MH_dev_341

Phosphodiesterase - 4 influences the PKA phosphorylation status and membrane translocation of G-protein receptor kinase 2 ( P25098 REA ) in P29320 REA - 293beta2 cells and cardiac myocytes . Membrane-recruitment of P25098 REA ( G-protein receptor kinase 2 ) provides a fundamental step in the desensitization process controlling GPCRs ( G-protein-coupled receptors ) , such as the beta 2AR ( beta 2 - adrenergic receptor ) . In the present paper , we show that challenge of P29320 REA - 293beta2 [ human embryonic kidney cells stably overexpressing the FLAG-tagged beta 2AR - GFP ( green fluorescent protein ) ] cells with the beta-adrenoceptor agonist , isoprenaline , causes P25098 REA 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 REA and Q08499 REA . DB05876 - selective inhibition by rolipram facilitates the isoprenaline-induced membrane translocation of P25098 REA , phosphorylation of the beta 2AR by P25098 REA , membrane translocation of beta-arrestin and internalization of beta 2ARs . DB05876 - selective inhibition also enhances the ability of isoprenaline to trigger the PKA phosphorylation of P25098 REA in cardiac myocytes . In the absence of isoprenaline , rolipram-induced inhibition of DB05876 activity in P29320 REA - 293beta2 cells acts to stimulate PKA phosphorylation of P25098 REA , with consequential effects on P25098 REA membrane recruitment and P25098 REA - mediated phosphorylation of the beta 2AR . We propose that a key role for DB05876 enzymes is : ( i ) to gate the action of PKA on P25098 REA , influencing the rate of P25098 REA phosphorylation of the beta 2AR and consequential recruitment of beta-arrestin subsequent to beta-adrenoceptor agonist challenge , and ( ii ) to protect P25098 REA from inappropriate membrane recruitment in unstimulated cells through its phosphorylation by PKA in response to fluctuations in basal levels of DB02527 .

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 REA 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 REA inhibitory activity ( IC50 = 25 nM ) . Finally , N-propylacetamide derivative ( 31b ) was determined as a potent inhibitor for both Q07343 REA ( IC50 = 7.5 nM ) and P01375 REA - 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 REA is presented based on an X-ray crystal structure of the ligand-enzyme complex .

Cross-talk between PKA-Cβ and p65 mediates synergistic induction of Q07343 REA by roflumilast and NTHi . Phosphodiesterase 4B ( Q07343 REA ) plays a key role in regulating inflammation . DB01656 SUB , a phosphodiesterase ( PDE ) 4 - selective inhibitor , has recently been approved for treating severe chronic obstructive pulmonary disease ( P48444 REA ) 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 REA up-regulation , which could be counterproductive for suppressing inflammation . Thus , understanding how Q07343 REA is up-regulated in the context of the complex pathogenesis and medications of P48444 REA 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 REA exacerbation , to up-regulate PDE 4B2 expression in human airway epithelial cells in vitro and in vivo . Up-regulated PDE 4B2 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 PDE 4B2 . PKA-Cβ phosphorylates p65 in a DB02527 - dependent manner . Moreover , Ser 276 of p65 is critical for mediating the PKA-Cβ-induced p65 phosphorylation and the synergistic induction of PDE 4B2 . Collectively , our data unveil a previously unidentified mechanism underlying synergistic up-regulation of PDE 4B2 via a cross-talk between PKA-Cβ and p65 and may help develop new therapeutic strategies to improve the efficacy of DB05876 inhibitor .

P00747 REA activator inhibitor - 1 promotes inflammatory process induced by cigarette smoke extraction or lipopolysaccharides in alveolar epithelial cells . P00747 REA activator inhibitor - 1 ( P05121 REA ) plays a role in regulating levels of some cytokines and cell migration in addition to its classic role in inhibiting fibrinolysis . P05121 REA levels in induced-sputum of chronic obstructive pulmonary disease ( P48444 REA ) patients were elevated significantly and correlated negatively with pulmonary function . To study the role of P05121 REA in inflammatory process in P48444 REA , the authors transfected alveolar epithelial cells ( AECs ) with siRNA-targeted P05121 REA and stimulated the cells by cigarette smoke extraction ( CSE ) or lipopolysaccharides ( LPS ) . The expression of inflammatory factors , interleukin - 8 ( P10145 REA ) and leukotriene B4 ( LTB 4 ) , and the monocyte migration were detected . The exposure to CSE or LPS induced the expression of P05121 REA , P10145 REA , and LTB 4 in AECs and monocyte migration to AECs . However , they were attenuated after transfection with siRNA-targeted P05121 REA . In conclusion , P05121 REA stimulates inflammation induced by CSE or LPS in AECs through up-regulation of inflammatory factors and promoting inflammatory cell migration . P05121 REA may play a proinflammatory role in the pathogenesis of P48444 REA .

DB06212 MEN , 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 REA 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 MEN 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 MEN is a promising pharmacological tool in the treatment of renal edema .

Reversal of dopamine D2 agonist-induced inhibition of ventral tegmental area neurons by Gq-linked neurotransmitters is dependent on protein kinase C , G protein-coupled receptor kinase , and dynamin . Dopaminergic neurons of the ventral tegmental area are important components of brain pathways related to addiction . Prolonged exposure of these neurons to moderate concentrations of dopamine ( DA ) decreases their sensitivity to inhibition by DA , a process called DA-inhibition reversal ( P30518 REA ) . P30518 REA is mediated by phospholipase C and conventional subtype of protein kinase C ( cPKC ) through concurrent stimulation of D2 and D1 - like DA receptors , or by D2 stimulation concurrent with activation of 5 - HT ( 2 ) or neurotensin receptors . In the present study , we further characterized this phenomenon by use of extracellular recordings in brain slices to examine whether P30518 REA is linked to G protein-coupled receptor kinase - 2 ( P25098 REA ) or dynamin by assessing P30518 REA in the presence of antagonists of these enzymes . P30518 REA was blocked by β - O14965 REA inhibitor , which inhibits P25098 REA , and by dynasore , which blocks dynamin . Reversal of inhibition by D2 agonist quinpirole was produced by serotonin ( 50 µM ) and by neurotensin ( 5-10 nM ) . Serotonin-induced or neurotensin-induced reversal was blocked by β - O14965 REA inhibitor , dynasore , or cPKC antagonist 5,6 , 7,13- tetrahydro - 13 - methyl - 5 - oxo - 12H - indolo [ 2,3- a ] pyrrolo [ 3,4 c ] carbazole - 12 - propanenitrile ( Gö6976 ) . This further characterization of P30518 REA indicates that cPKC , P25098 REA , and dynamin play important roles in the desensitization of D2 receptors . As drugs of abuse produce persistent increases in DA concentration in the ventral tegmental area , reduction of D2 receptor sensitivity as a result of drug abuse may be a critical factor in the processes of addiction .

Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) - only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 MEN users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) - exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12 - , 18 - , 24 - and 48 - h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 REA - positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 MEN who were treated with a single dose of mifepristone .

DB06643 MEN - - an emerging treatment for postmenopausal osteoporosis . IMPORTANCE OF THE FIELD : Osteoporosis is a common skeletal disease that is associated with an imbalance in bone remodeling . DB06643 MEN is an investigational fully human monoclonal antibody to receptor activator of NF-kappaB ligand ( O14788 REA ) , a cytokine member of the P01375 REA family that is the principal mediator of osteoclastic bone resorption . AREAS COVERED IN THIS REVIEW : The efficacy and safety of denosumab in the management of postmenopausal osteoporosis is evaluated by reviewing the published literature and presentations at scientific meetings through 2009 . WHAT THE READER WILL GAIN : This review focuses on the data on fracture risk reduction and safety endpoints of denosumab in the treatment of postmenopausal osteoporosis . TAKE HOME MESSAGE : In postmenopausal women with osteoporosis , denosumab ( 60 mg by subcutaneous injection every 6 months ) increased bone mineral density , reduced bone turnover markers , and reduced the risk of vertebral , hip and non-vertebral fractures . DB06643 MEN was well tolerated with a safety profile generally similar to placebo . It is a promising emerging drug for the prevention and treatment of postmenopausal osteoporosis .

Branched N-glycans and their implications for cell adhesion , signaling and clinical applications for cancer biomarkers and in therapeutics . Branched N-glycans are produced by a series of glycosyltransferases including N-acetylglucosaminyltransferases and fucosyltransferases and their corresponding genes . Glycans on specific glycoproteins , which are attached via the action of glycosyltransferases , play key roles in cell adhesion and signaling . Examples of this are adhesion molecules or signaling molecules such as integrin and P12830 REA , as well as membrane receptors such as the P01133 REA and TGFβ receptors . These molecules also play pivotal roles in the underlying mechanism of a variety of disease such as cancer metastasis , diabetes , and chronic obstructive pulmonary disease ( P48444 REA ) . Alterations in the structures of branched N-glycans are also hall marks and are useful for cancer biomarkers and therapeutics against cancer . This mini-review describes some of our recent studies on a functional glycomics approach to the study of branched N-glycans produced by N-acetylglucosaminyltransferases III , IV , V and IX ( Vb ) ( GnT-III , GnT-IV , V and IX ( Vb ) ) and fucosyltransferase 8 ( Fut 8 ) and their patho-physiological significance , with emphasis on the importance of a systems glycobiology approach as a future perspective for glycobiology .

Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 REA ( 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 MEN ( 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 REA ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 REA 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 .

Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 - length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E ( 2 ) ) binding was similar , E ( 2 ) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4 - hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E ( 2 ) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E ( 2 ) in lung adenocarcinoma cells from females , but not males . P06401 REA ( PR ) expression was increased by E ( 2 ) in two out of five adenocarcinoma cell lines from females , but none from males . E ( 2 ) decreased P12830 REA protein expression in some of the cell lines from females , as it did in MCF - 7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 REA expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 REA and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males .

Mucosal expression of basic fibroblastic growth factor , syndecan 1 and tumour necrosis factor-α in Crohn ' s disease in deep remission under treatment with anti-TNFα antibodies . BACKGROUND AND AIMS : Both inflammation and fibrosis may be detected in Crohn ' s disease ( CD ) . The molecular pattern of Basic Fibroblastic Growth Factor ( P09038 REA ) and P18827 REA ( SD1 ) expression is altered in stenosing CD , but we do not know what the behaviour of this teamwork factor is in CD in deep remission under treatment with anti-TNFα antibodies . Our aim was to compare the expression of P09038 REA , SD1 and P01375 REA - α in patients with CD in deep remission under treatment with DB00065 MENMAX DB00065 MEN ( IFX ) or DB00051 ( P00813 REA ) and a control group of patients with active CD . METHODS : We assessed the expression of P09038 REA , SD1 and P01375 REA - α in 10 patients with active CD and in 28 patients with CD in sustained deep remission for at least 6 months . All patients underwent surveillance colonoscopy with biopsies , while receiving maintenance therapy with IFX or P00813 REA . Analysis was conducted by real-time reverse transcriptase PCR ( RT-PCR ) in biopsy samples . RESULTS : We found that P09038 REA , SD1 and P01375 REA - α were significantly reduced under treatment with anti-TNFα versus controls ( p= 0.000 ) . P09038 REA and SD1 expression were similar between IFX and P00813 REA patients ( p= 0.335 and p= 0.289 , respectively ) , while P01375 REA - α was significantly under-expressed in P00813 REA patients ( p= 0.008 ) . CONCLUSIONS : P09038 REA , SD1 and P01375 REA - α are significantly reduced in CD patients in deep remission under treatment with anti-TNFα , likely as an expression of optimal control of inflammation . The significance of the P01375 REA - α under-expression in patients under treatment with P00813 REA with respect to those under treatment with IFX should be elucidated in further studies .