with ER-38925 inhibited the acute rejection of the mouse cardiac allograft ( BALB/c --> C3H/HeN ) at 0.3 and 3 mg/kg , and its effect was enhanced in combination with tacrolimus . In this model , ER-38925 remarkably inhibited cytotoxic T lymphocyte induction and alloantigen-stimulated production of cytokines , i.e. P60568 , IL-12 and P01579 . In the chronic rejection model , combined treatment with tacrolimus and ER-38925 reduced the grade and incidence of arteriosclerosis in the cardiac allografts significantly more potently than tacrolimus monotherapy . ER-38925 inhibited the proliferation of rat aortic smooth muscle cells stimulated in vitro , presumably through the induction of a cyclin-dependent kinase inhibitor , p27(kip-1) . Those results provide a rationale for using P10276 agonists as immunosuppressants in human organ transplantation . Synthesis and biological evaluation of novel ( 4 or 5-aryl ) pyrazolyl-indoles as inhibitors of interleukin-2 inducible T-cell kinase ( Q08881 ) . P60568 inducible T-cell kinase ( Q08881 ) is one of five kinases that belong to the Tec kinase family that plays an important role in T-cell and mast cell signaling . Various reports point to a role of Q08881 in the treatment of allergic asthma . For example , it was shown that mice lacking Q08881 have reduced airway hyperresponsiveness , inflammation and tracheal responses in an allergic asthma model . In this article , we disclose novel Q08881 inhibitors based on ( 4 or 5-aryl ) pyrazolyl-indole scaffold that were also found to be selective for Q08881 over other kinases like IRK , P24941 , GSK3ss and PKA . Growth-inhibitory effects of vitamin D analogues and retinoids on human pancreatic cancer cells . Retinoids and vitamin D are important factors that regulate cellular growth and differentiation . An additive growth-inhibitory effect of retinoids and vitamin D analogues has been demonstrated for human myeloma , leukaemic and breast cancer cells . We set out to study the effects of the vitamin D analogue EB1089 and the retinoids all-trans- and 9-cis-retinoic acid on the human pancreatic adenocarcinoma cell lines Capan 1 and Capan 2 and the undifferentiated pancreatic carcinoma cell line Hs766T . The cell lines investigated expressed vitamin D receptor , retinoic acid receptor ( RAR ) -alpha and gamma as determined by polymerase chain reaction after reverse transcription . P10826 was expressed only in Hs766T cells . Addition of all-trans-retinoic acid increased the amount of P10276 mRNA in the three cell lines and induced P10826 mRNA in Capan 1 and Capan 2 cells . All-trans-retinoic acid at a concentration of 10 nM inhibited the growth of Capan 1 and Capan 2 cells by 40 % relative to controls . DB00523 was less effective . Neither all-trans-retinoic acid nor 9-cis-retinoic acid affected the growth of Hs766T cells . EB1089 , if added alone to the cells , did not significantly inhibit growth . However , the combination of 1 nM EB1089 with 10 nM all-trans-retinoic acid exerted a growth-inhibitory effect of 90 % in Capan 1 cells and of 70 % in Capan 2 cells . Our data suggest that vitamin D analogues together with retinoids inhibit the growth of human pancreatic cancer cells . However , in vivo studies are necessary to examine the potential use of retinoids and vitamin D analogues on pancreatic cancer . Effect of 5- Q9H205 receptor antagonist ( ondansetron ) on functioning human pancreatic carcinoid cells . 5-Hydroxytryptamine ( 5-HT ) is a mitogen for selected cell types . We have reported that 5-HT is an autocrine growth factor for functioning human pancreatic carcinoid ( BON ) cells ; autocrine growth effect is transmitted by P08908 but not P28335 /2 receptors , activation of which decreases cyclic AMP production through a pertussis toxin-sensitive inhibitory GTP-binding protein . In this study , the effect of 5- Q9H205 receptor antagonist , ondansetron , on BON was examined . DB00904 did not affect growth of BON cells and also affected neither stimulation of phosphatidylinositol hydrolysis or inhibition of cyclic AMP production evoked by 5-HT in BON cells . DB00904 , however , inhibited mobilization of intracellular calcium evoked by 5-HT . Present findings suggest that BON cells possess 5- Q9H205 receptors , but their roles in pancreatic carcinoid cells are still unknown . Displacement of Q01826 -bound histone deacetylase 1 corepressor by the human immunodeficiency virus type 1 transactivator induces expression of interleukin-2 and its receptor in T cells . One hallmark of human immunodeficiency virus type 1 ( HIV-1 ) infection is the dysregulation of cytokine gene expression in T cells . Transfection of T cells with human T-cell leukemia type 1 or 2 transactivator results in the induction of the T-cell-restricted cytokine interleukin-2 ( P60568 ) and its receptor ( IL-2Ralpha ) . However , no T-cell-specific factor(s) has been directly linked with the regulation of P60568 and IL-2Ralpha transcription by influencing the promoter activity . Thymocytes from Q01826 ( special AT-rich sequence binding protein 1 ) knockout mice have been shown to ectopically express IL-2Ralpha , suggesting involvement of Q01826 in its negative regulation . Here we show that Q01826 , a T-cell-specific global gene regulator , binds to the promoters of human P60568 and IL-2Ralpha and recruits histone deacetylase 1 ( Q13547 ) in vivo . Q01826 also interacts with Tat in HIV-1-infected T cells . The functional interaction between HIV-1 Tat and Q01826 requires its PDZ-like domain , and the binding of the Q13547 corepressor occurs through the same . Furthermore , Tat competitively displaces Q13547 that is bound to Q01826 , leading to increased acetylation of the promoters in vivo . Transduction with Q01826 interaction-deficient soluble Tat ( Tat 40-72 ) and reporter assays using a transactivation-negative mutant ( C22G ) of Tat unequivocally demonstrated that the displacement of Q13547 itself is sufficient for derepression of these promoters in vivo . These results suggest a novel mechanism by which HIV-1 Tat might overcome Q01826 -mediated repression in T cells . Predictive model for risk of severe gastrointestinal toxicity following chemotherapy using patient immune genetics and type of cancer : a pilot study . PURPOSE : Severe chemotherapy-induced gastrointestinal toxicity ( CIGT ) is common and results in treatment delays , dose reductions , and potential premature treatment discontinuation . Currently , there is no diagnostic marker to predict CIGT . Proinflammatory cytokines , produced via toll-like receptor signaling , are key mediators of this toxicity . Hence , this pilot study investigated the association between immune genetic variability and severe CIGT risk . METHODS : Genomic DNA from 34 patients ( 10 with severe CIGT ) who had received 5-fluoruracil-based chemotherapy regimens was analyzed for variants of IL-1B , P60568 , P05231 , IL-6R , P22301 , P01375 , TGF-b , O60603 , O00206 , Q9Y6Y9 , Q99836 , P23560 , CRP , ICE , and P35372 . Multivariate logistic regression created a prediction model of severe CIGT risk . RESULTS : There were no significant differences between patients with and without severe CIGT with regards to age , sex , type of cancer , or chemotherapy treatment regimens . The prediction model of severe CIGT risk included O60603 and P01375 genetic variability and cancer type ( colorectal and gastric ) . This prediction model was both specific and sensitive , with a receiver operator characteristic area under the curve of 87.3 % . CONCLUSIONS : This is the first report of immune genetic variability , together with cancer type , being predictive of severe CIGT risk . These outcomes are being validated in a larger patient population . DB06594 targets a range of major depressive disorder symptoms . Servier , and US licensee Novartis AG , are developing the oral melatonin MT1 and P02795 agonist and P41595 and P28335 antagonist agomelatine as a once-daily treatment for major depressive disorder ( MDD ) and its symptoms , particularly anxiety , and sleep and circadian disturbances . Phase III trials have been completed and a registration dossier has been submitted to the EMEA in Western Europe . Regulation of Con A-dependent cytokine production from P01730 + and CD8+ T lymphocytes by autosecretion of histamine . OBJECTIVES : Previously we have shown that both P01730 + T cells and CD8+ T cells produce histamine when activated with Con A . The aim of this study was to examine whether cytokine production by these cells is regulated by autosecretion of histamine . MATERIALS : P01730 + and CD8+ T cells were separated from spleen cells of C57BL/6 mice and mice lacking the H1 receptor ( P35367 ) or P25021 , using anti- P01730 +- and anti-CD8+-coupled magnetic beads , respectively . RESULTS : Depletion of the P35367 resulted in decreases in the release of P60568 and P22301 from both P01730 + and CD8+ cells and increases in the release of P05112 from P01730 + T cells and P01579 from CD8+ cells . Mice lacking the P25021 showed up-regulation of P01579 secretion from CD8+ cells and of P05112 from P01730 + and CD8+ T cells . Release of P60568 and P22301 from P01730 + as well as CD8+ cells was down-regulated in these mice . Both P01730 + and CD8+ T cell fractions synthesized histamine , which was enhanced in the P35367 -deficient CD8+ T cells . Treatment of the cells with alpha-fluoromethyl-histidine , a specific inhibitor of HDC , or histaminase increased P01579 from CD8+ cells , whereas it had no appreciable effect on P05112 secretion from P01730 + cells . CONCLUSION : These results suggest that cytokine production by P01730 + and CD8+ T lymphocytes is regulated by autosecretion of histamine . Discovery of ( 2E ) -3-{2-butyl-1-[2-(diethylamino)ethyl]-1H-benzimidazol-5-yl}-N-hydroxyacrylamide ( DB05223 ) , an orally active histone deacetylase inhibitor with a superior preclinical profile . A series of 3-(1,2-disubstituted-1H-benzimidazol-5-yl)-N-hydroxyacrylamides ( 1 ) were designed and synthesized as HDAC inhibitors . Extensive SARs have been established for in vitro potency ( Q13547 enzyme and COLO 205 cellular IC(50) ) , liver microsomal stability ( t(1/2) ) , cytochrome P450 inhibitory ( 3A4 IC(50) ) , and clogP , among others . These parameters were fine-tuned by carefully adjusting the substituents at positions 1 and 2 of the benzimidazole ring . After comprehensive in vitro and in vivo profiling of the selected compounds , DB05223 ( 3 ) was identified as a preclinical development candidate . 3 is a potent pan-HDAC inhibitor with excellent druglike properties , is highly efficacious in in vivo tumor models ( HCT-116 , PC-3 , A2780 , MV4-11 , Ramos ) , and has high and dose-proportional oral exposures and very good ADME , safety , and pharmaceutical properties . When orally dosed to tumor-bearing mice , 3 is enriched in tumor tissue which may contribute to its potent antitumor activity and prolonged duration of action . 3 is currently being tested in phase I and phase II clinical trials . Functional significance of DB00142 -77 and DB00135 -137 within the active site of isoaspartyl dipeptidase . Q7L266 ( IAD ) is a binuclear metalloenzyme and a member of the amidohydrolase superfamily . This enzyme catalyzes the hydrolytic cleavage of beta-aspartyl dipeptides . The pH-rate profiles for the hydrolysis of beta- DB00128 - DB00149 indicates that catalysis is dependent on the ionization of two groups ; one that ionizes at a pH approximately 6 and the other approximately 9 . The group that must be ionized for catalysis is directly dependent on the identity of the metal ion bound to the active site . This result is consistent with the ionization of the hydroxide that bridges the two divalent cations . In addition to the residues that interact directly with the divalent cations there are two other residues that are highly conserved and found within the active site : DB00142 -77 and DB00135 -137 . Mutation of DB00135 -137 to phenylalanine reduced the rate of catalysis by three orders of magnitude . The three dimensional X-ray structure of the Y137F mutant did not show any significant conformation changes relative to the three dimensional structure of the wild-type enzyme . The positioning of the side-chain phenolic group of DB00135 -137 in the active site of IAD is consistent with the stabilization of the tetrahedral adduct concomitant with nucleophilic attack by the hydroxide that bridges the two divalent cations . Mutation of DB00142 -77 resulted in the reduction of catalytic activity by five orders of magnitude . The three dimensional structure of the E77Q mutant did not show any significant conformational changes in the mutant relative to the three dimensional structure of the wild-type enzyme . The positioning of the side-chain carboxylate of DB00142 -77 is consistent with the formation of an ion pair interaction with the free alpha-amino group of the substrate . Cholecystokinin-related peptides , after systemic or central administration , prevent carbon monoxide-induced amnesia in mice . The neuroprotective actions of cholecystokinin ( CCK ) peptides were investigated in a mouse hypoxia model , in which the animals were successively exposed to CO gas . Working memory impairment 5 days after CO exposure was examined by using a Y-maze test ; delayed amnesia was examined 7 days after CO exposure , by using a step-down type passive avoidance test . DB00403 ( 1-100 micrograms/kg , given s.c. 30 min before CO exposure ) significantly prevented the CO-induced impairment of performance in both tests , the improvement being correlated with the severity of hypoxia . This severity was increased by maintaining the body temperature at 38 degrees C . DB00403 was less effective when injected immediately after a single CO exposure . The order of potency of the CCK-peptides administered systemically was : ceruletide > CCK-8S > CCK-8NS >> Q13308 . DB00403 ( 0.03-0.3 micrograms/mouse ) and CCK-8S ( 0.03-1 microgram/mouse ) prevented CO-induced amnesia after i.c.v. administration . Under all experimental conditions , dizocilpine [ MK-801 , (+)-5-methyl-10,11-dihydro-5H-dibenzo(a,d)cyclohepten-5,10-imine maleate , 500 micrograms/kg s.c. or 10 micrograms/mouse i.c.v. ] prevented completely the CO-induced amnesia . The protective effects of systemic ceruletide were blocked , partially but significantly , by the preadministration of L-364,718 ( 3S-(-)-N- [ 2,3-dihydro-1-methyl-2-oxo-S-phenyl-1H-1,4- benzodiazepine-3-yl ] -1H-indole-2-carboxamide , 1-10 mg/kg i.p. ) , a selective P32238 antagonist . L-365,260 ( [ 3R-(+)-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepine-3-yl ] -N ' - [3-methyl-phenyl]urea ) , a CCK-B antagonist , also decreased ceruletide-induced protection. ( ABSTRACT TRUNCATED AT 250 WORDS ) DB08890 - a secretagogue and antihyperalgesic agent - what next ? Ongoing clinical trials suggest that linaclotide , a first-in-class , 14-amino acid peptide guanylate cyclase-C ( P25092 ) receptor agonist and intestinal secretagogue is an effective treatment for chronic constipation . A study in this issue of the Journal suggests that linaclotide also has antihyperalgesic effects in three common rat models of inflammation- and stress-induced hypersensitivity ( i.e. , acute trinitrobenzene sulfonic acid colitis , water avoidance stress [ P42768 ] , and restraint-induced stress ) but not in naïve animals . In mice , linaclotide at least partly reduces hyperalgesia via P25092 receptors . Dose-effect relationships of linaclotide were complicated and non-linear . This viewpoint discusses human clinical trials with linaclotide and the results of this study . Potential mechanisms and clinical significance of these findings are explored . Collectively , these data suggest that P25092 receptors exert other , as yet poorly understood , effects on gastrointestinal sensitivity in conditions associated with inflammation and/or stress-induced increased intestinal permeability . However , the data need to be confirmed in humans and in long-term animal models . Further studies are also necessary to elucidate the mechanisms as these effects can not be explained by linaclotide 's known effects on epithelial P25092 receptors . DB00075 -induced anergy in cloned human Th0 , Th1 , and Th2 cells . In the mouse , activation of T cells by T cell receptor ( TCR ) crosslinking with anti-CD3 antibodies in the absence of a costimulatory signal induces Th1 but not Th2 cell anergy . Furthermore , anti-CD3 induces anergy of Th1- but not Th2-type lymphokine secretion in Th0 cells . This study was designed to determine whether this is also the case in man . Human rye grass allergen Lol p I-specific cloned P01730 + T helper cells of subtypes Th0 , Th1 , and Th2 were treated with immobilized anti-CD3 . The cells were rested for 4 days and then activated under optimal conditions with antigen and antigen-presenting cells ( APCs ) . Cell proliferation and P60568 , P01579 , and P05112 secretion was determined to test for the anergic state . The initial anti-CD3 treatment induced cell proliferation , P60568 , P01579 , and/or P05112 secretion by T cells of all three subsets which was followed by an anergic state in Th0 , Th1 , and Th2 cells as shown by a 51 to > 94 % decrease in cell proliferation and P01579 and/or P05112 secretion after subsequent P25054 and Lol p I activation . Addition of P60568 or P05112 during anti-CD3 treatment of the cells did not prevent unresponsiveness . However , the addition of P60568 but not P05112 during P25054 and Lol p I stimulation partially reversed the anergic state . These data demonstrate that , contrary to the mouse , cloned T cells of all three human T helper cell subtypes are anergized by anti-CD3 TCR activation in the absence of costimulatory signals . The fact that human Th2 cells can be anergized may be important for the development of new treatments in Th2-mediated allergic disorders . The expression of P42768 ( WASP ) is dependent on O43516 ( O43516 ) . The P42768 ( WASP ) is a key molecule for transduction of extracellular signals that induce a variety of critical biological events involving actin cytoskeleton rearrangement . Among the cellular partners of WASP , the P42768 -interacting protein ( O43516 ) has been speculated to play a critical role in the pathophysiology of Wiskott-Aldrich syndrome since WASP mutation hot spots map to the O43516 -binding region . The notion that O43516 promotes WASP function , however , conflicts with evidence that O43516 inhibits WASP-mediated actin polymerization and P60568 production and suggests a complex regulation of WASP function by O43516 . Here we show that WASP gene transfer results in high WASP expression only when O43516 is concomitantly expressed in K562 cells . Furthermore , O43516 -knockdown experiments demonstrated that T cells with reduced O43516 expression show a concordant reduction of WASP levels . Mapping studies using O43516 mutants showed that the minimal O43516 region able to rescue WASP expression in O43516 -knockdown cells was the WASP-binding domain . However , expression of such a minimal domain of O43516 failed to rescue WASP-dependent , nuclear factor of activated T-cells-mediated P60568 transcriptional activity . These results demonstrate that expression of O43516 is necessary for functional WASP expression in human cells and provide a new paradigm for understanding the function of these two molecules . P35372 -dependent and independent components in effects of tramadol . DB00193 is thought to induce analgesia via both opioid and non-opioid pathways , although the precise mechanisms remain to be elucidated . In this study , we investigated the roles of the mu-opioid receptor ( MOP ) in analgesic and rewarding effects of tramadol by using MOP knockout ( KO ) mice . DB00193 -induced antinociception , assessed by hot-plate and tail-flick tests , was significantly reduced in heterozygous and homozygous MOP-KO mice when compared with that in wild-type mice . Interestingly , however , tramadol retained its ability to induce significant antinociception in homozygous MOP-KO mice . The tramadol-induced antinociception remaining in homozygous MOP-KO mice was not significantly affected by methysergide , a serotonin receptor antagonist , but was partially blocked by yohimbine , an adrenaline alpha2 receptor antagonist , and both naloxone , a non-selective opioid receptor antagonist , and yohimbine . In addition , antinociceptive effects of an active tramadol metabolite M1 were abolished or remarkably reduced in MOP-KO mice . On the other hand , neither wild-type nor homozygous MOP-KO mice showed significant place preference for tramadol in a conditioned place preference test , although there were slight tendencies toward preference in wild-type mice and avoidance in homozygous MOP-KO mice . These results strongly support the idea suggested in the previous pharmacological studies that MOP and the adrenaline alpha2 receptor mediate most of the analgesic properties of tramadol . Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . Characterization of antihistamines using biphasic cutaneous reaction in BALB/c mice . Effects of 11 histamine H1 receptor antagonists on IgE-mediated biphasic cutaneous reaction in mice were examined . The immediate phase reaction ( IPR ) assessed at 1 hour after antigen application was significantly inhibited by all antihistamines examined . The inhibition of IPR by cetirizine and mequitazine were potent , but those by cyproheptadine and diphenhydramine were weak . The later phase reaction ( LPR ) assessed at 24 hours after antigen application was inhibited by chlorpheniramine , oxatomide , ketotifen , mequitazine , emedastine , terfenadine and azelastine . The inhibition of LPR by emedastine was potent , but those by ketotifen and terfenadine were only partial . DB01084 inhibited both IPR and LPR comparably . Present results indicate that H1 receptor activation is involved in the IPR of the biphasic cutaneous reaction , and that the blockade of H1 receptors at IPR does not contribute to the attenuation of following LPR . P35367 antagonists inhibiting the LPR have a property distinct from H1 receptor antagonism , which may have an additional benefit for the treatment of allergic diseases . Is olomoucine , a weak P24941 inhibitor , able to induce apoptosis in cancer cells ? DB02116 ( OLO ) , a substituted purine analogue , is a much weaker inhibitor of cyclin-dependent kinases than other closely related DB00171 derivatives . It has been recently reported that OLO did not affect the viability of normal human MRC-5 fibroblasts , but it inhibited the proliferation of human HL-60 leukemia cells . Therefore , it was interesting to explore the antiproliferative effect of OLO and to characterize its action on distinct human cancer cells differing in the functional status of the cell cycle . Human HeLa cervical carcinoma and HL-60 leukemia cells were continuously exposed to increasing concentrations of OLO for 24 h and 48 h or alternatively , cells after treatment for 24 h were postincubated in a drug-free medium . Surprisingly , OLO more strongly affected the proliferation of HL-60 cells than that of HeLa cells . Flow cytometric analyses revealed that OLO at higher doses increases the frequency of a hypoploid HL-60 cell population representing cells undergoing apoptosis . These results substantiated the data of the determination of the number of viable cells . Moreover , OLO at higher doses modulates the cell cycle progression of tested cancer cells . Detailed analyses of the DNA concentration in single cells revealed that OLO-mediated reduction of the number of G(1)-phase cells was accompanied by an increase of the frequency of G(2)-phase cells . The kinetics of these changes differed between both tested cancer cell lines , suggesting that some cancer cells exhibit increased susceptibility to OLO action . It remains to clarify whether the strong proapoptotic effect of OLO observed in HL-60 cells depends on their differentiation status . Gene regulation by hypoxia and the neurodevelopmental origin of schizophrenia . Neurodevelopmental changes may underlie the brain dysfunction seen in schizophrenia . While advances have been made in our understanding of the genetics of schizophrenia , little is known about how non-genetic factors interact with genes for schizophrenia . The present analysis of genes potentially associated with schizophrenia is based on the observation that hypoxia prevails in the embryonic and fetal brain , and that interactions between neuronal genes , molecular regulators of hypoxia , such as hypoxia-inducible factor 1 ( Q9BYW2 ) , and intrinsic hypoxia occur in the developing brain and may create the conditions for complex changes in neurodevelopment . Consequently , we searched the literature for currently hypothesized candidate genes for susceptibility to schizophrenia that may be subject to ischemia-hypoxia regulation and/or associated with vascular expression . Genes were considered when at least two independent reports of a significant association with schizophrenia had appeared in the literature . The analysis showed that more than 50 % of these genes , particularly P31749 , P23560 , O75052 , P32238 , P36544 , P21554 , P21964 , DNTBP1 , Q99259 , Q14832 , P22301 , MLC1 , Q99466 , Q02297 , P43354 / P43354 , O43272 , P78509 , P49798 , Q9NQC3 / Q9NQC3 and P01375 , are subject to regulation by hypoxia and/or are expressed in the vasculature . Future studies of genes proposed as candidates for susceptibility to schizophrenia should include their possible regulation by physiological or pathological hypoxia during development as well as their potential role in cerebral vascular function . Pelvic fracture mechanism of injury in vehicular trauma patients . To investigate the correlation between motor vehicle crash mechanisms and pelvic injury in front-seat occupants , we retrospectively reviewed the clinical records of , and had complete crash reconstructions performed for , 145 vehicular trauma patients with injury Severity Scores greater than 16 admitted to a level I trauma center . After excluding rear-seat and ejected occupants , 44 of the remaining 115 patients had pelvic injuries . We excluded acetabular fractures and classified the remaining 26 pelvic ring fractures by the system of Young and Burgess : 20 lateral compression ( LC ) fractures , five anteroposterior compression ( P25054 ) fractures , and one combined mechanical injury ( CMI ) fracture . Eighteen pelvic fractures were managed conservatively ; eight required surgical intervention and four of those eight required emergent application of an external fixator for unresponsive hypotension . Trained investigation teams conducted the crash reconstructions , evaluating crash sites and vehicles with direct measurements of more than 500 variables . Calculations from these data , e.g. , direction of impact and change in velocity at impact ( delta V ) , were made with the Q7L266 III computer program and statistical analyses were performed using Chi-square and t tests . This information was then merged with the orthopedic evaluations. ( ABSTRACT TRUNCATED AT 250 WORDS )

MH_train_1395

MH_train_1395

MH_train_1395

interacts_with DB01114?

multiple_choice

Increased P05231 levels in pituitary-deficient patients are independently related to their carotid intima-media thickness . OBJECTIVE : Increased cardiovascular morbidity and mortality has been observed in patients with pituitary deficiency and untreated growth hormone deficiency ( GHD ) . We investigated peripheral inflammatory and fibrinolytic markers and their associations with arterial intima-media thickness ( IMT ) in GHD . DESIGN : Cross-sectional study . PATIENTS : Thirty-four patients with GHD , but without cardiovascular disease , were compared to healthy controls matched for age , sex , body mass index ( BMI ) and smoking habits . MEASUREMENTS : IMT of the common carotid artery , P02741 ( CRP ) , interleukin-6 ( P05231 ) , fibrinogen , plasminogen activator inhibitor-1 ( P05121 ) activity and tissue plasminogen activator antigen ( tPA-ag ) were measured . RESULTS : Median P05231 concentrations were increased by 208 % and 248 % in GHD patients compared to BMI-matched and nonobese controls , respectively . Median CRP and tPA-ag levels were increased by 237 % and 167 % in patients compared to nonobese controls , but not significantly different compared to BMI-matched controls . Plasma levels of fibrinogen and P05121 activity did not differ between groups . Age , low-density lipoprotein ( LDL ) cholesterol , tPA-ag and P05231 were positively correlated , and P05019 was negatively correlated to IMT in the patient group , but only age and P05231 were independently related to IMT . CONCLUSIONS : P05231 concentrations were increased in GHD patients compared to controls and independently related to IMT in patients . This finding may help to explain the variance in IMT and the increased vascular morbidity and mortality in hypopituitary patients with GHD . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . DB02426 effects on brown-fat mitochondria imply that the adenine nucleotide translocator isoforms P12235 and P05141 may be responsible for basal and fatty-acid-induced uncoupling respectively . In brown-fat mitochondria , fatty acids induce thermogenic uncoupling through activation of P25874 ( uncoupling protein 1 ) . However , even in brown-fat mitochondria from P25874 -/- mice , fatty-acid-induced uncoupling exists . In the present investigation , we used the inhibitor CAtr ( carboxyatractyloside ) to examine the involvement of the ANT ( adenine nucleotide translocator ) in the mediation of this P25874 -independent fatty-acid-induced uncoupling in brown-fat mitochondria . We found that the contribution of ANT to fatty-acid-induced uncoupling in P25874 -/- brown-fat mitochondria was minimal ( whereas it was responsible for nearly half the fatty-acid-induced uncoupling in liver mitochondria ) . As compared with liver mitochondria , brown-fat mitochondria exhibit a relatively high ( P25874 -independent ) basal respiration ( ' proton leak ' ) . Unexpectedly , a large fraction of this high basal respiration was sensitive to CAtr , whereas in liver mitochondria , basal respiration was CAtr-insensitive . Total ANT protein levels were similar in brown-fat mitochondria from wild-type mice and in liver mitochondria , but the level was increased in brown-fat mitochondria from P25874 -/- mice . However , in liver , only Ant2 mRNA was found , whereas in brown adipose tissue , Ant1 and Ant2 mRNA levels were equal . The data are therefore compatible with a tentative model in which the P05141 isoform mediates fatty-acid-induced uncoupling , whereas the P12235 isoform may mediate a significant part of the high basal proton leak in brown-fat mitochondria . Further characterization of a somatic cell hybrid panel : ten new assignments to the bovine genome . Thirty-six partially characterized hamster-bovine hybrid cell lines were used for the determination of synteny groups . Sixteen additional reference loci , selected for their coverage of the bovine genome , were analysed on these hybrid cells . This increases to 25 the number of synteny groups detected . This panel was then used to make synteny assignments for 10 additional loci , eight by Southern blotting ( P02452 , P08123 , FAS , P07858 , P07711 , P07510 , P07686 and P08908 ) and two by polymerase chain reaction ( PCR ) amplification ( P35367 and ETH1112 ) . These loci were assigned to international synteny groups U12 ( P35367 ) , U13 ( P08123 ) , U17 ( P07510 ) , U21 ( P02452 , FAS ) , U29 ( ETH1112 ) , to chromosome 20 ( U14 or U25 ) for P07686 and P08908 , and to the same local synteny group ( A ) , which is probably U18 , for P07858 and P07711 . For three loci already mapped in humans ( P02452 , P08123 and P07510 ) , the present results are in accordance with the predictions based on comparative mapping between the human and bovine species . Effects of lesopitron on the central nervous system arising from its interaction with P08908 receptors . DB04970 acts as a ligand for central serotonin P08908 receptors . Ki obtained from [3H]8-OH-DPAT competition studies was 104.8 +/- 10.6 nmol/l . As lesopitron did not affect the binding of [3H]paroxetine , involvement of the serotonin reuptake system in the effects of lesopitron is rejected . DB04970 inhibits haloperidol-induced catalepsy that is the consequence of its action on P08908 autoreceptors . The ability of lesopitron to induce 5-HT syndrome reflects post-synaptic P08908 receptor activation and the reversion of 8-OHDPAT-induced 5-HT syndrome by lesopitron suggests a partial agonist effect on this receptor-type . DB04970 induced a hypothermic effect due to the enhanced activation of post-synaptic P08908 receptors . The agonist effect of lesopitron on P08908 receptors and its marked hypothermic effect is an added value for this drug and a stimulus to the study of its possible neuroprotective action . DB11320 modulates multiple functional activities of monocyte-derived dendritic cell subsets via histamine receptor 2 . Expression of CD1a proteins in human monocyte-derived dendritic cells ( DCs ) specifies functionally distinct subsets with different inflammatory properties . DB11320 is recognized as an inflammatory mediator released by various cell types including DCs . The diverse biological effects of histamine are mediated by G-protein-coupled histamine receptors ( HRs ) , which are able to modulate the functional activities of DC subsets . The goal of the present study was to compare the expression and activity of HRs in the CD1a(-) and CD1a(+) monocyte-derived DC subsets and to test the effects of histamine on the differentiation , activation and functional activities of these subsets . We show that P25021 is present at high levels in both DC subsets , whereas P35367 and Q9H3N8 are expressed in a subset-specific manner . DB11320 shifts DC differentiation to the development of CD1a(-) DCs and modulates DC activation through its inhibitory effect on CD1a(+) DC differentiation . DB11320 -induced reduction of CD1a(+) DCs is associated with increased secretion of P05231 and P22301 , up-regulation of a typical combination of chemokines , expression C5aR1 by the CD1a(-) DC subset and enhanced migration of both activated DC subsets supported by the production of P14780 and P39900 enzymes . All these effects were shown to be mediated in a P25021 -specific manner as revealed by the specific antagonist of the receptor . As P25021 is expressed at high levels in both DC subsets , we propose that it may dominate the regulation of multiple DC functions . In contrast , P35367 and Q9H3N8 with opposing subset-related expression may have a regulatory or fine-tuning role in histamine-induced functional activities . Clinical and genetic factors associated with nausea and vomiting in cancer patients receiving opioids . BACKGROUND : This study investigates whether demographical , disease-related and genetic factors contribute to inter-individual differences in nausea and vomiting among patients receiving opioids for cancer pain . METHODS : Cancer patients receiving opioids were included from 17 centres in 11 European countries . Intensities of nausea and vomiting were reported by 1579 patients on four-point categorical scales . In stratified regression models including demographical and disease-related factors as covariates , 96 single nucleotide polymorphisms ( SNPs ) in 16 candidate genes related to opioid- or nausea/vomiting signalling pathways ( P08183 , P35372 , P41145 , P32121 , P42226 , P21964 , P20309 , P08912 , P35367 , P14416 , P35462 , P25103 , P46098 , O95264 , Q8WXA8 , P21554 ) were analysed for association with nausea and vomiting . FINDINGS : Age , body mass index , Karnofsky Performance Status , gender , use of antiemetics , type of opioid , type of cancer and eight SNPs were associated with the inter-individual differences in nausea and vomiting among cancer patients treated with opioids ( p < 0.01 ) . The SNPs were rs1176744 , rs3782025 and rs1672717 in O95264 ; rs165722 , rs4680 and rs4633 in P21964 ; rs10802789 and rs685550 in P20309 . Only the SNP rs1672717 in O95264 passed the Benjamini-Hochberg criterion for a 10 % false discovery rate . INTERPRETATION : Clinical characteristics and SNPs within the O95264 , P21964 and P20309 genes may be associated with the variability in nausea and vomiting among cancer patients receiving opioids . This knowledge may help to identify patients at particular risk for nausea and vomiting during treatment with opioids for cancer pain . A fully human recombinant IgG-like bispecific antibody to both the epidermal growth factor receptor and the insulin-like growth factor receptor for enhanced antitumor activity . Both the epidermal growth factor receptor ( P00533 ) and the insulin-like growth factor receptor ( IGFR ) have been implicated in the tumorigenesis of a variety of cancers . Here we propose that simultaneous targeting of both receptors with a bispecific antibody would lead to enhanced antitumor activity . To this end , we produced a recombinant human IgG-like bispecific antibody , a Di-diabody , using the variable regions from two antagonistic antibodies : DB05774 to P00533 and DB05759 to IGFR . The Di-diabody binds to both P00533 and IGFR and effectively blocked both P01133 - and IGF-stimulated receptor activation and tumor cell proliferation . The Di-diabody also inherited the biological properties from both of its parent antibodies ; it triggers rapid and significant IGFR internalization and degradation and mediates effective antibody-dependent cellular cytotoxicity in a variety of tumor cells . Finally , the Di-diabody strongly inhibited the growth of two different human tumor xenografts in vivo . Our results underscore the benefits of simultaneous targeting of two tumor targets with bispecific antibodies . Expression of the human concentrative nucleotide transporter 1 ( O00337 ) gene correlates with clinical response in patients affected by Waldenström 's Macroglobulinemia ( WM ) and small lymphocytic lymphoma ( SLL ) undergoing a combination treatment with 2-chloro-2'-deoxyadenosine ( DB00242 ) and DB00073 . PURPOSE : Resistance to nucleoside analogues agents is likely to be multifactorial and could involve a number of mechanisms affecting drug penetration , metabolism and targeting . In vitro studies of resistant human cell lines have confirmed that human concentrative nucleoside transporter 1 ( O00337 ) -deficient cells display resistance . EXPERIMENTAL DESIGN : We applied real-time PCR method to assess the mRNA expression of equilibrative and concentrative nucleoside transporter ( hENT1 , O00337 ) , deoxycytidine and deoxyguanosine kinase ( P27707 , Q16854 ) , 5'-nucleotidase ( 5'-NT ) , ribonucleotide reductase catalytic and regulatory ( P23921 , P31350 ) subunits in bone marrow cells from 32 patients with Waldenström 's Macroglobulinemia ( WM ) and small lymphocytic lymphoma ( SLL ) who received 2CdA-based chemotherapy . Responses to chemotherapy , were then correlated to the expression of these markers . RESULTS : All 32 patients enrolled expressed lower levels of O00337 as compared to healthy donors . In univariate analysis , lower expression level of O00337 ( p=0.0021 ) and P31350 ( p=0.02 ) correlated with response to chemotherapy . In particular , patients with low levels of O00337 achieved inferior clinical response . No significant correlation between these genes expression and age , stage of disease was found . This study suggests that nucleotidase expression levels can be used to identify subgroups of WM and SLL patients who will likely respond differently to a 2CdA-based therapy . Evaluation of platelet activation , coagulation , and fibrinolytic activation in patients with symptomatic lacunar stroke . BACKGROUND : It is unclear whether hemostasis plays a role in the pathogenesis of ischemic stroke subtypes . OBJECTIVE : We aimed to investigate the possible relationship between different hemostatic markers and lacunar stroke . RESULTS : The study consisted of 30 patients with symptomatic lacunar stroke and 30 healthy age-matched healthy individuals . We analyzed the values of " Mean Platelet Volume , " D-dimer , " soluble p-selectin , " " P00747 Activator Inhibitor Type-1 " ( P05121 ) , " Thrombin-Activatable DB06692 " ( Q96IY4 ) , and " Platelet Factor 4 " ( P02776 ) in patients with lacunar infarct and compared these values to those of control individuals . There were significant differences for D-dimer , mean platelet volume , thrombin-activatable fibrinolysis inhibitor , and platelet factor 4 values in symptomatic lacunar stroke group compared with the control group ( P < 0.01 ) . CONCLUSIONS : Different hemostatic factors may play a role in the pathogenesis of lacunar stroke . Evaluating the role of hemostatic factors on different types of strokes may help us identify new therapeutic strategies and different prognostic stratifications for ischemic stroke . Role of nitrative and oxidative DNA damage in inflammation-related carcinogenesis . Chronic inflammation induced by biological , chemical , and physical factors has been found to be associated with the increased risk of cancer in various organs . We revealed that infectious agents including liver fluke , Helicobacter pylori , and human papilloma virus and noninfectious agents such as asbestos fiber induced P35228 -dependent formation of 8-nitroguanine and 8-oxo-7 , 8-dihydro-2'-deoxyguanosine ( 8-oxodG ) in cancer tissues and precancerous regions . Our results with the colocalization of phosphorylated Q13315 and γ- P16104 with 8-oxodG and 8-nitroguanine in inflammation-related cancer tissues suggest that DNA base damage leads to double-stranded breaks . It is interesting from the aspect of genetic instability . We also demonstrated P05231 -modulated P35228 expression via P40763 and P00533 in Epstein-Barr-virus-associated nasopharyngeal carcinoma and found promoter hypermethylation in several tumor suppressor genes . Such epigenetic alteration may occur by controlling the DNA methylation through P05231 -mediated JAK/ P40763 pathways . Collectively , 8-nitroguanine would be a useful biomarker for predicting the risk of inflammation-related cancers . Assessment of partially deoxygenated deoxynojirimycin derivatives as glucosylceramide synthase inhibitors . Q16739 ( Q16739 ) 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 ) 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 -hydroxyl of the deoxynojirimycin core is important for Q16739 inhibition . Here we show that P13671 -OH appears of less important , which may set guidelines for the development of Q16739 inhibitors that have less affinity ( in comparison with DB00419 ) for other glycoprocessing enzymes , in particular those hydrolases that act on glucosylceramide . Cytokine regulation of intercellular adhesion molecule-1 ( P05362 ) expression on human glioblastoma cells . Intercellular adhesion molecule-1 ( P05362 ) has recently been identified as one of the ligands for lymphocyte function-associated antigen-1 ( LFA-1 ) . Immunohistochemical staining of frozen tissue sections using the P05362 antibody P23921 /1 demonstrated significant levels of P05362 expression on human glioblastoma cells and on intratumoural vascular endothelial cells . P05362 was weakly expressed or absent from low grade gliomas and absent from normal and fetal brain . P05362 expression was similar to that of MHC class II . HLA-DR antigens . Glioblastoma cell lines constitutively expressed P05362 to a minimal or moderate extent . Surface antigen expression of P05362 and P05362 -specific mRNA could be significantly increased by incubating glioblastoma cells with interleukin-1 beta ( P01584 ) , tumour necrosis factor-alpha ( P01375 ) , and interferon-gamma ( P01579 ) . P60568 , P05112 , P05231 and transforming growth factor beta 2 ( TGF-beta 2 ) had no significant effect on surface antigen expression . Significant enhancement of P05362 expression was obtained using P01375 and P01584 at 1-10 U/ml and at 500 U/ml of P01579 . Induction of P05362 specific mRNA was observed 4 h after cytokine treatment and decreased by 24 h . Surface antigen expression of P05362 increased for up to 48 h after treatment . Expression of membrane complement regulators , P15529 , P08174 and P13987 , in mesothelial cells of patients on peritoneal dialysis therapy . We investigated the expression of membrane complement regulators ( CRegs ) , P15529 , P08174 and P13987 in human mesothelial cells , and correlated with clinical background and level of complement ( C ) activation products in peritoneal dialysis ( PD ) fluids ( PDF ) to clarify influence of the C activation system in PD patients . Expression of CRegs was assessed on primary cultures of mesothelial cells ( HPMC ) harvested from PD fluid of 31 PD patients . Because expression of P08174 but not P15529 and P13987 in mesothelial cells was significantly correlated to value of dialysate-to-plasma creatinine concentration ratio ( D/P Cre ) ( p < 0.005 ) as an indicator of peritoneal function , we focused on analysis of P08174 expression of HPMCs in comparison with levels of C activation products in the PDF of the PD patients , and their background factors . When comparing expression of the CRegs between systemic neutrophils and HPMC , no correlation was observed , supporting that change of CRegs ' expression in HPMC was independently occurring in the peritoneum . Expression of P08174 protein in HPMC was closely correlated with expression at the mRNA level ( p < 0.0001 ) and was inversely correlated with levels of sC5b-9 ( p < 0.05 ) , but not P01024 , C4 , P05231 and Q8WXI7 in the PDF . Complications of diabetes , usage of icodextrin and residual renal function were not correlated with change of P08174 expression in HPMCs . Our data show that the process of PD therapy modifies expression of P08174 on peritoneal mesothelium and triggers local C activation . These findings support efforts to modify PD therapy to limit effects on activation and regulation of the C system . P35367 occupancy in human brains after single oral doses of histamine H1 antagonists measured by positron emission tomography . 1. P35367 occupancy in the human brain was measured in 20 healthy young men by positron emission tomography ( PET ) using [ 11C ] -doxepin . 2 . (+)- DB01114 , a selective and classical antihistamine , occupied 76.8 +/- 4.2 % of the averaged values of available histamine H1 receptors in the frontal cortex after its administration in a single oral dose of 2 mg . Intravenous administration of 5 mg (+)-chlorpheniramine almost completely abolished the binding of [ 11C ] -doxepin to H1 receptors ( H1 receptor occupancy : 98.2 +/- 1.2 % ) . 3 . Terfenadine , a nonsedative antihistamine , occupied 17.2 +/- 14.2 % of the available H1 receptors in the human frontal cortex after its administration in a single oral dose of 60 mg . 4 . There was no correlation between H1 receptor occupancy by terfenadine and the plasma concentration of the active acid metabolite of terfenadine in each subject . 5 . PET data on human brain were essentially compatible with those on H1 receptor occupancy in guinea-pig brain determined by in vivo binding techniques , although for the same H1 receptor occupancy the dose was less in human subjects than in guinea-pigs . 6 . The PET studies demonstrated the usefulness of measuring H1 receptor occupancy with classical and second-generation antihistamines in human brain to estimate their unwanted side effects such as sedation and drowsiness quantitatively . DB00125 -vasopressin directly promotes a thermogenic and pro-inflammatory adipokine expression profile in brown adipocytes . DB00125 -vasopressin ( AVP ) - via activation of the hypothalamic-pituitary-adrenal ( Q9Y251 ) axis - may play a role in the regulation of energy homeostasis and related cardiovascular complications . Brown adipose tissue ( Q14032 ) - via dissipation of energy in the form of heat - contributes to whole body energy balance . Q14032 expresses vasopressin receptors . We investigated direct effects of AVP on brown adipose endocrine and metabolic functions . P25874 -1 protein expression in differentiated brown adipocytes was induced after acute exposure of adipocytes to AVP . This effect was time-dependent with a maximum increase after 8h . AVP also induced a time- and dose-dependent increase in p38 Q96HU1 kinase phosphorylation . Pharmacological inhibition of p38 Q96HU1 kinase with SB 202190 abolished the induction of P25874 -1 protein expression . Furthermore , while acute AVP treatment enhanced mRNA expression of P13500 and P05231 , adiponectin mRNA expression was reduced . Yet , on the level of intracellular glucose uptake , there was no AVP-induced change of adipose insulin-induced glucose uptake . Finally , there was no difference in lipid accumulation between control and AVP-treated cells . Taken together , our data demonstrate direct effects of AVP on thermogenic , inflammatory , and glucoregulatory gene expression in brown adipocytes , thus expanding the hitherto known spectrum of this neuropeptides 's biological effects and suggesting a direct adipotropic role as a stress-promoting factor . Improved soluble expression of a single-chain antibody fragment in E. coli for targeting Q8WXI7 in epithelial ovarian cancer . Production of antibody fragments in heterologous hosts such as Escherichiacoli provides a unique and cost-effective method to develop engineered vectors for tumor targeting . A single-chain Fragment variable ( scFv ) of the murine monoclonal antibody MAb- DB04964 targeting Q8WXI7 in epithelial ovarian cancer was previously developed , expressed , purified and proposed as a functional targeting entity for biomedical applications . However , the yields from its soluble expression in heterologous systems were very low for any practical use in preclinical translational research ; leave alone the defeated objective of convenient and cost-effective production . In the present work , the anti- Q8WXI7 scFv gene was re-organized and sub-cloned into pET-22b(+) vector to be in frame with the pelB leader peptide for periplasmic localization and C-terminal hexa-histidine tag to facilitate downstream purification . Six variants of the scFv were constructed to investigate the impact of variable domain orientations , inter-domain peptide linker sequences and codon optimization on the soluble expression of the scFv using Rosetta 2(DE3) as the E. coli host supplemented with tRNAs for rare codons . Expression in shake flask cultures under the control of an inducible T7 promoter and subsequent purification by cobalt based immobilized metal affinity chromatography yielded differential amounts of high purity scFv for all constructs . Here , we report up to 14-fold increase in the soluble expression of the scFv primarily as a result of codon optimization with minor effects from inter-domain peptide linkers and variable domain orientation in the anti- Q8WXI7 scFv molecule . All the scFv constructs expressed and purified were found to be immunoreactive for in vitro targeting of Q8WXI7 antigen . P35372 phosphorylation , desensitization , and ligand efficacy . Mu opioid receptors are subject to phosphorylation and desensitization through actions of at least two distinct biochemical pathways : agonist-dependent mu receptor phosphorylation and desensitization induced by a biochemically distinct second pathway dependent on protein kinase C activation ( 1 ) . To better understand the nature of the agonist-induced mu receptor phosphorylation events , we have investigated the effects of a variety of opiate ligands of varying potencies and intrinsic activities on mu receptor phosphorylation and desensitization . Exposure to the potent full agonists sufentanil , dihydroetorphine , etorphine , etonitazine , and [ D-Ala2 , MePhe4 , Glyol5 ] enkephalin ( DAMGO ) led to strong receptor phosphorylation , while methadone , l-alpha-acetylmethadone ( DB01227 ) , morphine , meperidine , DADL , beta-endorphin(1-31) , enkephalins , and dynorphin A(1-17) produced intermediate effects . The partial agonist buprenorphine minimally enhanced receptor phosphorylation while antagonists failed to alter phosphorylation . DB00921 and full antagonists each antagonized the enhanced mu receptor phosphorylation induced by morphine or DAMGO . The rank order of opiate ligand efficacies in producing mu receptor-mediated functional desensitization generally paralleled their rank order of efficacies in producing receptor phosphorylation . Interestingly , the desensitization and phosphorylation mediated by methadone and DB01227 were disproportionate to their efficacies in two distinct test systems . This generally good fit between the efficacies of opiates in mu receptor activation , phosphorylation , and desensitization supports the idea that activated receptor/agonist/G-protein complexes and/or receptor conformational changes induced by agonists are required for agonist-induced mu receptor phosphorylation . Data for methadone and DB01227 suggest possible contribution from their enhanced desensitizing abilities to their therapeutic efficacies . Endogenous expression of histamine H1 receptors functionally coupled to phosphoinositide hydrolysis in P13671 glioma cells : regulation by cyclic AMP . 1. The effects of histamine receptor agonists and antagonists on phospholipid hydrolysis in rat-derived P13671 glioma cells have been investigated . 2 . P35367 -stimulation caused a concentration-dependent increase in the accumulation of total [ 3H ] -inositol phosphates in cells prelabelled with [ 3H ] -myo-inositol . The rank order of agonist potencies was histamine ( EC50 = 24 microM ) > N alpha-methylhistamine ( EC50 = 31 microM ) > 2-thiazolylethylamine ( EC50 = 91 microM ) . 3 . The response to 0.1 mM histamine was antagonized in a concentration-dependent manner by the H1-antagonists , mepyramine ( apparent Kd = 1 nM ) and (+)-chlorpheniramine ( apparent Kd = 4 nM ) . In addition , (-)-chlorpheniramine was more than two orders of magnitude less potent than its (+)-stereoisomer . 4 . Elevation of intracellular cyclic AMP accumulation with forskolin ( 10 microM , EC50 = 0.3 microM ) , isoprenaline ( 1 microM , EC50 = 4 nM ) or rolipram ( 0.5 mM ) , significantly reduced the histamine-mediated ( 0.1 mM ) inositol phosphate response by 37 % , 43 % and 26 % respectively . In contrast , 1,9-dideoxyforskolin did not increase cyclic AMP accumulation and had no effect on the phosphoinositide response to histamine . 5 . These data indicate the presence of functionally coupled , endogenous histamine H1 receptors in P13671 glioma cells . Furthermore , the results also indicate that H1 receptor-mediated phospholipid hydrolysis is inhibited by the elevation of cyclic AMP levels in these cells . Matrix proteinase inhibition by DB05387 , a multifunctional antiangiogenic compound . BACKGROUND : Matrix metalloproteinases ( MMPs ) play an important role in tissue remodeling under normal physiological and pathological conditions and are thus attractive targets for both diagnostic and therapeutic purposes . Here , we examined the effect of DB05387 , an orally bioavailable standardized extract made of cartilage that shows significant antiangiogenic and antimetastatic properties in vivo , on the activity of various members of the MMP family . MATERIALS AND METHODS : The effect of DB05387 on the activity of MMPs was assessed by fluorimetric assays and by substrate gel zymography . RESULTS : DB05387 markedly inhibits the gelatinolytic activity of P08253 and to a lesser extent those of P03956 , P09237 , P14780 and P45452 . DB05387 also inhibited the elastinolytic activities of P08253 and P14780 as well as P39900 ( metalloelastase ) , porcine pancreatic elastase ( PPE ) , and human leukocyte elastase ( P08246 ) . Western blot analysis revealed the presence within DB05387 of immunoreactive P01033 -like proteins , suggesting that these proteins may be at least partly responsible for the observed MMP inhibition . CONCLUSIONS : Taken together , these results demonstrate that DB05387 contains P01033 -like proteins that could be responsible for the specific inhibition of MMPs . Given the recent studies suggesting the presence within this compound of specific inhibitor(s) of endothelial cell proliferation , DB05387 appears as a pleotropic agent able to interfere with several biochemical steps leading to angiogenesis and to other physiopathological conditions . Since DB05387 is currently under Phase III clinical investigations , these findings are also of considerable importance for our understanding of its anticancer properties .

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MH_train_1396

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interacts_with DB00668?

multiple_choice

DB00945 resistance : clinical significance and genetic polymorphism . OBJECTIVE : To determine the prevalence , clinical implications and underlying mechanism of aspirin resistance in Chinese patients . METHODS : Platelet aggregation was determined by light transmission aggregometry ( P01374 ) using four different inducers . Patients were divided into aspirin-resistant ( AR ) , aspirin semi responder ( ASR ) and aspirin-sensitive ( AS ) groups , according to their P01374 results . DB00945 resistance was assessed by thrombo elastography ( TEG , with arachidonic acid [ AA ] or adenosine diphosphate as inducers ) , serum/urinary 11-dehydrothromboxane B2 ( P28845 -TXB2 ) assay , platelet function analyser-100 assay and P16109 assay . Polymorphisms in the prostaglandin endoperoxide synthase 1 ( P23219 ) gene ( A842G , C50T , C22T , G128A , C644A and C714A ) , the P35354 gene ( G765C ) and the integrin β3 ( P05106 ) gene ( C196T ) were examined . RESULTS : The study included 360 aspirin-treated patients and 314 healthy controls . AS patients had significantly lower levels of P28845 -TXB2 than AR and ASR patients , and significantly lower levels of P16109 than AR patients . TEG-AA was more sensitive , specific and consistent than P16109 in detecting aspirin resistance . The frequency of the P35354 G765C mutation was significantly higher in the AR/ASR groups versus the AS group . CONCLUSIONS : TEG-AA was more sensitive , specific and consistent than the P16109 assay for detecting aspirin resistance , and the P35354 G765C mutation may be related to aspirin resistance . 5-Hydroxytryptamine stimulates cyclic AMP formation in the tunica muscularis mucosae of the rat oesophagus via Q13639 receptors . The nature of Q13639 receptor coupling in the tunica muscularis mucosae of the rat oesophagus has been studied . 5-HT and renzapride stimulated cyclic AMP formation concentration dependently , with -log EC50 values of 7.1 and 6.8 , respectively . DB04917 , relative to 5-HT , acted as a partial agonist . Tropisetron ( ICS 205 930 ) and a novel Q13639 antagonist , SDZ 205 557 , inhibited 5-HT-induced cyclic AMP production competitively , with pA2 estimates of 6.7 and 7.7 , respectively . These data are consistent with the hypothesis that Q13639 receptors mediate relaxation of the smooth muscle cells of the tunica muscularis mucosae of rat oesophagus via activation of adenylyl cyclase . DB00877 unbalances the polarization of human macrophages to M1 . Plasticity is a hallmark of macrophages , and in response to environmental signals these cells undergo different forms of polarized activation , the extremes of which are called classic ( M1 ) and alternative ( M2 ) . DB00877 ( Q96PN7 ) is crucial for survival and functions of myeloid phagocytes , but its effects on macrophage polarization are not yet studied . To address this issue , human macrophages obtained from six normal blood donors were polarized to M1 or M2 in vitro by lipopolysaccharide plus interferon-γ or interleukin-4 ( P05112 ) , respectively . The presence of Q96PN7 ( 10 ng/ml ) induced macrophage apoptosis in M2 but not in M1 . Beyond the impact on survival in M2 , Q96PN7 reduced P61073 , CD206 and Q9NNX6 expression and stem cell growth factor-β , P55774 and Q99616 release . In contrast , in M1 Q96PN7 increased P42081 and P32248 expression and P05231 , tumour necrosis factor-α and IL-1β release but reduced CD206 and Q9NNX6 expression and P22301 , vascular endothelial growth factor and P55774 release . In view of the in vitro data , we examined the in vivo effect of Q96PN7 monotherapy ( 0·1 mg/kg/day ) in 12 patients who were treated for at least 1 month before islet transplant . Cytokine release by O00206 -stimulated peripheral blood mononuclear cells showed a clear shift to an M1-like profile . Moreover , macrophage polarization 21 days after treatment showed a significant quantitative shift to M1 . These results suggest a role of mammalian target of rapamycin ( P42345 ) into the molecular mechanisms of macrophage polarization and propose new therapeutic strategies for human M2-related diseases through P42345 inhibitor treatment . Sensitivity of receptors for IgA on T560 , a murine B lymphoma , to phorbol myristate acetate and to phosphatidylinositol-specific phospholipase C . A P07902 -derived B lymphoma , T560 , that bears IgAR is described . T560 is IgG2a kappa + , Ia+ , B220+ , J11d+ , Thy-1- , CD3- , P01730 - , P06127 - , Mac 1- , Mac 2- , nonspecific esterase negative and binds bromelain-treated mouse RBC but not SRBC or ORBC . It presents antigen , secretes IL-1 , P05112 and P05231 but not P60568 , P05113 or TGF beta and appears to be related to the Lyt 1+( P06127 ) lineage of B cells though it lacks Lyt 1 . T560 bears IgAR that , on the cell surface , are completely cross-inhibited by low concentrations of IgM and by high concentrations of IgG2a and IgG2b . They do not appear to represent a cell-surface form of galactosyl transferase . They are inducible by high concentrations of IgA , sensitive to trypsin and insensitive to neuraminidase . They are down-regulated by activation of PKC with PMA , but their recovery is not inhibited by cycloheximide , indicating that they are not degraded or shed . They may either lose their affinity for IgA or be internalized without degradation . Seventy percent of IgA receptor activity is lost when T560 is treated with PI- P98160 ; part of this loss of activity is due to activation of PKC and is inhibited by staurosporine , but approximately 30 % of it is not protected by staurosporine indicating that some , or all , of the IgA receptor of T560 is connected to the cell membrane via a P06744 linker . The T560 IgA receptor could be related to the poly-Ig or M cell receptor. ( ABSTRACT TRUNCATED AT 250 WORDS ) Anti-inflammatory secondary metabolites from the leaves of Rosa laevigata . Bioassay-guided fractionation of a n-BuOH-soluble extract of the leaves of Rosa laevigata led to the isolation of three new 19-oxo-18,19-seco-ur-sane-type triterpenoids , laevigins A-C ( 1-3 ) , a new oleanane-type triterpenoid saponin , laevigin D ( 4 ) , a new geranylmethylbenzoate , 5- [ ( 2″E,6″S ) -6″,7″-dihydroxy-3″,7″-dimethyl-2″-octen-1″-yl ] -2-(β-D-glucopyranosyloxy)-methyl benzoate ( 5 ) , together with 9 known compounds ( 6-14 ) . Their structures were elucidated by spectroscopic and chemical methods . Compounds 4 , 9 , 11 , and 12 significantly suppressed the LPS-stimulated NF-κB transcriptional activity and the release of TNFα , IL-1β , P05231 , and P22301 in mouse RAW 264.7 macrophages . The compound 12 exhibited moderate inhibition on NF-κB transcriptional activity with an IC50 value of 23.21 μM . The IC50 values of compound 12 were measured as 14.32 , 8.53 , 8.04 , and 10.38 μM for the inhibitory activity on TNFα-release , IL-1β-release , P05231 -release , and P22301 -release , respectively . DB00174 synthetase : regulation by cell stress and involvement in tumor biology . DB00174 synthetase ( P08243 ) catalyzes the conversion of aspartate and glutamine to asparagine and glutamate in an DB00171 -dependent reaction . The enzyme is ubiquitous in its organ distribution in mammals , but basal expression is relatively low in tissues other than the exocrine pancreas . Human P08243 activity is highly regulated in response to cell stress , primarily by increased transcription from a single gene located on chromosome 7 . Among the genomic elements that control P08243 transcription is the C/EBP- P39905 response element ( CARE ) within the promoter . Protein limitation or an imbalanced dietary amino acid composition activate the P08243 gene through the amino acid response ( AAR ) , a process that is replicated in cell culture through limitation for any single essential amino acid . Endoplasmic reticulum stress also increases P08243 transcription through the Q9NZJ5 -eIF2- P18848 arm of the unfolded protein response ( UPR ) . Both the AAR and UPR lead to increased synthesis of P18848 , which binds to the CARE and induces P08243 transcription . Elevated expression of P08243 protein is associated with resistance to asparaginase therapy in childhood acute lymphoblastic leukemia and may be a predictive factor in drug sensitivity for certain solid tumors as well . Activation of the Q9P2K8 -eIF2- P18848 signaling pathway , leading to increased P08243 expression appears to be a component of solid tumor adaptation to nutrient deprivation and/or hypoxia . Identifying the roles of P08243 in fetal development , tissue differentiation , and tumor growth may reveal that P08243 function extends beyond asparagine biosynthesis . Inhibition of O00206 with eritoran attenuates myocardial ischemia-reperfusion injury . BACKGROUND : We previously reported that the functional mutation of O00206 ( O00206 ) in C3H/HeJ mice subjected to myocardial ischemia-reperfusion ( MI/R ) injury resulted in an attenuation of myocardial infarction size . To investigate the ligand-activating O00206 during MI/R injury , we evaluated the effect of eritoran , a specific O00206 antagonist , on MI/R injury , with the goal of defining better therapeutic options for MI/R injury . METHODS AND RESULTS : C57BL/6 mice received eritoran ( 5 mg/kg ) intravenously 10 minutes before 30 minutes of in situ of transient occlusion of the left anterior descending artery , followed by 120 minutes of reperfusion . Infarct size was measured using triphenyltetrazoliumchloride staining . A c-Jun NH(2)-terminal kinase ( JNK ) activation was determined by Western blotting , nuclear factor ( NF ) -kappaB activity was detected by gel-shift assay , and cytokine expression was measured by ribonuclease protection assay . Mice treated with eritoran developed significantly smaller infarcts when compared with mice treated with vehicle alone ( 21.0+/-6.4 % versus 30.9+/-13.9 % ; P=0.041 ) . DB04933 pretreatment resulted in a reduction in JNK phosphorylation ( eritoran versus vehicle : 3.98+/-0.81 versus 7.01+/-2.21-fold increase ; P=0.020 ) , less nuclear NF-kappaB translocation ( 2.70+/-0.35 versus 7.75+/-0.60-fold increase ; P=0.00007 ) , and a decrease in cytokine expression ( P < 0.05 ) . CONCLUSIONS : We conclude that inhibition of O00206 with eritoran in an in situ murine model significantly reduces MI/R injury and markers of an inflammatory response . Characterization of DB05005 -B , an orally bioavailable antagonist of the P51686 chemokine receptor , for treatment of inflammatory bowel disease . The chemokine system represents a diverse group of G protein-coupled receptors responsible for orchestrating cell recruitment under both homeostatic and inflammatory conditions . Chemokine receptor 9 ( P51686 ) is a chemokine receptor known to be central for migration of immune cells into the intestine . Its only ligand , O15444 , is expressed at the mucosal surface of the intestine and is known to be elevated in intestinal inflammation . To date , there are no reports of small-molecule antagonists targeting P51686 . We report , for the first time , the discovery of a small molecule , DB05005 -B , which is an orally bioavailable , selective , and potent antagonist of human P51686 . DB05005 -B inhibited P51686 -mediated Ca(2+) mobilization and chemotaxis on Molt-4 cells with IC(50) values of 5.4 and 3.4 nM , respectively . In the presence of 100 % human serum , DB05005 -B inhibited P51686 -mediated chemotaxis with an IC(50) of 33 nM , and the addition of α1-acid glycoprotein did not affect its potency . DB05005 -B inhibited chemotaxis of primary P51686 -expressing cells to O15444 with an IC(50) of 6.8 nM . DB05005 -B was an equipotent inhibitor of O15444 -directed chemotaxis of both splice forms of P51686 ( CCR9A and CCR9B ) with IC(50) values of 2.8 and 2.6 nM , respectively . DB05005 -B also inhibited mouse and rat P51686 -mediated chemotaxis . Inhibition of P51686 with DB05005 -B results in normalization of Crohn 's disease such as histopathology associated with the P01375 (ΔARE) mice . Analysis of the plasma level of drug associated with this improvement provides an understanding of the pharmacokinetic/pharmacodynamic relationship for P51686 antagonists in the treatment of intestinal inflammation . Q00613 inhibits expression of P05231 through activating transcription factor 3 . The febrile response is a complex physiological reaction to disease , including a cytokine-mediated increase in body temperature and the activation of inflammatory systems . Fever has beneficial roles in terms of disease prognosis , partly by suppressing the expression of inflammatory cytokines . However , the molecular mechanisms underlining the fever-mediated suppression of inflammatory gene expression have not been clarified . In this study , we showed that heat shock suppresses LPS-induced expression of P05231 , a major pyrogenic cytokine , in mouse embryonic fibroblasts and macrophages . Q00613 ( Q00613 ) activated by heat shock induced the expression of activating transcription factor ( P39905 ) 3 , a negative regulator of P05231 , and P18847 was necessary for heat-mediated suppression of P05231 , indicating a fever-mediated feedback loop consisting of Q00613 and P18847 . A comprehensive analysis of inflammatory gene expression revealed that heat pretreatment suppresses LPS-induced expression of most genes ( 86 % ) , in part ( 67 % ) via P18847 . When Q00613 -null and P18847 -null mice were injected with LPS , they expressed much higher levels of P05231 than wild-type mice , resulting in an exaggerated febrile response . These results demonstrate a novel inhibitory pathway for inflammatory cytokines . Indirubin-3'- ( 2,3 dihydroxypropyl ) -oximether ( E804 ) is a potent modulator of LPS-stimulated macrophage functions . Indirubin is a deep-red bis-indole isomer of indigo blue , both of which are biologically active ingredients in Danggui Longhui Wan , an ancient Chinese herbal tea mixture used to treat neoplasia and chronic inflammation and to enhance detoxification of xenobiotics . Multiple indirubin derivatives have been synthesized and shown to inhibit cyclin-dependent kinases ( CDKs ) and glycogen-synthase kinase ( GSK-3β ) with varying degrees of potency . Several indirubins are also aryl hydrocarbon receptor ( P35869 ) agonists , with P35869 -associated activities covering a wide range of potencies , depending on molecular structure . This study examined the effects of indirubin-3'- ( 2,3 dihydroxypropyl ) -oximether ( E804 ) , a novel indirubin with potent P40763 inhibitory properties , on basal and LPS-inducible activities in murine RAW264.7 macrophages . Using a focused commercial qRT-PCR array platform ( SuperArray® ) , the effects of E804 on expression of a suite of genes associated with stress and toxicity were determined . Most genes up-regulated by LPS treatment were suppressed by E804 ; including LPS-induced expression of pro-inflammatory cytokines and receptors , apoptosis control genes , and oxidative stress response genes . Using qRT-PCR as a follow up to the commercial arrays , E804 treatment suppressed LPS-induced P35354 , P35228 , P05231 and P22301 gene expression , though the effects on P35228 and P35354 protein expression were less dramatic . E804 also inhibited LPS-induced secretion of P05231 and P22301 . Functional endpoints , including P35228 and lysozyme enzymatic activity , phagocytosis of fluorescent latex beads , and intracellular killing of bacteria , were also examined , and in each experimental condition E804 suppressed activities . Collectively , these results indicate that E804 is a potent modulator of pro-inflammatory profiles in LPS-treated macrophages . Positive correlation between galactose-1-phosphate uridyltransferase ( P07902 ) and DB03501 -4'-epimerase ( Q14376 ) activities . OBJECTIVES : The aim of our study was to determine whether the activities of galactose-1-phosphate uridyltransferase and DB03501 -4'-epimerase are correlated , and in what way they may influence one another . DESIGN AND METHODS : Enzyme activities were measured in red blood cells from 214 individuals . RESULTS : A statistically significant ( p < 0.001 ) positive correlation was observed between P07902 and Q14376 activities . CONCLUSIONS : Our results suggest that P07902 and Q14376 activities are correlated and that Q14376 activity has a greater impact on P07902 activity than vice versa . Inhibition of cyclin-dependent kinases , GSK-3beta and CK1 by hymenialdisine , a marine sponge constituent . BACKGROUND : Over 2000 protein kinases regulate cellular functions . Screening for inhibitors of some of these kinases has already yielded some potent and selective compounds with promising potential for the treatment of human diseases . RESULTS : The marine sponge constituent hymenialdisine is a potent inhibitor of cyclin-dependent kinases , glycogen synthase kinase-3beta and casein kinase 1 . DB02950 competes with DB00171 for binding to these kinases . A P24941 -hymenialdisine complex crystal structure shows that three hydrogen bonds link hymenialdisine to the Glu81 and Leu83 residues of P24941 , as observed with other inhibitors . DB02950 inhibits Q00535 /p35 in vivo as demonstrated by the lack of phosphorylation/down-regulation of Pak1 kinase in E18 rat cortical neurons , and also inhibits GSK-3 in vivo as shown by the inhibition of P46821 phosphorylation . DB02950 also blocks the in vivo phosphorylation of the microtubule-binding protein tau at sites that are hyperphosphorylated by GSK-3 and Q00535 /p35 in Alzheimer 's disease ( cross-reacting with Alzheimer's-specific AT100 antibodies ) . CONCLUSIONS : The natural product hymenialdisine is a new kinase inhibitor with promising potential applications for treating neurodegenerative disorders . P19957 kinase/AKT pathway as a therapeutic target in multiple myeloma . The development of novel therapies for multiple myeloma depends on a comprehensive understanding of the events leading to cellular proliferation and survival . Controlling pathways that regulate growth signals is an emerging and complementary approach to myeloma treatment . The PI3K/Akt pathway is a central gatekeeper for crucial cellular functions including adhesion , angiogenesis , migration and development of drug resistance . Established proteins and genes such as P42345 , p53 , NF-kappaB and Q92934 are all regulated through PI3K and Akt activation , making them attractive targets for broad downstream effects . Direct PI3K inhibition has demonstrated impressive tumor inhibition and regression in cell-line and animal models , and multiple agents including DB05210 are currently in clinical trials . Drugs such as perifosine that are specific for Akt are also in development . Combinations of these agents with existing therapies are rational approaches on the path to improving myeloma treatment . Genetics and irritable bowel syndrome : from genomics to intermediate phenotype and pharmacogenetics . PURPOSE : Familial aggregation and sibling pair studies suggest there is a genetic contribution to the development of irritable bowel syndrome ( IBS ) . The aim of this study was to review the evidence of genetics in IBS based on genetic epidemiology , studies of association with intermediate phenotypes and pharmacogenetics . RESULTS : Genetic association studies with IBS symptom phenotype have generally provided inconsistent results for many candidate genes investigated , such as P31645 , P16520 , and P22301 . There have been no genome-wide association studies in IBS to date . Studies of associations of candidate genes with intermediate phenotypes suggest associations with pathophysiological mechanisms of motor and sensory functions ; however , these results also require replication . Pharmacogenetics studies illustrate the potential of genetics to impact on response to therapy , as observed with P31645 and responses to the 5- Q9H205 antagonist alosetron and the Q13639 agonist , tegaserod . CONCLUSIONS : While the heritable component and genetics in the complex disorder of IBS are still poorly understood , studies of the associations of spontaneous genetic variations and altered functions may provide novel insights of the mechanisms contributing to the disease . Lack of effect of methimazole on dendritic cell ( DC ) function and DC-induced Graves ' hyperthyroidism in mice . In addition to the biochemical inhibition of thyroid hormone synthesis , antithyroid drugs including methimazole ( MMI ) may have immunosuppressive effect through inhibition of major histocompatibility complex ( MHC ) class I and II expressions on non-professional ( thyrocytes ) and professional ( macrophages and B cells ) antigen presenting cells ( APCs ) . Dendritic cells ( DCs ) are another professional APCs and very likely play the most important role in the primary immune response . Therefore , we focused in this study on evaluating the effect of MMI on DC function in mice . Bone marrow cells cultured with granulocyte macrophage colony stimulating factor and interleukin ( IL ) -4 expressed high levels of CD11c and moderate levels of MHC class II , both of which are widely used markers for DCs . In vitro incubation of this DC-containing cell population with 10 ( - 6 ) -10 ( - 4 ) M MMI for 2 days did not change basal- and maturation signal ( adenoviral infection and lipopolysaccharide ) -induced levels of the cell surface marker expressions such as MHC class I and II , P42081 , P25942 and DEC205 , and of proinflammatory cytokine P05231 release . Further we found that treatment of the DC-containing cell population with MMI did not influence the incidence of Graves ' hyperthyroidism and anti-thyrotropin receptor ( P16473 ) antibody titers in a mouse Graves ' model we have recently established with DCs infected with adenovirus expressing the P16473 A subunit . Although we can not completely exclude immunosuppressive effect of MMI on other immune cells , our data indicate that DCs do not appear to be the primary target for the immunosuppressive effect of MMI . Endogenous basic fibroblast growth factor is essential for cyclin E- P24941 activity in multiple external cytokine-induced proliferation of AIDS-associated Kaposi 's sarcoma cells : dual control of AIDS-associated Kaposi 's sarcoma cell growth and cyclin E- P24941 activity by endogenous and external signals . AIDS-associated Kaposi 's sarcoma ( KS ) cell , a key element for development of KS lesions , proliferates in response to external cytokines , such as oncostatin M , the soluble IL-6R- P05231 complex , P01375 , and IL-1beta . In addition , the KS cell-produced basic fibroblast growth factor ( P09038 ) was reported to function as an autocrine growth factor . However , little is known of the exact roles of these external growth factors and endogenous P09038 on proliferation of KS cells , and underlying intracellular events have remained to be defined . We obtained evidence that anti- P09038 Ab abolished growth of KS cells by preventing S phase entry of the cell cycle , even in the presence of the external growth factors . Blockade of the FGF action profoundly inhibited cyclin E expression and cyclin-dependent kinase-2 ( P24941 ) activity , but not D-type cyclin expression and P11802 activity . Exogenously added acidic FGF ( P05230 ) , which generated a rapid tyrosine phosphorylation of P11362 and P21802 on KS cells , reversed the inhibitory effects of anti- P09038 Ab . Thus , FGF actions are essential for cyclin E- P24941 activity and S phase entry . We also observed that the presence of external growth factors markedly induced cyclin E- P24941 activity and S phase entrance , while the addition of P05230 or P09038 alone was insufficient to induce these responses . All this evidence shows that integration of the activities of external growth factors and endogenous P09038 is required for full activation of cyclin E- P24941 activity and KS cell proliferation . 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 . Contrasting activities of thyrotropin receptor antibodies in experimental models of Graves ' disease induced by injection of transfected fibroblasts or deoxyribonucleic acid vaccination . The development of experimental models of autoimmune hyperthyroid Graves ' disease has proved a difficult challenge , but recently two novel methods have led to their successful development in mice . We describe our studies on replicating the adjuvant modified , human DB00024 receptor ( P16473 ) and major histocompatibility complex class II transfected fibroblast injection system , and the plasmid DNA vaccination method as models resembling the human disorder . The fibroblast injection model in female AKR/N ( H-2k ) mice led to 70 % of the animals developing thyroid-stimulating antibodies and their thyroid glands showed large goiters with histological features of thyroid cell activation characteristic of Graves ' glands . Consistent with the clinical homolog , there was no inflammatory cell infiltrate of the thyroid gland . Detailed studies on the anti- P16473 antibodies such as thyroid-stimulating blocking antibody , antibodies to the native P16473 by flow cytometry , and DB00024 -binding inhibiting Ig showed that they were heterogeneous and did not correlate with disease activity , thus resembling those present in patients with Graves ' disease . In contrast , the plasmid DNA vaccination model in female BALB/c ( H-2d ) mice led to the generation of low levels of anti- P16473 antibodies by flow cytometry , which were undetectable for thyroid-stimulating antibodies , DB00024 -stimulating blocking antibodies , and DB00024 -binding inhibiting Ig activity . Moreover , this model too was not accompanied by lymphocytic cell infiltration . The data demonstrate the high incidence of hyperthyroid disease induced in the adjuvant modified , transfected fibroblast model in AKR/N mice to allow pathological mechanisms of disease to be studied .

MH_train_1397

MH_train_1397

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interacts_with DB00762?

multiple_choice

Chewing rescues stress-suppressed hippocampal long-term potentiation via activation of histamine H1 receptor . We have previously found in rats that chewing , an active behavioral strategy to cope with a stressful situation , rescues long-term potentiation ( LTP ) in the hippocampus through activating stress-suppressed N-methyl-D-aspartate ( DB01221 ) receptor function . To further examine the mechanisms underlying this ameliorative effect of chewing , we studied the involvement of the histaminergic system , which has been shown to be activated by mastication , in the LTP of hippocampal slices of rats that were allowed to chew a wooden stick during exposure to immobilization stress . Chewing failed to rescue stress-suppressed LTP in the rats treated with histamine H1 receptor ( P35367 ) antagonist DB06691 ( 5 mg/kg , i.p. ) before exposure to stress , although administration of DB06691 did not affect LTP in naive rats and in stressed rats that did not chew . However , when DB06691 was administrated immediately after exposure to stress , chewing rescued LTP whose magnitude was statistically comparable to that in the rats that chewed without drug treatment . These results suggest that chewing-induced histamine release in the hippocampus and the subsequent H1 receptor activation may be essential to rescue stress-suppressed synaptic plasticity . p38 Q96HU1 kinase regulates stem cell apoptosis in human hematopoietic failure . Myelodysplastic syndromes ( P43034 ) are clonal stem cell disorders that lead to ineffective hematopoiesis and are common causes of low blood counts in the elderly . The exact molecular mechanisms regulating increased stem apoptosis in these disorders are not well defined . p38 MAPK activation is important in regulating the growth inhibitory signals of P01375 , TGF-beta and Interferons on human hematopoiesis . Our findings show that p38 MAPK is overactivated in myelodysplasia bone marrows and regulates hematopoietic stem cell apoptosis . Inhibition of p38 MAPK by genetic or pharmacologic means decreases apoptosis and stimulates in vitro hematopoiesis from primary P43034 hematopoietic progenitors . These studies point to the potential efficacy of selective p38alpha inhibitor , DB05412 , in human bone marrow failure . Endogenous expression of histamine H1 receptors functionally coupled to phosphoinositide hydrolysis in P13671 glioma cells : regulation by cyclic AMP . 1. The effects of histamine receptor agonists and antagonists on phospholipid hydrolysis in rat-derived P13671 glioma cells have been investigated . 2 . P35367 -stimulation caused a concentration-dependent increase in the accumulation of total [ 3H ] -inositol phosphates in cells prelabelled with [ 3H ] -myo-inositol . The rank order of agonist potencies was histamine ( EC50 = 24 microM ) > N alpha-methylhistamine ( EC50 = 31 microM ) > 2-thiazolylethylamine ( EC50 = 91 microM ) . 3 . The response to 0.1 mM histamine was antagonized in a concentration-dependent manner by the H1-antagonists , mepyramine ( apparent Kd = 1 nM ) and (+)-chlorpheniramine ( apparent Kd = 4 nM ) . In addition , (-)-chlorpheniramine was more than two orders of magnitude less potent than its (+)-stereoisomer . 4 . Elevation of intracellular cyclic AMP accumulation with forskolin ( 10 microM , EC50 = 0.3 microM ) , isoprenaline ( 1 microM , EC50 = 4 nM ) or rolipram ( 0.5 mM ) , significantly reduced the histamine-mediated ( 0.1 mM ) inositol phosphate response by 37 % , 43 % and 26 % respectively . In contrast , 1,9-dideoxyforskolin did not increase cyclic AMP accumulation and had no effect on the phosphoinositide response to histamine . 5 . These data indicate the presence of functionally coupled , endogenous histamine H1 receptors in P13671 glioma cells . Furthermore , the results also indicate that H1 receptor-mediated phospholipid hydrolysis is inhibited by the elevation of cyclic AMP levels in these cells . DB08915 , a balanced PPARα/γ agonist , has no clinically relevant pharmacokinetic interaction with high-dose atorvastatin or rosuvastatin . BACKGROUND : DB08915 , a dual Q07869 -α/γ agonist , combines the lipid benefits of fibrates and the insulin-sensitizing benefits of thiazolidinediones . OBJECTIVE : To investigate the pharmacokinetic effects of co-administration of atorvastatin or rosuvastatin with aleglitazar . RESEARCH DESIGN AND METHODS : In a two-cohort , open-label , randomised , three-period crossover study , 44 healthy subjects received once-daily oral doses of aleglitazar 300 μg , statin ( atorvastatin 80 mg or rosuvastatin 40 mg ) and aleglitazar co-administered with each statin for 7 days . Plasma concentrations of each drug were measured and pharmacokinetic parameters determined on day 7 in each period . MAIN OUTCOME MEASURES : Peak observed plasma concentration ( C(max) ) and total exposures ( AUC ( 0 - 24 ) ) of aleglitazar , atorvastatin and rosuvastatin . RESULTS : C(max) and AUC ( 0 - 24 ) to aleglitazar were similar , whether administered alone or in combination with a statin . Total exposure to either statin was unaffected by co-administration with aleglitazar . C(max) treatment ratios for both statins exceeded the conventional no-effect boundary ( 1.25 ) when administered with aleglitazar . CONCLUSIONS : Co-administration of aleglitazar with a statin does not alter the pharmacokinetic profile of either drug . Novel camptothecin analogues that circumvent Q9UNQ0 -associated drug resistance in human tumor cells . DB00762 ( 7-ethyl-10-[4-(1-piperidino)-1-piperidino]-carbonyloxycamptothecin ; CPT-11 ) is a widely used potent antitumor drug that inhibits mammalian P11387 ( Topo I ) ; however , overexpression of Q9UNQ0 ( Q9UNQ0 / Q9UNQ0 / Q9UNQ0 ) can confer cancer cell resistance to SN-38 , the active form of CPT-11 . We have recently demonstrated that plasma membrane vesicles prepared from Q9UNQ0 -overexpressing PC-6/ Q8WUX1 -5H cells transported SN-38 and its glucuronide conjugate in an DB00171 -dependent manner ( Nakatomi et al. , Biochem Biophys Res Commun 2001;288:827-32 ) . In the present study , we have characterized a total of 14 new camptothecin ( CPT ) analogues with respect to both the inhibition of Topo I and the substrate specificity of Q9UNQ0 . All of the tested CPT analogues , which have different substitutions at positions 10 and 11 , strongly inhibited the Topo I activity in a cell-free system , as did SN-38 . Their antitumor activities in the SN-38-resistant PC-6/ Q8WUX1 -5H2 cell line greatly varied , however , being correlated with intracellular accumulation levels . We have examined DB00171 -dependent transport of those CPT analogues by using plasma membrane vesicles prepared from both PC-6/ Q8WUX1 -5H2 cells and Q9UNQ0 -transfected P29320 -293 cells . Based on the substrate specificity of Q9UNQ0 thus evaluated , it is strongly suggested that CPT analogues with high polarity are good substrates for Q9UNQ0 and are therefore effectively extruded from cancer cells . In this context , to circumvent Q9UNQ0 -associated drug resistance , low-polarity CPT analogues are considered to be potent lead compounds . The present study provides a practical approach to discover new CPT-based drugs for the chemotherapy of drug-resistant human cancer . DB04873 ( SB 207266 ) , a selective Q13639 receptor antagonist , reduces serotonin potentiation of neurally-mediated contractile responses of human detrusor muscle . The aim of this study is to evaluate the potency of piboserod ( SB 207266 ) , a selective 5-HT(4) receptor antagonist , at inhibiting the 5-HT(4)-mediated potentiating effect of serotonin ( 5-HT ) on the neurally-mediated contractile responses of human detrusor strips to electrical field stimulations ( O43281 ) . Strips of human detrusor muscle were mounted in Krebs-HEPES buffer under a resting tension of 500 mg and O43281 ( 20 Hz , 1 ms duration at 300 mA for 5 s ) was applied continuously at 1 min intervals . After stabilization of the O43281 -induced contractions , concentration-response curves to 5-HT ( 0.1 nM-100 microM ) were constructed in the absence or presence of 1 or 100 nM of piboserod . The experiments were performed in the presence of methysergide ( 1 microM ) and ondansetron ( 3 microM ) to block 5HT(1)/5HT(2) and 5-HT(3) receptors , respectively . 5-HT potentiated the contractile responses to O43281 of human bladder strips in a concentration-dependent manner , with a maximum mean of 60.0+/-19.9 % of the basal O43281 -evoked contractions . DB04873 did not modify the basal contractions but concentration-dependently antagonized the ability of 5-HT to enhance bladder strip contractions to O43281 . In presence of 1 and 100 nM of piboserod , the maximal 5-HT-induced potentiations were reduced to 45.0+/-7.9 and 38.7+/-8.7 % , respectively . A mean apparent antagonist dissociation constant value ( K(B) ) of 0.56+/-0.09 nM was determined . These data show the ability of piboserod to antagonize with high potency the enhancing properties of 5-HT on neurally-mediated contractions of isolated human bladder strips . Therefore , the 5-HT(4) receptor might represent an attractive pharmacological target for the treatment of overactive bladder . [ Quantitative analysis of P11387 activity in human and rat glioma : characterization and mechanism of resistance to antitopoisomerase chemical , camptothecin-11 ] . DB00762 ( CPT-11 ) is a new derivation of camptothecin , a plant alkaloid antitumor agent . Previous studies indicated that antitumor activity of CPT-11 was mediated through interaction of the drugs with its target enzyme , P11387 ( topo I ) . In this study , we studied the relation between sensitivity to CPT-11 and topo I activity of glioma cells . Furthermore , we established CPT-11 resistant cell lines in order to elucidate potential mechanisms of drug resistance . A clear correlation between the sensitivities to CPT-11 and topo I activities in surgical glioma specimens was demonstrated . Activities of topo I in CPT-11 sensitive group ( IC50 values for CPT-11 ; < 50 micrograms/ml ) tended to be higher than those in CPT-11 resistant group ( IC50 values ; > or = 50 ) . Topo I activity may serve as a novel marker to predict the sensitivity of gliomas to topo inhibitors . CPT-11 resistance cell lines ( T98G/CPT-11 and P13671 ) respectively exhibit a 5.4- and 7.3-fold increase in resistance to CPT-11 . No differences in topo I activity and intracellular accumulation of CPT-11 were observed between parent and CPT-11 resistant lines . On the other hand , topo I from T98G/CPT-11 and P13671 /CPT-11 cells were at least 4- and 2-fold resistant to the inhibitory effect of the CPT-11 on the relaxation activity of topo I in comparison with their parent lines . This enzymological difference may be responsible for the resistance to CPT-11 . Discovery and role of methylidene imidazolone , a highly electrophilic prosthetic group . The elimination of ammonia from alpha-amino acids is a chemically difficult process . While the non-acidic beta-proton has to be abstracted , the much more acidic ammonium protons must remain untouched to maintain the leaving group ability of this positively charged group . DB00117 and phenylalanine ammonia-lyases ( P42357 and Q9P2V4 ) possess a catalytically essential electrophilic group which has been believed to be dehydroalanine for 30 years . Recently , the X-ray structure of P42357 has been solved . The electron density was not consistent with dehydroalanine but showed the presence of methylidene imidazolone ( Q9NP71 ) instead . The high electrophilicity of this prosthetic group as well as the geometry at the active site support a previously proposed mechanism involving a Friedel-Crafts-type attack at the aromatic ring of the substrate . Further biochemical evidence for this unprecedented electrophile-assisted ammonia elimination is also presented . Although no X-ray structure of Q9P2V4 has been published as yet , spectrophotometrical evidence for the presence of Q9NP71 has been provided . Finally , a chemical model for the Q9P2V4 reaction is described . Mouse vascular endothelium activates CD8+ T lymphocytes in a P33681 -dependent fashion . Despite several studies examining the contribution of allorecognition pathways to acute and chronic rejection of vascularized murine allografts , little data describing activation of alloreactive T cells by mouse vascular endothelium exist . We have used primary cultures of resting or P01579 -activated C57BL/6 ( H-2(b) ) vascular endothelial cells as stimulators and CD8(+) T lymphocytes isolated from CBA/J ( H-2(k) ) mice as responders . Resting endothelium expressed low levels of MHC class I , which was markedly up-regulated after activation with P01579 . It also expressed moderate levels of P33681 at a resting state and after activation . Both resting and activated endothelium were able to induce proliferation of unprimed CD8(+) T lymphocytes , with proliferation noted at earlier time points after coculture with activated endothelium . Activated endothelium was also able to induce proliferation of P16070 (low) naive CD8(+) T lymphocytes . Activated CD8(+) T lymphocytes had the ability to produce P01579 and P60568 , acquired an effector phenotype , and showed up-regulation of the antiapoptotic protein Bcl-x(L) . Treatment with P16410 -Ig led to marked reduction of T cell proliferation and a decrease in expression of Bcl-x(L) . Moreover , we demonstrate that nonhemopoietic cells such as vascular endothelium induce proliferation of CD8(+) T lymphocytes in a P33681 -dependent fashion in vivo . These results suggest that vascular endothelium can act as an P25054 for CD8(+) direct allorecognition and may , therefore , play an important role in regulating immune processes of allograft rejection . DB01628 in the treatment of chronic pain . For the many patients who suffer chronic pain , we seek the most effective anti-inflammatory drug with the least side-effect profile and the greatest long-term safety . DB01628 , a selective P35354 inhibitor , has been shown to be as effective as non-selective non-steroidal anti-inflammatory drugs in the management of chronic pain in rheumatoid arthritis and osteoarthritis , for periods of up to one year . Data on etoricoxib efficacy in chronic low back pain is beginning to emerge . The side-effect profile of etoricoxib suggests it is well tolerated with similar adverse effects to non-selective NSAIDs . Larger studies are awaited , to see whether superior gastrointestinal tolerability can be proven . Further work will be required to show that etoricoxib is safe in patients with pre-existing cardiovascular or gastrointestinal comorbidity , and the potentially confounding role of aspirin still needs to be elucidated . However , etoricoxib shows promise as a new and effective P35354 inhibitor in clinical practice . Identification of multiple rare variants associated with a disease . Identifying rare variants that are responsible for complex disease has been promoted by advances in sequencing technologies . However , statistical methods that can handle the vast amount of data generated and that can interpret the complicated relationship between disease and these variants have lagged . We apply a zero-inflated Poisson regression model to take into account the excess of zeros caused by the extremely low frequency of the 24,487 exonic variants in the Genetic Analysis Workshop 17 data . We grouped the 697 subjects in the data set as Europeans , Asians , and Africans based on principal components analysis and found the total number of rare variants per gene for each individual . We then analyzed these collapsed variants based on the assumption that rare variants are enriched in a group of people affected by a disease compared to a group of unaffected people . We also tested the hypothesis with quantitative traits Q1 , Q2 , and Q4 . Analyses performed on the combined 697 individuals and on each ethnic group yielded different results . For the combined population analysis , we found that P22309 , which was not part of the simulation model , was associated with disease liability and that P17948 , which was a causal locus in the simulation model , was associated with Q1 . Of the causal loci in the simulation models , P17948 and P35968 were associated with Q1 and O95497 was correlated with Q2 . No significant genes were associated with Q4 . These results show the feasibility and capability of our new statistical model to detect multiple rare variants influencing disease risk . Novel agents that potentially inhibit irinotecan-induced diarrhea . DB00762 ( CPT-11 , 7-ethyl-10-[4-(1-piperidino)-1-piperidino] carbonyloxycamptothecin ) has exhibited clinical activities against a broad spectrum of carcinomas by inhibiting P11387 ( Topo I ) . However , severe and unpredictable dosing-limiting toxicities ( mainly myelosuppression and severe diarrhea ) hinder its clinical use . The latter consists of early and late-onset diarrhea , occurring within 24 hr or > or = 24 hr after CPT-11 administration , respectively . This review highlights novel agents potentially inhibiting CPT-11-induced diarrhea , which are designed and tested under guidance of disposition pathways and potential toxicity mechanisms . Early-onset diarrhea is observed immediately after CPT-11 infusion and probably due to the inhibition of acetylcholinesterase activity , which can be eliminated by administration of atropine . Late-onset diarrhea appears to be associated with intestinal exposure to SN-38 ( 7-ethyl-10-hydroxycamptothecin ) , the major active metabolite of CPT-11 , which may bind to Topo I and induce apoptosis of intestinal epithelia , leading to the disturbance in the absorptive and secretory functions of mucosa . CPT-11 and SN-38 may also stimulate the production of pro-inflammatory cytokines and prostaglandins ( PGs ) , thus inducing the secretion of Na(+) and Cl(-) . Early treatment of severe late-onset diarrhea with oral high-dose loperamide has decreased patient morbidity . Extensive studies have been conducted to identify other potential agents to ameliorate diarrhea in preclinical and clinical models . These include intestinal alkalizing agents , oral antibiotics , enzyme inducers , P-glycoprotein ( PgP ) inhibitors , cyclooxygenase-2 ( P35354 ) inhibitors , tumor necrosis factor-alpha ( P01375 ) inhibitors , or blockers of biliary excretion of SN-38 . Further studies are needed to identify the molecular targets associated with CPT-11 toxicity and safe and effective agents for alleviating CPT-11-induced diarrhea . Synthesis and characterization of the first fluorescent antagonists for human Q13639 receptors . Fluorescent antagonists for human 5-HT(4) receptors were synthesized based on ML10302 1 , a potent 5-HT(4) receptor agonist and on piperazine analogue 2 . These molecules were derived with three fluorescent moieties , dansyl , naphthalimide , and NBD ( 7-nitrobenz-2-oxa-1,3-diazol-4-yl ) , through alkyl chains . The synthesized molecules were evaluated in binding assays on the recently cloned human 5-HT(4(e)) receptor isoform stably expressed in P13671 glial cells with [(3)H]GR113808 as the radioligand . The affinity values depended upon the basal structure together with the alkyl chain length . The derivatives based on ML10302 were more potent ligands than the derivatives based on piperazine analogue . For ML10302-based ligands , dansyl and NBD derivatives attached through a chain length of one carbon atom 17a and 32 , respectively , led to affinities close to the affinity of ML10302 . The most potent compounds 17a , 28 , and 32 produced an inhibition of the 5-HT stimulated cyclic AMP synthesis in the same cellular system with nanomolar K(b) values . Fluorescent properties of 17a , 28 , and 32 were more particularly studied . Interactions of the fluorescent ligand 28 with the h5-HT(4(e)) receptor were indicated using h5-HT(4(e)) receptor transfected P13671 glial cell membranes and entire cells . Ligand 28 was also used in fluorescence microscopy experiments in order to label h5-HT(4(e)) receptor transfected P13671 glial cells , and subcellular localization of these receptors was more precisely determined using confocal microscopy . Lessons learned from the irinotecan metabolic pathway . DB00762 , a camptothecin analogue , is a prodrug which requires bioactivation to form the active metabolite SN-38 . SN-38 acts as a P11387 poison . DB00762 has been widely used in the treatment of metastatic colorectal cancer , small cell lung cancer and several other solid tumors . However , large inter-patient variability in irinotecan and SN-38 disposition , as well as severe but unpredictable diarrhea limits the clinical potential of irinotecan . Intense clinical pharmacology studies have been conducted to elucidate its complicated metabolic pathways and to provide scientific rationale in defining strategies to optimize drug therapy . DB00762 is subjected to be shunted between P08684 mediated oxidative metabolism to form two inactive metabolites P25054 or NPC and tissue carboxylesterase mediated hydrolysis to form SN-38 which is eventually detoxified via glucuronidation by P22309 to form SN-38G . The pharmacology of this compound is further complicated by the existence of genetic inter-individual differences in activation and deactivation enzymes of irinotecan ( e.g. , P08684 , P20815 , P22309 ) and sharing competitive elimination pathways with many concomitant medications , such as anticonvulsants , St . John 's Wort , and ketoconazole . Efflux of the parent compound and metabolites out of cells by several drug transporters ( e.g. , Pgp , Q9UNQ0 , MRP1 , Q92887 ) also occurs . This review highlights the latest findings in drug activation , transport mechanisms , glucuronidation , and CYP3A-mediated drug-drug interactions of irinotecan in order to unlock some of its complicated pharmacology and to provide ideas for relevant future studies into optimization of this promising agent . Mechanisms of atrial-selective block of Na⁺ channels by ranolazine : I . Experimental analysis of the use-dependent block . Atrial-selective inhibition of cardiac Na(+) channel current ( I(Na) ) and I(Na)-dependent parameters has been shown to contribute to the safe and effective management of atrial fibrillation . The present study examined the basis for the atrial-selective actions of ranolazine . Whole cell I(Na) was recorded at 15°C in canine atrial and ventricular myocytes and in human embryonic kidney ( P29320 ) -293 cells expressing Q14524 . Tonic block was negligible at holding potentials from -140 to -100 mV , suggesting minimal drug interactions with the closed state . Trains of 40 pulses were elicited over a range of holding potentials to determine use-dependent block . Guarded receptor formalism was used to analyze the development of block during pulse trains . Use-dependent block by ranolazine increased at more depolarized holding potentials , consistent with an interaction of the drug with either preopen or inactivated states , but was unaffected by longer pulse durations between 5 and 200 ms , suggesting a weak interaction with the inactivated state . Block was significantly increased at shorter diastolic intervals between 20 and 200 ms . Responses in atrial and ventricular myocytes and in P29320 -293 cells displayed a similar pattern . DB00243 is an open state blocker that unbinds from closed Na(+) channels unusually fast but is trapped in the inactivated state . Kinetic rates of ranolazine interactions with different states of atrial and ventricular Na(+) channels were similar . Our data suggest that the atrial selectivity of ranolazine is due to a more negative steady-state inactivation curve , less negative resting membrane potential , and shorter diastolic intervals in atrial cells compared with ventricular cells at rapid rates . DB04901 signals B- Q9NZ71 cells and inhibits the activities of NF-κB-induced P25490 - and snail-resistant factors : mechanism of sensitization to apoptosis by chemoimmunotherapeutic drugs . DB04901 ( anti- P33681 monoclonal antibody ) is a primatized ( human IgG1 constant regions and cynomologus macaque variable regions ) monoclonal antibody that is currently in clinical trials . DB04901 inhibits tumor cell proliferation through possibly cell signaling-mediated effects . Thus , we hypothesized that galiximab may signal the tumor cells and modify intracellular survival/antiapoptotic pathways such as the NF-κB pathway . This hypothesis was tested using various P33681 (+) Burkitt B- Q9NZ71 ( non-Hodgkin lymphomas ) cell lines as models . Treatment of B- Q9NZ71 cells with galiximab ( 25-100 μg/mL ) resulted in significant inhibition of NF-κB activity and its target resistant factors such as P25490 , Snail , and Bcl-2/Bcl-XL . Treatment of B- Q9NZ71 cells with galiximab sensitized the tumor cells to both DB00515 ( DB00515 ) - and P50591 -induced apoptosis . The important roles of P25490 - and Snail-induced inhibition by galiximab in the sensitization to CCDP and P50591 were corroborated following transfection of Raji cells with P25490 or Snail short interfering RNA . The transfected cells were shown to become sensitive to both CCDP- and P50591 -induced apoptosis in the absence of galiximab . Furthermore , knockdown of P25490 or Snail inhibited Bcl-XL . The involvement of Bcl-XL inhibition in sensitization was corroborated by the use of the pan-Bcl-2 inhibitor 2MAM-3 whereby the treated cells were sensitive to both DB00515 - and P50591 -induced apoptosis . These findings show that galiximab inhibits the NF-κB/Snail/ P25490 /Bcl-XL circuit that regulates drug resistance in B- Q9NZ71 and in combination with cytotoxic drugs results in apoptosis . The findings also support the therapeutic application of the combination of galiximab and cytotoxic drugs in the treatment of drug-resistant P33681 -positive B-cell malignancies . Metronomic chemotherapy dosing-schedules with estramustine and temozolomide act synergistically with anti- P35968 antibody to cause inhibition of human umbilical venous endothelial cell growth . The effects of ' metronomic ' or extended chemotherapy dosing schedules ( ECS ) are mediated through poorly understood anti-angiogenic mechanisms . ECS combined with biological anti-angiogenic agents have produced promising pre-clinical results . MATERIALS AND METHODS : We have expanded the list of agents with an in vitro ECS profile to include the methylating agent temozolomide ( DB00853 ) and the anti-mitotic agent estramustine ( Estracyt ) . These agents were also combined with a specific anti-angiogenic inhibitor DB06101 and a non-specific agent with anti-angiogenic properties , Compound 5h . The in vitro HUVEC ECS model system was optimised and cell proliferation assays undertaken . RESULTS : As a single agent , estramustine inhibited endothelial cell proliferation with an IC50 of 4.5 microM and was active at 10-33 % of the maximum tolerated dose ( MTD ) from clinical schedules , whilst temozolomide had IC50 of 6.6 microM and was active at 1-6 % of MTD . In combination , significant synergy was seen with DB06101 in combination with either drug , whilst modest additive effects were observed with Compound 5h . None of the combinations resulted in significant cytotoxicity or apoptosis . DISCUSSION : The results show that ECS of temozolomide and estramustine can be significantly enhanced when combined with specific anti-angiogenic inhibitors in an in vitro HUVEC system . The O60656 enzyme is a peroxisome proliferator-activated receptor alpha and gamma target gene . Peroxisome proliferator-activated receptor ( Q07869 ) alpha and gamma are ligand-activated transcription factors belonging to the nuclear receptor family . Q07869 alpha mediates the hypolipidemic action of the fibrates , whereas Q07869 gamma is a receptor for the antidiabetic glitazones . In the present study , the UDP-glucuronosyltransferase ( P78381 ) 1A9 enzyme is identified as a Q07869 alpha and Q07869 gamma target gene . UGTs catalyze the glucuronidation reaction , which is a major pathway in the catabolism and elimination of numerous endo- and xenobiotics . Among the P22309 family enzymes , O60656 metabolizes endogenous compounds , including catecholestrogens , and xenobiotics , such as fibrates and to a lesser extent troglitazone . Treatment of human hepatocytes and macrophages and murine adipocytes with activators of Q07869 alpha or Q07869 gamma resulted in an enhanced O60656 expression and activity . In addition , disruption of the Q07869 alpha gene in mice completely abolished the Q07869 alpha agonist-induced O60656 mRNA and activity levels . A Q07869 response element was identified in the promoter of O60656 at positions -719 to -706 bp by transient transfection and electromobility shift assays . Considering the role of O60656 in catecholestrogen metabolism , Q07869 alpha and Q07869 gamma activation may contribute to the protection against genotoxic catecholestrogens by stimulating their inactivation in glucuronide derivatives . Furthermore , since O60656 is involved in the catabolism of fibrates , these results suggest that Q07869 alpha and Q07869 gamma may control the intracellular level of active fibrates . Glial cell factors and the outer blood retinal barrier . The retinal pigment epithelium is an important barrier to drug transport as well as contributing to the normal functioning of the photoreceptors . The contributions of glial cells in the retina to the maintenance and development of this barrier is important . There is evidence that retinal secreted factors play a role in the induction and maintenance of the outer blood retinal barrier . One possible source of such factors are the retinal glial cells , astrocytes and Müller cells , which may influence tight junction formation and maturation . The aim of this study was to examine the changes in the trans-epithelial resistance ( Q9NZ01 ) , as a measure of barrier integrity , on cell lines of epithelial origin ( ECV304 and ARPE-19 ) following co-culture with glial cell lines ( P13671 and Q9NP71 -M1 ) or with the addition of medium conditioned by these cells . One cell line , ECV304 , showed a significant increase in the Q9NZ01 in response to glial secreted factors whilst ARPE-19 did not . This finding suggests that ECV304 responds well to glial factors and may be useful for further studies of the factors that affect tight junction formation through glial cell induction in vitro .

MH_train_1398

MH_train_1398

MH_train_1398

interacts_with DB01050?

multiple_choice

Novel marine phenazines as potential cancer chemopreventive and anti-inflammatory agents . Two new ( 1 and 2 ) and one known phenazine derivative ( lavanducyanin , 3 ) were isolated and identified from the fermentation broth of a marine-derived Streptomyces sp . ( strain CNS284 ) . In mammalian cell culture studies , compounds 1 , 2 and 3 inhibited P01375 -α-induced NFκB activity ( IC₅₀ values of 4.1 , 24.2 , and 16.3 μM , respectively ) and LPS-induced nitric oxide production ( IC₅₀ values of > 48.6 , 15.1 , and 8.0 μM , respectively ) . PGE₂ production was blocked with greater efficacy ( IC₅₀ values of 7.5 , 0.89 , and 0.63 μM , respectively ) , possibly due to inhibition of cyclooxygenases in addition to the expression of P35354 . Treatment of cultured HL-60 cells led to dose-dependent accumulation in the subG1 compartment of the cell cycle , as a result of apoptosis . These data provide greater insight on the biological potential of phenazine derivatives , and some guidance on how various substituents may alter potential anti-inflammatory and anti-cancer effects . Compound FLZ inhibits lipopolysaccharide-induced inflammatory effects via down-regulation of the P50750 -IKK and P50750 -JNK/p38MAPK pathways in RAW264.7 macrophages . AIM : The aim of this study was to investigate the effect of the squamosamide derivative FLZ ( N-2-(4-hydroxy-phenyl)-ethyl-2-(2,5-dimethoxy-phenyl)-3-(3-methoxy-4-hydroxy-phenyl)-acrylamide ) on lipopolysaccharide ( LPS ) -induced inflammatory mediator production and the underlying mechanism in RAW264.7 macrophages . METHODS : RAW264.7 cells were preincubated with non-toxic concentrations of compound FLZ ( 1 , 5 , and 10 micromol/L ) for 30 min and then stimulated with 10 microg/L LPS . The production of nitric oxide ( NO ) , the expression of inducible nitric oxide synthase ( P35228 ) and cyclooxygenase 2 ( P35354 ) , and the activation of nuclear factor kappa-B ( NF-kappaB ) and mitogen-activated protein kinase ( MAPK ) pathways were examined . RESULTS : FLZ significantly inhibited the LPS-induced production of NO , as well as the expression of P35228 and P35354 at both the RNA and the protein levels in RAW264.7 cells . The LPS-induced increase in the DNA binding activity of NF-kappaB and activator protein 1 ( AP-1 ) , the nuclear translocation of NF-kappaB p65 , the degradation of the inhibitory kappaBalpha protein ( P25963 ) and the phosphorylation of P25963 , O15111 ( IKK ) alpha/beta , c-Jun NH(2)-terminal kinase ( JNK ) and p38 MAPKs were all suppressed by FLZ . However , the phosphorylation of extracellular signal-regulated kinase ( P29323 ) was not affected . Further study revealed that FLZ inhibited the phosphorylation of transforming growth factor-beta ( TGF-beta ) -activated kinase 1 ( TAK1 ) , which is an upstream signaling molecule required for IKKalpha/beta , JNK and p38 activation . CONCLUSION : FLZ inhibited the LPS-induced production of inflammatory mediators at least partly through the downregulation of the P50750 -IKK and P50750 -JNK/p38MAPK pathways . cDNA array analysis of cytokines , chemokines , and receptors involved in the development of TNBS-induced colitis : homeostatic role of P01282 . Crohn 's disease ( CD ) is a chronic inflammatory pathology of the intestine , characterized by diarrhea and weight loss . A healing effect of vasoactive intestinal peptide ( P01282 ) in the murine model of CD based on 2,4,6-trinitrobencene sulfonic acid ( TNBS ) administration has been previously shown . The aim of this work was to analyze the expression of several mediators related to the inflammatory cascade in colitic and P01282 -treated animals . With this aim , mice received either only TNBS or TNBS and P01282 treatment on alternate days . cDNA microarray analysis and real-time polymerase chain reaction were performed on total mRNA from colon to study the expression of a battery of proinflammatory molecules such as the enzyme P35354 , the chemokines P78423 , P48061 , O43927 , O95715 , P51681 , and P25025 , and the cytokines interleukin ( IL ) -1beta , IL-12 , Q14116 , P22301 , interferon-gamma , and P05112 . TNBS administration induced the expression of all the proinflammatory mediators studied , whereas P01282 treatment reduced their levels , increasing the anti-inflammatory P22301 and the TH2 cytokine P05112 , explaining its beneficial action through inhibition of the inflammatory/ Q8IXH7 response . These data describe not only the relation of several proinflammatory mediators to the development of TNBS colitis , reporting their time-course , but also show the beneficial action of P01282 in this model through complete blockage of the inflammatory cascade and recovery of the colon homeostasis , providing a potential new alternative for CD therapy . The effect of feeding system in the expression of genes related with fat metabolism in semitendinous muscle in sheep . The effect of feeding system on the expression of P06858 , Q13085 , P49327 , P15090 , O75907 , O00767 , Q92523 , P54646 , P41159 , P36956 , P37231 , Q07869 and P17676 genes in semitendinous muscle was studied . Forty-four single born male lambs of the Rasa Aragonesa breed , allocated to four different dietary treatments , were used : grazing alfalfa , grazing alfalfa with supplement for lambs , indoor lambs with grazing ewes and drylot . Significant differences were found in the expression of genes P06858 , Q13085 , P49327 , P15090 , Q92523 and O00767 . Genes related to adipogenesis ( P06858 , Q13085 , P49327 , P15090 , and O00767 ) are up-regulated in the intensive groups . In grazing groups Q92523 gene expression , related to β-oxidation process , is up-regulated . The relative expression of Q92523 was 1.54 fold higher in Q9UNN4 +S , and 0.43 and 0.37 fold lower in IND- GRE and IND , respectively . The results support the hypothesis that changes in fatty acid profile due to feeding system implicate changes in the mRNA expression level of genes related with fat metabolism . Feeding strategy is an important tool to manipulate intramuscular fatty acid profile in meat through altering gene expression of enzymes related with fat metabolism . The bovine 5' AMPK gene family : mapping and single nucleotide polymorphism detection . The DB00131 -activated protein kinase ( AMPK ) family is an ancient stress response system whose primary function is regulation of cellular DB00171 . Activation of AMPK , which is instigated by environmental and nutritional stresses , initiates energy-conserving measures that protect the cell by inhibition and phosphorylation of key enzymes in energy-consuming biochemical pathways . The seven genes that comprise the bovine AMPK family were mapped in cattle by using a radiation hybrid panel . The seven genes mapped to six different cattle chromosomes , each with a LOD score greater than 10.0 . Q13131 mapped to BTA 20 , P54646 and O43741 to BTA 3 , Q9Y478 to BTA 17 , P54619 to BTA 5 , Q9UGJ0 to BTA 4 , and Q9UGI9 to BTA 2 . Five of the seven genes mapped to regions expected from human/cattle comparative maps . O43741 and Q9UGI9 , however , have not been mapped in humans . We predict these genes to be located on HSA 1 and 2 , respectively . Additionally , one synonymous and one non-synonymous single nucleotide polymorphism ( SNP ) were detected in Q9UGI9 in Bos taurus cattle . In an effort to determine ancestral origins , various herds of mixed breed cattle as well as other ruminant species were characterized for sequence variation in this region of Q9UGI9 . Owing to the physiological importance of this gene family , we believe that its individual genes are candidate genes for conferring resistance to diseases in cattle . The concerted regulation of P47712 , P35354 , and lipocortin 1 expression by IL-1beta in A549 cells . The pro-inflammatory effects of IL-1beta have been linked to the induction of the enzyme P35354 . We now show that in addition to increasing the expression of P35354 , IL-1beta concomittantly decreased the expression of lipocortin 1 on the surface of A549 cells . Furthermore , cytosolic P04054 is concomittantly activated by phosphorylation-resulting in a stimulation of arachidonic acid and DB00917 release . All of these effects appear to be mediated via a common pathway of P98160 and PKC activation . Activation of P47712 is inhibited by dexamethasone in a lipocortin 1-dependent mechanism . We present a novel hypothesis whereby the effects of IL-1beta are not only due to activation of enzymes necessary for generation of eicosanoids but also to an inhibition of mechanisms that regulate the supply of arachidonic acid . Whole blood lead concentration and erythrocyte delta-aminolevulinic acid dehydratase ( P13716 ) activity in selected canine populations in Greece . In a total number of 275 dogs of various ages , sex and breed , blood lead concentrations ( O43927 ) and erythrocyte P13716 activity were measured . Sixty-six of the dogs were living in lead mining areas ( Group A ) , 157 in urban areas ( Group B ) and 52 in rural areas ( Group C ) of Greece . Mean O43927 differed significantly ( P < 0.05 ) between locations and were 326,97 and 68 micrograms/L , respectively . Mean P13716 activity was significantly different ( P < 0.05 ) only between Groups A and B as between groups A and C . A significant ( P < 0.05 ) negative correlation existed between O43927 and P13716 activity . A normal range of erythrocyte P13716 activity of 807-992 mumol/ DB02272 /LRBC/h was established for dogs . None of the 33 Group A dogs and 2 of the Group B dogs that had a O43927 of 350 micrograms/L presented clinical signs indicating acute or chronic lead intoxication . No erythrocyte basophilic stippling or large number of nucleated red blood cells were seen in the 30 dogs of Group A with O43927 > 350 micrograms/L . Activation of intestinal peroxisome proliferator-activated receptor-α increases high-density lipoprotein production . AIMS : Peroxisome proliferator-activated receptor ( Q07869 ) -α is a transcription factor controlling lipid metabolism in liver , heart , muscle , and macrophages . Peroxisome proliferator-activated receptor-α activation increases plasma HDL cholesterol and exerts hypotriglyceridaemic actions via the liver . However , the intestine expresses Q07869 -α , produces HDL and chylomicrons , and is exposed to diet-derived Q07869 -α ligands . Therefore , we examined the effects of Q07869 -α activation on intestinal lipid and lipoprotein metabolism . METHODS AND RESULTS : The impact of Q07869 -α activation was evaluated in term of HDL-related gene expression in mice , ex vivo in human jejunal biopsies and in Caco-2/TC7 cells . Apolipoprotein-AI/HDL secretion , cholesterol esterification , and trafficking were also studied in vitro . In parallel to improving plasma lipid profiles and increasing liver and intestinal expression of fatty acid oxidation genes , treatment with the dual Q07869 -α/δ ligand DB05187 resulted in a more pronounced increase in plasma HDL compared with fenofibrate in mice . DB05187 , but not fenofibrate , increased the expression of HDL production genes such as apolipoprotein-AI and DB00171 -binding cassette A1 transporter in murine intestines . A similar increase was observed upon Q07869 -α activation of human biopsies and Caco-2/TC7 cells . Additionally , HDL secretion by Caco-2/TC7 cells increased . Moreover , Q07869 -α activation decreased the cholesterol esterification capacity of Caco-2/TC7 cells , modified cholesterol trafficking , and reduced apolipoprotein-B secretion . CONCLUSION : Peroxisome proliferator-activated receptor-α activation reduces cholesterol esterification , suppresses chylomicron , and increases HDL secretion by enterocytes . These results identify the intestine as a target organ of Q07869 -α ligands with entero-hepatic tropism to reduce atherogenic dyslipidaemia . Potentiation of P01138 -induced neurite outgrowth in PC12 cells by papaverine : role played by P98160 -γ , IP3 receptors . DB01113 , an inhibitor of phosphodiesterase ( PDE ) 10A , is gaining attention for its potential in the treatment of neuropsychiatric diseases such as schizophrenia . However , the precise mechanisms underlying the putative neuroprotective/neurotrophic actions of papaverine remain unclear . Thus , we investigated the effects of papaverine on nerve growth factor ( P01138 ) -induced neurite outgrowth in PC12 cells . DB01113 potentiated P01138 -induced neurite outgrowth in PC12 cells in a concentration-dependent manner . In contrast , the selective Q9Y233 inhibitor MP-10 had no effect on P01138 -induced neurite outgrowth . The potentiation of P01138 -induced neurite outgrowth by papaverine was blocked by the P98160 -γ inhibitor U73122 . Furthermore , papaverine 's potentiation of P01138 -induced neurite outgrowth was also blocked by the co-administration of inositol 1,4,5-trisphosphate ( IP(3) ) receptor antagonists ( xestospongin C and 2-aminoethoxydiphenyl borate ( 2- Q9H4A4 ) ) and by reduced expression of IP(3) receptor gene ( i.e. , itpr1 and itpr3 ) by siRNA . Our findings suggest that papaverine could potentiate P01138 -induced neurite outgrowth , and that activation of P98160 -γ and IP(3) receptors might be involved in the mechanism underlying papaverine 's potentiation of neurite outgrowth in PC12 cells . Not all monoclonals are created equal - lessons from failed drug trials in Crohn 's disease . The recent success of the anti-integrin antibody DB09033 can barely conceal the fact that the biologics armamentarium in Crohn 's disease has barely evolved beyond P01375 blockers so far . This contrasts with other immune-related diseases considered mechanistically and genetically closely related , such as psoriasis and rheumatoid arthritis , where approved biologics target a variety of independent biological mechanisms . Several pharmacological assets that entered clinical development have proven ineffective , or less effective than originally anticipated . While blockade of Q16552 and its receptor via DB09029 and Brodalumab , respectively , worsened Crohn 's disease , the beneficial effect of IL-12/23 p40 blockade via Ustekinumab appeared confined to a subpopulation of Crohn 's disease patients who have previously failed on P01375 blockers . Clinical development of the IFNγ blocker DB05111 was stopped despite demonstrating some clinical benefit , while the T cell co-stimulation blocker DB01281 did not exhibit any hint towards efficacy in Crohn 's disease . Here I review results from these individual development programmes , and also reflect on the lack of efficacy of the P01375 blocker DB00005 . I will discuss aspects of individual trials that might have confounded their interpretation and highlight the evolution in primary and secondary endpoints that have contributed to increasing robustness of results obtained in recent years . Finally , I suggest that mechanistic studies in murine genetic models combined with exploratory immunological studies incorporated in early drug development may represent the key for identifying the next generation of successful pharmacological targets in Crohn 's disease . In vivo activation of N-methyl-D-aspartate receptors in the rat hippocampus increases prostaglandin E(2) extracellular levels and triggers lipid peroxidation through cyclooxygenase-mediated mechanisms . Cyclooxygenases ( P36551 ) are a family of enzymes involved in the biosynthesis of prostaglandin ( PG ) and thromboxanes . The inducible enzyme cyclooxygenase-2 ( P35354 ) is the major isoform found in normal brain , where it is constitutively expressed in neurons and is further up-regulated during several pathological events , including seizures and ischaemia . Emerging evidence suggests that P35354 is implicated in excitotoxic neurodegenerative phenomena . It remains unclear whether PGs or other products associated to P36551 activity take part in these processes . Indeed , it has been suggested that reactive oxygen species , produced by P36551 , could mediate neuronal damage . In order to obtain direct evidence of free radical production during P36551 activity , we undertook an in vivo microdialysis study to monitor the levels of PGE(2) and 8-epi- P49763 (2alpha) following infusion of N-methyl-D-aspartate ( DB01221 ) . A 20-min application of 1 mm DB01221 caused an immediate , MK-801-sensitive increase of both PGE(2) and 8-epi- P49763 (2alpha) basal levels . These effects were largely prevented by the specific cytosolic phospholipase A(2) ( cPLA(2) ) inhibitor arachidonyl trifluoromethyl ketone ( Q06187 ) , by non- selective P36551 inhibitors indomethacin and flurbiprofen or by the P35354 selective inhibitor NS-398 , suggesting that the DB01221 -evoked prostaglandin synthesis and free radical-mediated lipid peroxidation are largely dependent on P35354 activity . As several lines of evidence suggest that prostaglandins may be potentially neuroprotective , our findings support the hypothesis that free radicals , rather than prostaglandins , mediate the toxicity associated to P35354 activity . Effect of milk hydrolysates on inflammation markers and drug-induced transcriptional alterations in cell-based models . Nonsteroidal anti-inflammatory drugs ( NSAID ) are associated with gastrointestinal inflammation and subsequent damage to the intestinal tissue . Earlier studies in our laboratory have found that specific casein hydrolysates ( CH ) might be useful in the treatment of gastrointestinal wounds . The underlying mechanisms that support inflammation and wound healing are not completely understood , but transcriptional alterations may be used as markers for inflammation and wound healing . The bioactivity of 3 CH prepared by treatment of commercial casein with pepsin ( 60 min ) followed by corolase ( 0 , 10 , or 60 min ) were investigated in intestinal epithelial cells treated with the NSAID indomethacin . The bioactivity was evaluated as transcriptional alterations of transforming growth factor-β1 ( TGF-β1 ) , cyclooxygenase-2 ( P35354 ) , peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) and nuclear factor κB ( NFκB ) by real-time PCR . Furthermore , the effect of CH on lipopolysaccharide-induced inflammation was evaluated in macrophages by measuring PG E(2) levels . Casein hydrolysates treated with corolase for 10 or 60 min after pepsin treatment downregulated transcription of TGF-β1 and NFκB ( P < 0.05 ) compared with the hydrolysate treated with pepsin only . Hydrolysate prepared by corolase treatment for 60 min after pepsin hydrolysis downregulated transcription of P35354 ( P < 0.05 ) compared with hydrolysate treated with corolase for only 10 min whereas transcription of Q07869 -γ was not affected ( P > 0.05 ) . Additionally , the hydrolysate prepared by pepsin treatment only ( 0 min corolase ) had a pro-inflammatory effect on macrophages via PG E(2) stimulation ( P < 0.05 ) . In conclusion , CH produced by a combination of pepsin and corolase treatments downregulated the transcription levels of TGF-β1 , P35354 , and NFκB . Interrelationship between signal transduction pathways and 1,25(OH)2D3 in UMR106 osteoblastic cells . In this study , the interrelationship between signal transduction pathways and 1,25-dihydroxyvitamin D(3) [ 1,25(OH)2D3 ] action was examined in UMR106 osteoblastic cells . Treatment of these cells with 8-bromo- DB02527 ( 1 mM ) resulted in an upregulation of the vitamin D receptor ( P11473 ) and an augmentation in the induction by 1,25(OH)2D3 of DB00146 24-hydroxylase [ 24(OH)ase ] and osteopontin ( P10451 ) mRNAs as well as gene transcription . Transfection with constructs containing the vitamin D response element devoid of other promoter regulatory elements did not alter the DB02527 -mediated potentiation , suggesting that DB02527 -enhanced transcription is due , at least in part , to upregulation of P11473 . Treatment with phorbol ester [ 12-O-tetradecanoyl-phorbol-13-acetate ( TPA ) 100 nM ] , an activator of protein kinase C , significantly enhanced 1,25(OH)2D3-induced P10451 mRNA and transcription but had no effect on P11473 or on 24(OH)ase mRNA or transcription . Studies using P10451 promoter constructs indicate that TPA-enhanced P10451 transcription is mediated by an effect on the P10451 promoter separate from an effect on P11473 . Thus interactions with signal transduction pathways can enhance 1,25(OH)2D3 induction of 24(OH)ase and P10451 gene expression , and , through different mechanisms , changes in cellular phosphorylation may play a significant role in determining the effectiveness of 1,25(OH)2D3 on transcriptional control in cells expressing skeletal phenotypic properties . MEK inhibition in non-small cell lung cancer . P01116 mutations are the most common mutations in non-small cell lung cancer ( NSCLC ) with adenocarcinoma histology . P01116 mutations result in the activation of the RAF-MEK- P29323 pathway , and agents that target RAF-MEK- P29323 pathways have been investigated in P01116 mutant NSCLC . The two agents furthest in development are selumetinib and trametinib . DB08911 has greater binding for the Q02750 /2 allosteric site , and generally has superior pharmacokinetics . A randomized phase II trial of docetaxel with and without selumetinib revealed that the combination resulted numerically superior overall survival , and a statistically significant improvement in progression-free survival and objective response rate . However , a concerning rate of hospital admission , grade 3 or 4 neutropenia , and febrile neutropenia was observed with the combination . Trials have investigated MEK inhibitors as single agents and in combination with erlotinib , and the data do not support the further development . The activity of MEK inhibitors appears to be similar in patients with P01116 mutant and wild-type NSCLC suggesting P01116 mutation status is not a reliable biomarker for efficacy . It is possible that mutations of genes in addition to P01116 mutations impact the activity of MEK inhibitors , or specific subsets of P01116 mutations may be resistant or susceptible to MEK inhibition . Other potential explanations are gene amplifications , alternative RNA splicing of genes resulting in activation of their protein products , and deregulation of noncoding RNAs and consequent altered protein expression . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . EV71 induces P35354 expression via c-Src/ P09619 /PI3K/Akt/ Q8NFH3 / Q8TCB0 MAPK/AP-1 and NF-kappaB in rat brain astrocytes . Enterovirus 71 ( EV71 ) induces the expression of cyclooxgenase ( P36551 ) -2 served as a major neurotoxic factor in CNS injury . However , the mechanisms underlying EV71-initiated intracellular signaling pathways leading to P35354 expression remain unknown . Therefore , we investigated the mechanisms underlying EV71-induced P35354 expression and prostaglandin E(2) ( PGE(2) ) production in rat brain astrocytes ( RBA ) -1 , determined by Western blotting , RT-PCR , and promoter assay . Here , we reported that EV71-induced P35354 expression and PGE(2) production were attenuated by pretreatment with the inhibitors of c-Src ( P50391 ) , P09619 ( AG1296 ) , PI3K ( Wortmannin ) , Q02750 /2 ( PD98059 ) , NF-kappaB ( helenalin ) , and AP-1 ( Tanshinone ) and transfection with shRNA or siRNA of c-Src , P09619 , p85 , c-Jun , c-Fos , P27361 , or P28482 . We further observed that EV71-induced activation of Akt and Q8NFH3 / Q8TCB0 MAPK were mediated via c-Src and P09619 . Pretreatment with P50391 attenuated EV71-stimulated phosphorylation of Src , P09619 , Akt , and Q8NFH3 / Q8TCB0 MAPK . Inhibition of PI3K by Wortmannin attenuated EV71-induced Akt and Q8NFH3 / Q8TCB0 MAPK phosphorylation , but had no effect on P09619 phosphorylation , suggesting that P09619 is an upstream and Q8NFH3 / Q8TCB0 MAPK is a downstream component of PI3K/Akt in these responses . EV71-stimulated NF-kappaB translocation from the cytoplasm to the nucleus , P25963 degradation and NF-kappaB promoter activity were attenuated by pretreatment with helenalin , but not AG1296 , Wortmannin , and PD98059 . EV71-induced c-Jun mRNA expression was attenuated by pretreatment with PD98059 , AG1296 , or Wortmannin . These results demonstrate that in RBA-1 cells , EV71-induced P35354 expression associated with PGE(2) production is mediated through activation of c-Src/ P09619 /PI3K/Akt/ Q8NFH3 / Q8TCB0 MAPK to initiate the expression of AP-1 . Innate immune responses to Q9NP99 activation : overlap , divergence , and positive and negative cross-talk with bacterial lipopolysaccharide . Q9NP99 ( triggering receptor expressed on myeloid cells-1 ) is an orphan immunoreceptor expressed on monocytes , macrophages , and neutrophils . Q9NP99 associates with and signals via the adapter protein O43914 / O43914 , which contains an ITAM . Q9NP99 activation by receptor cross-linking has been shown to be proinflammatory and to amplify some cellular responses to TLR ligands such as bacterial LPS . To investigate the cellular consequences of Q9NP99 activation , we have characterized global gene expression changes in human monocytes in response to Q9NP99 cross-linking in comparison to and combined with LPS . Both Q9NP99 activation and LPS up-regulate chemokines , cytokines , matrix metalloproteases , and PTGS/ P35354 , consistent with a core inflammatory response . However , other immunomodulatory factors are selectively induced , including P10451 and P09603 ( i.e. , P09603 ) by Q9NP99 activation and IL-23 and P09919 ( i.e. , DB00099 ) by LPS . Additionally , cross-talk between Q9NP99 activation and LPS occurs on multiple levels . Although synergy in GM- P04141 protein production is reflected in commensurate mRNA abundance , comparable synergy in IL-1beta protein production is not . Q9NP99 activation also attenuates the induction of some LPS target genes , including those that encode IL-12 cytokine family subunits . Where tested , positive Q9NP99 outputs are greatly reduced by the PI3K inhibitor wortmannin , whereas this attenuation is largely PI3K independent . These experiments provide a detailed analysis of the cellular consequences of Q9NP99 activation and highlight the complexity in signal integration between ITAM- and TLR-mediated signaling . Synergistic roles of platelet-derived growth factor-BB and interleukin-1beta in phenotypic modulation of human aortic smooth muscle cells . The phenotype of smooth muscle cells ( SMCs ) plays an important role in vascular function in health and disease . We investigated the mechanism of modulation of SMC phenotype ( from contractile to synthetic ) induced by the synergistic action of a growth factor ( platelet-derived growth factor , DB00102 ) and a cytokine ( interleukin , IL-1beta ) . Human aortic SMCs grown on polymerized collagen showed high expression levels of contractile markers ( smooth muscle alpha-actin , myosin heavy chain , and calponin ) . These levels were not significantly affected by DB00102 and IL-1beta individually , but decreased markedly after the combined usage of DB00102 and IL-1beta . PDGF/IL-1beta costimulation also induced a sustained phosphorylation of Akt and P08133 ribosomal S6 kinase ( p70S6K ) . The effects of PDGF/IL-1beta costimulation on contractile marker expression and Akt and p70S6K phosphorylation were blocked by the phosphatidylinositol 3-kinase inhibitors wortmannin and LY294002 and by adenovirus expressing a dominant-negative Akt , and they were mimicked by constitutively active Akt . DB00102 /IL-1beta induced a sustained phosphorylation of PDGF receptor ( P09619 ) -beta and its association with IL-1 receptor ( IL-1R1 ) . Such activation and association of receptors were blocked by a P09619 neutralizing antibody ( AF385 ) , an IL-1R1 antagonist ( IL-1ra ) , as well as a specific inhibitor of P09619 phosphorylation ( AG1295 ) ; these agents also eliminated the DB00102 /IL-1beta-induced signaling and phenotypic modulation . DB00102 /IL-1beta inhibited the polymerized collagen-induced serum response factor DNA binding activity in the nucleus , and this effect was mediated by the P09619 /IL-1R1 association and phosphatidylinositol 3-kinase/Akt/p70S6K pathway . Our findings provide insights into the mechanism of SMC phenotypic modulation from contractile to synthetic , e.g. , in atherosclerosis . Three-year follow-up of treatment-naïve and previously treated patients with CLL and SLL receiving single-agent ibrutinib . DB09053 is an orally administered inhibitor of Q06187 that antagonizes B-cell receptor , chemokine , and integrin-mediated signaling . In early-phase studies , ibrutinib demonstrated high response rates and prolonged progression-free survival ( PFS ) in chronic lymphocytic leukemia ( CLL ) . The durable responses observed with ibrutinib relate in part to a modest toxicity profile that allows the majority of patients to receive continuous therapy for an extended period . We report on median 3-year follow-up of 132 patients with symptomatic treatment-naïve and relapsed/refractory CLL or small lymphocytic lymphoma . Longer treatment with ibrutinib was associated with improvement in response quality over time and durable remissions . Toxicity with longer follow-up diminished with respect to occurrence of grade 3 or greater cytopenias , fatigue , and infections . Progression remains uncommon , occurring primarily in some patients with relapsed del(17)(p13.1) and/or del(11)(q22.3) disease . Treatment-related lymphocytosis remains largely asymptomatic even when persisting > 1 year and does not appear to alter longer-term PFS and overall survival compared with patients with partial response or better . Collectively , these data provide evidence that ibrutinib controls CLL disease manifestations and is well tolerated for an extended period ; this information can help direct potential treatment options for different subgroups to diminish the long-term risk of relapse .

MH_train_1399

MH_train_1399

MH_train_1399

interacts_with DB01656?

multiple_choice

DB08895 . DB08895 ( CP-690,550 ; CP-690550 ; CP690550 ) , an orally active immunosuppressant , is being developed by Pfizer for the treatment of rheumatoid arthritis , inflammatory bowel disease , dry eyes , ankylosing spondylitis , psoriasis , psoriatic arthritis , and for the prevention of transplant rejection . DB08895 specifically inhibits Janus activated kinase 3 ( P52333 ) , which has a pivotal role in cytokine signal transduction that governs lymphocyte survival , proliferation , differentiation , and apoptosis . This review discusses the key development milestones and therapeutic trials of this drug . Administration of adenosine diphosphate-ribosyl transferase antagonist allows in vivo control of anti-dinitrophenyl response . DB03073 ( 3MB ) is one of a series of chemical inhibitors of the nuclear enzyme adenosine diphosphate ( ADP ) -ribosyl transferase ( P09874 ) , which has been shown to inhibit cell differentiation in vitro , but has no effect on differentiation independent proliferation . Treatment of mice with an optimal concentration of 3MB ( 20 mg/kg body weight ) at or 1 day after dinitrophenyl-keyhole limpet haemocyanin ( DNP-KLH ) immunisation reduced anti-DNP plaque-forming cell ( P27918 ) numbers to less than 10 % of those of control animals . The period for maximum P27918 suppression showed a narrow time window relative to immunisation , suggesting that in vivo , as in vitro , 3 MB was acting only on those lymphocytes differentiating in response to antigen . Experimental findings showed that it was possible to select for P27918 derived from different populations of DNP-responsive lymphocytes by adjusting the time of 3MB treatment relative to immunisation . When 3MB was used with antigen priming , the residual P27918 showed a lower average affinity than P27918 in mice treated with 3MB 3 days after priming , suggesting a differential selection of those lymphocytes responding either ' early ' or ' late ' in the primary immune response . A randomized , placebo-controlled study of the effects of the p38 MAPK inhibitor SB- DB05250 on blood biomarkers of inflammation in P48444 patients . The p38 mitogen-activated protein kinase ( MAPK ) signaling upregulates inflammation and is known to be increased in chronic obstructive pulmonary disease ( P48444 ) . The authors assessed the pharmacology of the novel p38 MAPK inhibitor SB- DB05250 using blood biomarkers in P48444 . Seventeen P48444 patients ( forced expiratory volume in 1 second 50 % -80 % predicted ) using short-acting bronchodilators participated in a double-blind , double-dummy , randomized , crossover study . Patients received single oral doses of SB- DB05250 7.5 mg and 25 mg , prednisolone 10 mg and 30 mg , and placebo . Blood was obtained predose and at 1 , 2 , 6 , and 24 hours postdose . Whole-blood sorbitol-induced phosphorylated ( p ) heat shock protein ( HSP ) 27 levels as a marker of p38 pathway activation and lipopolysaccharide-induced tumor necrosis factor ( P01375 ) -alpha production were assessed . Both doses of SB- DB05250 , but not prednisolone , significantly ( P < .0001 ) reduced weighted mean ( WM ) pHSP27 ( 0-6 hours ) by 58 % compared with placebo . WM P01375 production ( 0-24 hours ) was significantly reduced compared with placebo by SB- DB05250 25 mg ( 40 % , P = .005 ) and 7.5 mg ( 33.4 % , P = .02 ) , while prednisolone 30 mg and 10 mg caused 81.5 % and 58.2 % suppression , respectively ( both P < .0001 ) . SB- DB05250 inhibited the p38 MAPK pathway to a greater degree than prednisolone did . SB- DB05250 inhibited P01375 production . SB- DB05250 is a potent p38 MAPK inhibitor that potentially suppresses inflammation in P48444 . The unique alterations of hippocampus and cognitive impairment in chronic obstructive pulmonary disease . BACKGROUND : Cognitive impairment has been found in chronic obstructive pulmonary disease ( P48444 ) patients . However , the structural alteration of the brain and underlying mechanisms are poorly understood . METHODS : Thirty-seven mild-to-moderate P48444 patients , forty-eight severe P48444 patients , and thirty-one control subjects were recruited for cognitive test and neuroimaging studies . Serum levels of P04271 ,pulmonary function and arterial blood gas levels were also evaluated in each subject . RESULTS : The hippocampal volume was significantly smaller in P48444 patients compared to the control group . It is positively correlated with a mini mental state examination ( MMSE ) score , SaO2 in mild-to-moderate P48444 patients , the levels of PaO2 in both mild-to-moderate and severe P48444 patients . Higher P04271 concentrations were observed in mild-to-moderate P48444 patients , while the highest P04271 level was found in severe P48444 patients when compared to the control subjects . P04271 levels are negatively associated with MMSE in both mild-to-moderate and severe P48444 patients and also negatively associated with the hippocampal volume in the total P48444 patients . CONCLUSIONS : Hippocampal atrophy based on quantitative assessment by magnetic resonance imaging does occur in P48444 patients , which may be associated with cognitive dysfunction and the most prevalent mechanism accountable for hippocampal atrophy is chronic hypoxemia in P48444 . Higher serum P04271 levels may be peripheral biochemical marker for cognitive impairment in P48444 . Statistical epistasis and progressive brain change in schizophrenia : an approach for examining the relationships between multiple genes . Although schizophrenia is generally considered to occur as a consequence of multiple genes that interact with one another , very few methods have been developed to model epistasis . Phenotype definition has also been a major challenge for research on the genetics of schizophrenia . In this report , we use novel statistical techniques to address the high dimensionality of genomic data , and we apply a refinement in phenotype definition by basing it on the occurrence of brain changes during the early course of the illness , as measured by repeated magnetic resonance scans ( i.e. , an ' intermediate phenotype. ' ) The method combines a machine-learning algorithm , the ensemble method using stochastic gradient boosting , with traditional general linear model statistics . We began with 14 genes that are relevant to schizophrenia , based on association studies or their role in neurodevelopment , and then used statistical techniques to reduce them to five genes and 17 single nucleotide polymorphisms ( SNPs ) that had a significant statistical interaction : five for Q07343 , four for P78509 , four for Q15303 , three for Q9NRI5 and one for Q02297 . Five of the SNPs involved in these interactions replicate previous research in that , these five SNPs have previously been identified as schizophrenia vulnerability markers or implicate cognitive processes relevant to schizophrenia . This ability to replicate previous work suggests that our method has potential for detecting a meaningful epistatic relationship among the genes that influence brain abnormalities in schizophrenia . P52333 activity is necessary for phosphorylation of cytosolic phospholipase A2 and prostaglandin E2 synthesis by macrophages infected with Francisella tularensis live vaccine strain . Francisella tularensis , the causative agent of tularemia , modulates the host immune response to gain a survival advantage within the host . One mechanism of immune evasion is the ability of F. tularensis to induce the synthesis of the small lipid mediator prostaglandin E2 ( DB00917 ) , which alters the host T cell response making the host more susceptible to Francisella growth . DB00917 is synthesized by a tightly regulated biosynthetic pathway following stimulation . The synthesis of DB00917 begins with the liberation of arachidonic acid ( AA ) from membrane phospholipids by cytosolic phospholipase A2 ( P47712 ) . AA is subsequently converted to the unstable intermediate PGH2 by cyclooxygenase-2 ( P35354 ) , and PGH2 undergoes an isomerization reaction to generate DB00917 . Our objective was to identify F. tularensis-activated host signaling pathways that regulate the activity of the enzymes in the DB00917 -biosynthetic pathway . In this study , we show that P47712 , p38 mitogen-activated protein kinase ( MAPK ) , and P52333 ( P52333 ) signaling are necessary for F. tularensis-induced DB00917 production . Inhibition of P52333 activity reduced the phosphorylation of P47712 and P35354 protein levels . In addition , P52333 regulates P47712 phosphorylation independent of transcription . Moreover , p38 MAPK activity is required for F. tularensis-induced P35354 protein synthesis , but not for the phosphorylation of P47712 . This research highlights a unique signaling axis in which P52333 and p38 MAPK regulate the activity of multiple enzymes of the DB00917 -biosynthetic pathway in macrophages infected with F. tularensis . Immunohistochemical analysis of brain lesions using P04271 and glial fibrillary acidic protein antibodies in arundic acid- ( DB05343 ) treated stroke-prone spontaneously hypertensive rats . Stroke-prone spontaneously hypertensive rats ( SHRSP ) used as a model of essential hypertension cause a high incidence of brain stroke on the course of hypertension . Incidences and sizes of brain lesions are known to relate to the astrocyte activities . Therefore , relation between brain damage and the expression profile of the astrocytes was investigated with morphometric and immunohistochemical analyses using astrocyte marker antibodies of P04271 and glial fibrillary acidic protein ( P14136 ) with or without arundic acid administration , a suppressor on the activation of astrocytes . Arundic acid extended the average life span of SHRSP . An increase in brain tissue weight was inhibited concomitant with a lower rate of gliosis/hemosiderin deposit/scarring in brain lesions . P04271 - or P14136 -positive dot and filamentous structures were decreased in arundic acid-treated SHRSP , and this effect was most pronounced in the cerebral cortex , white matter , and pons , and less so in the hippocampus , diencephalon , midbrain , and cerebellum . Blood pressure decreased after administration of arundic acid in the high-dose group ( 100 mg/kg/day arundic acid ) , but not in the low-dose group ( 30 mg/kg/day ) . These data indicate that arundic acid can prevent hypertension-induced stroke , and may inhibit the enlargement of the stroke lesion by preventing the inflammatory changes caused by overproduction of the P04271 protein in the astrocytes . [ Current Topics on Vitamin D. Evolution of animals and vitamin D ] . DB00169 is already found in the early evolution of life , but essentially as inactive products of the photochemical reaction of 7-dehydrocholesterol . The full vitamin D endocrine system characterized by the specific vitamin D transport protein ( DBP ) , specific vitamin D-metabolizing CYP P450 enzymes , active vitamin D metabolites , 1α,25 ( OH ) 2D3 , specific vitamin D nuclear receptor ( P11473 ) , and fibroblast growth factor 23 ( Q9GZV9 ) became essential for maintaining calcium and bone homeostasis in terrestrial animals cope with the challenging of higher gravity and calcium-poor environment . The present review describes the story about the evolution of animals and vitamin D . Phosphodiesterase-4 influences the PKA phosphorylation status and membrane translocation of G-protein receptor kinase 2 ( P25098 ) in P29320 -293beta2 cells and cardiac myocytes . Membrane-recruitment of P25098 ( G-protein receptor kinase 2 ) provides a fundamental step in the desensitization process controlling GPCRs ( G-protein-coupled receptors ) , such as the beta2AR ( beta2-adrenergic receptor ) . In the present paper , we show that challenge of P29320 -293beta2 [ human embryonic kidney cells stably overexpressing the FLAG-tagged beta2AR-GFP ( green fluorescent protein ) ] cells with the beta-adrenoceptor agonist , isoprenaline , causes P25098 to become phosphorylated by PKA ( DB02527 -dependent protein kinase ) . This action is facilitated when DB02527 -specific DB05876 ( phosphodiesterase-4 ) activity is selectively inactivated , either chemically with rolipram or by siRNA ( small interfering RNA ) -mediated knockdown of Q07343 and Q08499 . DB05876 -selective inhibition by rolipram facilitates the isoprenaline-induced membrane translocation of P25098 , phosphorylation of the beta2AR by P25098 , membrane translocation of beta-arrestin and internalization of beta2ARs . DB05876 -selective inhibition also enhances the ability of isoprenaline to trigger the PKA phosphorylation of P25098 in cardiac myocytes . In the absence of isoprenaline , rolipram-induced inhibition of DB05876 activity in P29320 -293beta2 cells acts to stimulate PKA phosphorylation of P25098 , with consequential effects on P25098 membrane recruitment and P25098 -mediated phosphorylation of the beta2AR . We propose that a key role for DB05876 enzymes is : ( i ) to gate the action of PKA on P25098 , influencing the rate of P25098 phosphorylation of the beta2AR and consequential recruitment of beta-arrestin subsequent to beta-adrenoceptor agonist challenge , and ( ii ) to protect P25098 from inappropriate membrane recruitment in unstimulated cells through its phosphorylation by PKA in response to fluctuations in basal levels of DB02527 . Antiproliferative , Apoptotic , and Autophagic Activity of DB01270 , DB00112 , DB04895 , and DB08885 on Fibroblasts : Implication for Choroidal Neovascularization . Purpose. Choroidal neovascularization ( CNV ) is one of the most common complications of retinal diseases accompanied by elevated secretion of vascular endothelial growth factor ( P15692 ) . Intravitreal anti-VEGFs ( ranibizumab , bevacizumab , pegaptanib , and aflibercept ) can suppress neovascularization , decrease vascular permeability and CNV size , and , thereby , improve visual function . The antiproliferative , apoptotic , and autophagic effect of anti- P15692 drugs on fibroblasts found in CNVs has not been yet explored . Methods . Concentration-dependent cellular effects of the four anti-VEGFs were examined in L929 fibroblasts over a 5-day period . The cell survival , mitotic and polykaryocytic indices , the level of apoptosis and autophagy , and the cellular growth kinetics were all assessed . Results . The anti-VEGFs could inhibit the survival , mitotic activity , and proliferation as well as increase the cellular heterogeneity , apoptosis , and autophagy of the fibroblasts in a dose-dependent manner . Cellular growth kinetics showed ranibizumab to be less aggressive , but three other anti-VEGFs showed higher antiproliferative and apoptotic activity and expressed negative cellular growth kinetics . Conclusions . The antiproliferative , apoptotic , and autophagic activity of anti-VEGFs upon fibroblasts may explain the cellular response and the etiology of CNV involution in vivo and serve as a good study model for CNV in vitro . Bresol inhibits phosphodiesterase 4 gene expression and modulates the levels of select mediators of inflammation in human monocytic cells . Bresol-a poly-herbal formulation , has been reported to be effective against bronchial asthma and allergic rhinitis in children . In vivo studies have supported the anti-histaminic and anti-anaphylactic action of bresol . However , the mechanism of action of bresol in modulation of inflammation has not been studied at the cellular and molecular level . The present study was aimed to elucidate the mechanism(s) of action of bresol at the cellular and molecular levels , using human monocyte leukemia cells . The effects of bresol on phosphodiesterase 4B ( Q07343 ) gene expression were analyzed using human monocytic U937 leukemia cells . The ability of bresol to stimulate DB02527 formation in these cells , as well as its effects on mediators of inflammation like tumor necrosis factor-α ( TNFα ) , nitric oxide ( NO ) , and cycloxygenase-2 ( P35354 ) in lipopolysaccharide ( LPS ) -stimulated U937 cells , were also studied . The results here indicated that bresol exhibited potential anti-inflammatory properties by inhibiting LPS-induced Q07343 gene expression in the cells . Bresol also dose dependently activated DB02527 formation , and inhibited TNFα , NO , as well as P35354 formation in the LPS-stimulated cells . Based upon the results , we concluded that the reported anti-inflammatory activity of bresol might be attributed to its abilities to inhibit Q07343 and thus elevate DB02527 levels in human monocytes . The anti-inflammatory effects of bresol might also be a result of the capacity of bresol to modulate the formation of TNFα , NO , and P35354 in monocytes . The promotion of iron-induced generation of reactive oxygen species in nerve tissue by aluminum . DB01370 is suspected to play a role in several neurological disorders . Reactive oxygen species ( ROS ) lead to oxidative stress , which is thought to be a possible mechanism for neurological damage . Interactions between aluminum and iron , a known promoter of prooxidant events , were studied in cerebral tissues using a fluorescent probe to measure rates of generation of ROS . Al2(SO4)3 alone failed to stimulate ROS production over a wide range of concentrations ( 50-1000 microM ) . The aluminum-deferrioxamine chelate in the absence of iron could also not potentiate ROS formation . However , Al2(SO4)3 potentiated FeSO4-induced ROS , with a maximal effect at 10 microM Fe and 500 microM Al . DB01575 , a hydrated aluminum silicate , did not potentiate iron-induced ROS formation . Ferritin had a minor stimulatory effect on ROS generation , but this was not potentiated by the concurrent presence of Al2(SO4)3 . P02787 had no effect on basal rates of ROS generation , but when Al2(SO4)3 was also present , ROS production was enhanced . It is concluded that : 1 . There is a potentiation of iron-induced ROS by aluminum salts ; 2 . Free or complexed aluminum alone is not a key producer of ROS ; and 3 . High rates of ROS production are unlikely to be owing to the displacement by aluminum iron from its biologically sequestered locations . Regulation of Q9GZV9 expression in IDG-SW3 osteocytes and human bone by pro-inflammatory stimuli . Fibroblast growth factor-23 ( Q9GZV9 ) , produced by osteocytes , is the key physiological regulator of phosphate homeostasis . Sepsis patients often experience transient hypophosphataemia , suggesting the regulation of Q9GZV9 levels by pro-inflammatory factors . Here , we used the osteocyte-like cell line IDG-SW3 to investigate the effect of pro-inflammatory stimuli on Q9GZV9 production . In differentiated IDG-SW3 cultures , basal Fgf23 mRNA was dose-dependently up-regulated by pro-inflammatory cytokines P01375 , IL-1β and O43508 , and bacterial LPS . Similar effects were observed in human bone samples . P01375 - and IL-1β-induced Fgf23 expression was NF-κB-dependent . Conversely , mRNA encoding negative regulators of Q9GZV9 , Phex , Dmp1 and Enpp1 , were suppressed by P01375 , IL-1β , O43508 and LPS , independent of NF-κβ signalling . Galnt3 , the protein product of which protects intact Q9GZV9 protein from furin/furin-like proprotein convertase cleavage , increased in response to these treatments . C-terminal Q9GZV9 and intact Q9GZV9 protein levels also increased , the latter only in the presence of P09958 inhibitors , suggesting that enzymatic cleavage exerts critical control of active Q9GZV9 secretion by osteocytes . Our results demonstrate in principle that pro-inflammatory stimuli are capable of increasing osteocyte secretion of Q9GZV9 , which may contribute to hypophosphataemia during sepsis and possibly other inflammatory conditions . Constitutive activation of neuregulin/ P21860 signaling pathway in clear cell sarcoma of soft tissue . Clear cell sarcoma of soft tissue ( CCSST ) represents a highly malignant tumor of the musculoskeletal system that is characterized by the chromosomal translocation t(12;22)(q13;q12) of the Ewing sarcoma gene ( Q01844 ) and activating transcription factor 1 ( P18846 ) . In a former microarray expression study , we identified P21860 , a member of the epidermal growth factor receptor ( P00533 ) family , as a promising new diagnostic marker in the differential diagnosis of CCSST . Here we show that , besides ErbB3 , all CCSST cell lines ( n = 8 ) also express the ErbB2 receptor or the ErbB4 receptor , representing an adequate coreceptor of ErbB3 . The phosphorylation status of ErbB3 revealed these receptor pairs to be either constitutively activated in CCSST cells with high neuregulin-1 ( Q02297 ) expression ( n = 4 ) or activatable by exogenic Q02297 in cells showing low amounts of Q02297 mRNA ( n = 4 ) . Exogenous Q02297 stimulated the growth of a subset of CCSST cells but did not affect the kinetics of another subset . This difference was not strictly dependent on endogenous Q02297 expression ; however , the growth-inhibiting effect of the pan-ErbB tyrosine kinase inhibitor DB05424 or PD158780 clearly correlated with Q02297 expression indicating an autocrine growth stimulation loop which may constitute an interesting target of new therapeutic strategies in this tumor entity . Cross-talk between PKA-Cβ and p65 mediates synergistic induction of Q07343 by roflumilast and NTHi . Phosphodiesterase 4B ( Q07343 ) plays a key role in regulating inflammation . DB01656 , a phosphodiesterase (PDE)4-selective inhibitor , has recently been approved for treating severe chronic obstructive pulmonary disease ( P48444 ) patients with exacerbation . However , there is also clinical evidence suggesting the development of tachyphylaxis or tolerance on repeated dosing of roflumilast and the possible contribution of Q07343 up-regulation , which could be counterproductive for suppressing inflammation . Thus , understanding how Q07343 is up-regulated in the context of the complex pathogenesis and medications of P48444 may help improve the efficacy and possibly ameliorate the tolerance of roflumilast . Here we show that roflumilast synergizes with nontypeable Haemophilus influenzae ( NTHi ) , a major bacterial cause of P48444 exacerbation , to up-regulate PDE4B2 expression in human airway epithelial cells in vitro and in vivo . Up-regulated PDE4B2 contributes to the induction of certain important chemokines in both enzymatic activity-dependent and activity-independent manners . We also found that protein kinase A catalytic subunit β ( PKA-Cβ ) and nuclear factor-κB ( NF-κB ) p65 subunit were required for the synergistic induction of PDE4B2 . PKA-Cβ phosphorylates p65 in a DB02527 -dependent manner . Moreover , Ser276 of p65 is critical for mediating the PKA-Cβ-induced p65 phosphorylation and the synergistic induction of PDE4B2 . Collectively , our data unveil a previously unidentified mechanism underlying synergistic up-regulation of PDE4B2 via a cross-talk between PKA-Cβ and p65 and may help develop new therapeutic strategies to improve the efficacy of DB05876 inhibitor . Identification of the fused bicyclic 4-amino-2-phenylpyrimidine derivatives as novel and potent DB05876 inhibitors . 2-Phenyl-4-piperidinyl-6,7-dihydrothieno[3,4-d]pyrimidine derivative ( 2 ) was found to be a new DB05876 inhibitor with moderate Q07343 activity ( IC50=150 nM ) . A number of derivatives with a variety of 4-amino substituents and fused bicyclic pyrimidines were synthesized . Among these , 5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine derivative ( 18 ) showed potent Q07343 inhibitory activity ( IC50=25 nM ) . Finally , N-propylacetamide derivative ( 31b ) was determined as a potent inhibitor for both Q07343 ( IC50=7.5 nM ) and P01375 -α production in mouse splenocytes ( IC50=9.8 nM ) and showed good in vivo anti-inflammatory activity in the LPS-induced lung inflammation model in mice ( ID50=18 mg/kg ) . The binding mode of the new inhibitor ( 31e ) in the catalytic site of Q07343 is presented based on an X-ray crystal structure of the ligand-enzyme complex . Constellation of HCN channels and DB02527 regulating proteins in dendritic spines of the primate prefrontal cortex : potential substrate for working memory deficits in schizophrenia . Schizophrenia associates with impaired prefrontal cortical ( P27918 ) function and alterations in cyclic AMP ( DB02527 ) signaling pathways . These include genetic insults to disrupted-in-schizophrenia ( Q9NRI5 ) and phosphodiesterases ( PDE4s ) regulating DB02527 hydrolysis , and increased dopamine D1 receptor ( D1R ) expression that elevates DB02527 . We used immunoelectron microscopy to localize Q9NRI5 , P27815 , Q07343 , and D1R in monkey P27918 and to view spatial interactions with hyperpolarization-activated cyclic nucleotide-gated ( HCN ) channels that gate network inputs when opened by DB02527 . Physiological interactions between PDE4s and HCN channels were tested in recordings of P27918 neurons in monkeys performing a spatial working memory task . The study reveals a constellation of DB02527 -related proteins ( Q9NRI5 , P27815 , and D1R ) and HCN channels next to excitatory synapses and the spine neck in thin spines of superficial P27918 , where working memory microcircuits interconnect and spine loss is most evident in schizophrenia . In contrast , channels in dendrites were distant from synapses and DB02527 -related proteins , and were associated with endosomal trafficking . The data suggest that a DB02527 signalplex is selectively positioned in the spines to gate P27918 pyramidal cell microcircuits . Single-unit recordings confirmed physiological interactions between DB02527 and HCN channels , consistent with gating actions . These data may explain why P27918 networks are especially vulnerable to genetic insults that dysregulate DB02527 signaling . DB00644 antagonist in the management of prostate cancer . DB00044 -releasing hormone ( P01148 ) agonist therapy to induce medical castration has become the most common form of hormonal therapy for advanced and metastatic prostate cancer . When treatment is started , P01148 agonists initially stimulate the release of LH , causing a surge in serum testosterone that can precipitate a " flare " phenomenon or worsening of disease , particularly in patients with bone metastatic disease . DB00644 ( DB00644 ) receptor antagonism represents a newer approach to medical castration . DB00106 is a pure P30968 antagonist that is devoid of any P01148 agonist activity . Results from 1 phase II and 3 phase III clinical trials demonstrate that abarelix produces medical castration more quickly and without causing testosterone surge , as compared with P01148 agonists with or without a nonsteroidal antagonist . The safety profile in terms of adverse events is comparable between the 2 types of treatment , but the lack of testosterone surge with abarelix might confer a safety advantage by abolishing the risk of a disease flare . Levels of angiopoietins 1 and 2 in induced sputum supernatant in patients with P48444 . Pathological features of chronic obstructive pulmonary disease ( P48444 ) include lung vascular remodeling and angiogenesis . Q15389 ( Ang-1 ) , is an essential mediator of angiogenesis by establishing vascular integrity , whereas angiopoietin-2 ( Ang-2 ) acts as its natural inhibitor . We determined the levels of angiopoietins in sputum supernatants of patients with P48444 and investigated their possible association with mediators and cells involved in the inflammatory and remodeling process . Fifty-nine patients with P48444 , 25 healthy smokers and 20 healthy non-smokers were studied . All subjects underwent lung function tests , sputum induction for cell count identification and Ang-1 , Ang-2 , P15692 , TGF-β1 , P08253 , LTB4 , P10145 , albumin measurement in sputum supernatants . Airway vascular permeability ( AVP ) index was also assessed . Ang-2 levels were significantly higher in patients with P48444 compared to healthy smokers and healthy non-smokers [ median , interquartile ranges pg/ml , 267 ( 147-367 ) vs. 112 ( 67-171 ) and 98 ( 95-107 ) , respectively ; p < 0.001 ] . Regression analysis showed a significant association between Ang-2 levels and AVP index , P15692 , P10145 and P08253 levels in P48444 , the strongest being with P15692 . Our results indicate that induced sputum Ang-2 levels are higher in P48444 compared to healthy smokers and healthy non-smokers . Moreover , Ang-2 is associated with AVP , P10145 , P08253 , and P15692 , indicating a possible role for Ang-2 in the pathogenesis of the disease . Masked selection : a straightforward and flexible approach for the selection of binders against specific epitopes and differentially expressed proteins by phage display . Phage display is a well-established procedure to isolate binders against a wide variety of antigens that can be performed on purified antigens , but also on intact cells . As selection steps are performed in vitro , it is possible to focus the outcome of the selection on relevant epitopes by performing some additional steps , such as depletion or competitive elutions . However in practice , the efficiency of these steps is often limited and can lead to inconsistent results . We have designed a new selection method named masked selection , based on the blockade of unwanted epitopes to favor the targeting of relevant ones . We demonstrate the efficiency and flexibility of this method by selecting single-domain antibodies against a specific portion of a fusion protein , by selecting binders against several members of the seven transmembrane receptor family using transfected P29320 cells , or by selecting binders against unknown breast cancer markers not expressed on normal samples . The relevance of this approach for antibody-based therapies was further validated by the identification of four of these markers , P16422 , P02787 receptor 1 , Metastasis cell adhesion molecule , and Sushi containing domain 2 , using immunoprecipitation and mass spectrometry . This new phage display strategy can be applied to any type of antibody fragments or alternative scaffolds , and is especially suited for the rapid discovery and identification of cell surface markers . DB00133 residues 338 and 339 in the carboxyl-terminal tail of the type II gonadotropin-releasing hormone receptor are critical for beta-arrestin-independent internalization . Cloned mammalian type II DB00644 receptors have a carboxyl-terminal tail in contrast to the mammalian type I DB00644 receptors , which uniquely lack a carboxyl-terminal tail . Because this domain mediates internalization of many serpentine receptors , the internalization pathway of the marmoset monkey type II P30968 and the functional role of the carboxyl-terminal tail in internalization was studied . The internalization pathway of the type II P30968 was investigated in COS-1 cells by coexpressing G protein-coupled receptor kinases ( GRKs ) , dynamin-1 , and beta-arrestins . Internalization of the receptor requires GRKs and dynamin but does not require beta-arrestin . The type II P30968 can also internalize via beta-arrestin in the presence of exogenous beta-arrestins , suggesting that the receptor can use two distinct internalization pathways . Receptor internalization appears to occur via clathrin-coated pits and caveolae because disruption of either structure inhibits internalization . Progressive truncations of the carboxyl-terminal tail identified a region containing serine residues 338 and 339 as critical for receptor internalization . Substitution of these serine residues with alanine residues inhibited internalization , whereas substitutions with glutamic acid residues rescued internalization . Furthermore , a dominant-negative P25098 did not inhibit internalization of receptors having these serine substitutions , although it inhibited internalization of the wild-type receptor . These results together identify serine residues 338 and 339 in the carboxyl-terminal tail as critical for internalization of the type II P30968 and suggest that these residues undergo phosphorylation by GRKs . However , neither of these residues , nor the carboxyl-terminal tail , is required for beta-arrestin-dependent internalization .