interacts_with DB09068?
Inhibition of HIV-associated reverse transcriptase by sugar-modified derivatives of thymidine 5'-triphosphate in comparison to cellular DNA polymerases alpha and beta . The sugar-modified dTTP analogues 2',3'-didehydro-2',3'-dideoxy-thymidine 5'-triphosphate ( ddeTTP ) , 2',3'-dideoxythymidine 5'-triphosphate ( ddTTP ) , 3'-fluorothymidine 5'-triphosphate ( FdTTP ) , and 3'-azidothymidine 5'-triphosphate ( N3dTTP ) are demonstrated to be very effective and selective inhibitors of the HIV-associated reverse transcriptase ( HIV-RT ) . This conclusion is based on a comparison of the ID50 values of the compounds for the HIV-RT ( ranging from 0.03 microM for ddeTTP to 0.1 microM for ddTTP ) and the cellular DNA polymerase alpha ( greater than 200 microM ) . P06746 is partially affected by N3dTTP ( ID50 = 31 microM ) and by the other analogues ( ID50 = 1-2.2 microM ) . FdTTP has proved as effective as N3dTTP ( ID50 = 0.05 microM ) in suppressing the HIV-RT activity . Kinetic analysis revealed for both dTTP analogues a competitive type of inhibition and the same P04264 values ( about 0.05 microM ) . Agonist-induced glycogenolysis in rabbit retinal slices and cultures . 1. The effects of different putative retinal transmitters and/or modulators on glycogenolysis in rabbit retinal slices and in retinal Müller cell cultures were examined . 2 . Incubation of rabbit retinal slices or primary retinal cultures ( either 3-5 day-old or 25-30 day-old ) in a buffer solution containing [ 3H ] -glucose resulted in the accumulation of newly synthesized [ 3H ] -glycogen . 3 . DB00368 ( NA ) , isoprenaline , vasoactive intestinal peptide ( P01282 ) , 5-hydroxytryptamine ( 5-HT ) and 8-hydroxy-dipropylaminetetralin ( 8-OH-DPAT ) stimulated the hydrolysis of this newly formed 3H-polymer . The potency order of maximal stimulations was : P01282 greater than NA greater than isoprenaline greater than 5-HT greater than 8-OH-DPAT . 4 . The putative retinal transmitters , dopamine , gamma-aminobutyric acid ( GABA ) , glycine and taurine and the muscarinic agonist carbachol ( CCh ) had no effect on [ 3H ] -glycogen content . 5 . The glycogenolytic effects of NA/isoprenaline and 5-HT/8-OH-DPAT appear to be mediated by beta-adrenoceptors and 5-HT1 receptors ( possibly P08908 ) , respectively while the P01282 -induced response involved another receptor subtype . 6 . Agonists which mediated [ 3H ] -glycogen hydrolysis also stimulated an increase in adenosine 3':5'-cyclic monophosphate ( cyclic AMP ) formation . Both responses are blocked to a similar extent by the same antagonists and so are probably mediated via the same receptor subtypes . Moreover , dibutyryl cyclic AMP ( db cyclic AMP ) promoted tritiated glycogen breakdown in the three retinal preparations . 7 . Not all receptors linked to cyclic AMP production however promote glycogenolysis . Dopamine and apomorphine stimulated cyclic AMP formation via D1-receptors without influencing glycogenolysis . These receptors are exclusively associated with neurones . DB09068 ( Lu AA21004 ) , a novel multimodal antidepressant , enhances memory in rats . The serotonergic system plays an important role in cognitive functions via various 5-HT receptors . DB09068 ( Lu AA21004 ) in development as a novel multimodal antidepressant is a 5- Q9H205 , P34969 and P28221 receptor antagonist , a P28222 receptor partial agonist , a P08908 receptor agonist and a 5-HT transporter ( 5-HTT ) inhibitor in vitro . Preclinical studies suggest that 5- Q9H205 and P34969 receptor antagonism as well as P08908 receptor agonism may have a positive impact on cognitive functions including memory . Thus vortioxetine may potentially enhance memory . We investigated preclinical effects of vortioxetine ( 1-10mg/kg administered subcutaneously [ s.c. ] ) on memory in behavioral tests , and on cortical neurotransmitter levels considered important in rat memory function . Contextual fear conditioning and novel object recognition tests were applied to assess memory in rats . Microdialysis studies were conducted to measure extracellular neurotransmitter levels in the rat medial prefrontal cortex . DB09068 administered 1h before or immediately after acquisition of contextual fear conditioning led to an increase in freezing time during the retention test . This mnemonic effect was not related to changes in pain sensitivity as measured in the hotplate test . Rats treated with vortioxetine 1h before training spent more time exploring the novel object in the novel object recognition test . In microdialysis studies of the rat medial prefrontal cortex , vortioxetine increased extracellular levels of acetylcholine and histamine . In conclusion , vortioxetine enhanced contextual and episodic memory in rat behavioral models . Further demonstration of its potential effect on memory functions in clinical settings is warranted . Pharmacologic suppression of hepatic O95477 activity in mice reduces high-density lipoprotein cholesterol levels but promotes reverse cholesterol transport . BACKGROUND : The role of hepatic O95477 ( O95477 ) in maintaining plasma high density lipoprotein cholesterol ( HDL-C ) levels is well established , but its role in reverse cholesterol transport ( RCT ) is unclear . DB01599 is a compound that reduces HDL-C levels but also reduces atherosclerosis in animal models and xanthomas in humans . The aim of the present study was to test the hypothesis that probucol inhibits hepatic O95477 activity , thereby reducing HDL-C levels but promoting RCT from macrophages . METHODS AND RESULTS : Wild-type ( WT ) C57BL/6 mice and scavenger receptor class B type I ( Q8WTV0 ) knockout mice were fed a chow diet containing 0.5 % probucol or normal chow for 2 weeks . In WT mice , probucol , despite decreasing HDL-C by > 80 % , effectively maintained macrophage RCT . In Q8WTV0 knockout mice , probucol also substantially reduced HDL-C but significantly increased macrophage RCT . Furthermore , probucol significantly enhanced the excretion of HDL-derived cholesterol into feces in both WT and Q8WTV0 knockout mice . DB01599 inhibited O95477 -dependent cholesterol efflux from mouse primary hepatocytes , and this effect was shown to be responsible for the effect of probucol on increasing the fecal excretion of HDL-derived cholesterol in vivo . CONCLUSIONS : We demonstrate that pharmacological inhibition of hepatic O95477 activity with probucol reduced HDL-C levels but promoted RCT through diversion of HDL-derived cholesterol from efflux back into plasma instead to excretion in the bile . These results explain the beneficial effects of probucol on atherosclerosis and xanthomas despite its HDL-lowering effects and suggest that inactivation of hepatic O95477 leads to increased RCT despite reducing plasma HDL-C levels . Serotonin : a local regulator in the mammary gland epithelium . Serotonin ( 5-hydroxytryptamine , 5-HT ) is a very simple molecule that plays key roles in complex communication mechanisms within the animal body . In the mammary glands , serotonin biosynthesis and secretion are induced in response to dilation of the alveolar spaces . Since its discovery several years ago , mammary 5-HT has been demonstrated to perform two homeostatic functions . First , serotonin regulates lactation and initiates the transition into the earliest phases of involution . Second , serotonin is a local signal that induces parathyroid hormone-related peptide ( P12272 ) , which allows the mammary gland to drive the mobilization of calcium from the skeleton . These processes use different receptor types , P34969 and 5-HT2 , respectively . In this review , we provide synthetic perspectives on the fundamental processes of lactation homeostasis and the adaptation of calcium homeostasis for lactation . We analyze the role of the intrinsic serotonin system in the physiological regulation of the mammary glands . We also consider the importance of the mammary serotonin system in pathologies and therapies associated with lactation and breast cancer . 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 . 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 . 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 . Identification and characterization of two parathyroid hormone-like molecules in zebrafish . Zebrafish ( Danio rerio ) have receptors homologous to the human PTH ( DB05829 ) / P12272 receptor ( Q03431 ) and PTH-2 receptor ( P49190 ) and an additional receptor ( PTH3R ) with high homology to the Q03431 . To find natural ligands for zPTH1R and zPTH3R , we searched the zebrafish genomic database and discovered two distinct regions that , when translated ( zPTH1 and zPTH2 ) , showed high homology to DB05829 . Isolation of cDNAs and determination of the intron/exon boundaries revealed genomic structures which were similar to known PTHs . Peptides consisting of the first 34 amino acids after the pre- and prosequences of the zebrafish PTHs ( zPTHs ) were synthesized and were shown to be fully active at the hPTH1R. zPTH2(1-34) was , however , approximately 30-fold less potent at the zPTH1R than DB05829 (1-34) , hPTHrP(1-36) , and zPTH1(1-34) . When tested with zPTH3R , zPTH1(1-34) and hPTHrP(1-36) showed similar potencies , whereas the potency of zPTH2(1-34) was moderately ( 3-fold ) reduced . To determine whether other fishes have multiple PTHs , we searched the genomic database of the Japanese pufferfish ( Takifugu rubripes ) and identified zPTH1 and zPTH2 homologs . Phylogenetic analysis showed that PTHs from zebrafish and pufferfish are more closely related to each other than to known mammalian PTH homologs or to P12272 and tuberoinfundibular peptide of 39 residues . This is consistent with evolution of two teleost PTH-like peptides occurring after the evolutionary divergence between fishes and mammals . Overall , the PTH system appears more complex in fishes than in mammals , providing evidence of continued evolution in nontetrapod species . The availability of multiple forms of fish PTH and their receptors provide additional tools for PTH ligand/receptor structure-function studies . DB00159 inhibits prostaglandin D2 generation by inhibiting cyclo-oxygenase-2 in cultured human mast cells . BACKGROUND : DB00159 ( EPA ) is catalysed by cyclo-oxygenase ( P36551 ) , as is arachidonic acid , and is a competitive inhibitor of arachidonate metabolism . OBJECTIVES : We examined the effect of EPA on prostaglandin ( PG ) D2 generation in the cultured human mast cells with IgE-anti-IgE challenge incubation . METHODS : Cultured human mast cells were incubated with EPA ( 1 micromol/L ) for 20 h , then challenged with anti-IgE incubation after treatment with IgE . At the same time , P36551 inhibitors were tested to identify P23219 and P35354 activity . PGD2 synthetic activity was also assayed in a cell-free homogenate of cultured mast cells with P36551 inhibitors and EPA . DB11320 in the culture medium and in cells was assayed with the HPLC-fluorescent method . PGD2 and PGD3 were assayed with gas chromatography-mass spectrometry and the stable isotope dilution method . RESULTS : Although EPA incubation did not affect histamine release by cultured human mast cells in response to IgE-anti-IgE challenge incubation , it did decrease PGD2 generation by inhibiting the P35354 pathway . In contrast , in the cell-free homogenate of cultured human mast cells , EPA inhibited both P23219 and P35354 activities . CONCLUSION : Pre-incubation with EPA primarily affects the P35354 pathway in cultured human mast cells and reduces PGD2 generation in response to IgE-anti-IgE challenge incubation . These findings suggest that P23219 and P35354 have different substrate flow systems in mast cells . They also suggest that endogenous EPA diet supplementation would reduce PGD2 production and could serve as an anti-inflammatory substrate in human mast cells . 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 . Sustained muscle expression of dystrophin from a high-capacity adenoviral vector with systemic gene transfer of T cell costimulatory blockade . Adenoviral vector ( Ad ) -mediated gene delivery of normal , full-length dystrophin to skeletal muscle provides a promising strategy for the treatment of Duchenne muscular dystrophy ( P11532 ) . However , cellular and humoral immune responses induced by vector gene transfer limit the application of this approach . Blockade of the costimulatory interaction between naïve T cells and antigen-presenting cells has proven to be a successful means to diminish immunity induced by gene transfer . In this study we explore the potential of supplementing dystrophin gene delivery to dystrophin-deficient Dmd mouse skeletal muscle with systemic gene delivery of DB01281 and CD40Ig molecules to effect costimulatory blockade . We found that systemic administration of a high-capacity Ad ( HC-Ad ) vector carrying murine DB01281 ( AdmCTLA4Ig ) either alone or codelivered with an HC-Ad vector carrying murine CD40Ig ( AdmCD40Ig ) provided sustained expression of recombinant full-length murine dystrophin from an HC-Ad vector carrying the dystrophin cDNA ( AdmDys ) . The level of AdmDys vector genomes remained stable in animals cotreated with systemic delivery of vectors carrying molecules to block costimulation . In addition , muscle P01730 (+) and CD8(+) T cell infiltrates and Th1 cytokine production by splenocytes were reduced . The production of neutralizing antibody against Ad vector was significantly inhibited in mice receiving systemic codelivery of both AdmCTLA4Ig and AdmCD40Ig , but not in the mice treated with AdmCTLA4Ig alone . The results suggested that coblockade of both P10747 / P33681 and P29965 / P25942 costimulatory pathways is required for effective inhibition of the Ad vector-induced humoral immune response in Dmd mice , whereas blockade of P10747 / P33681 alone by murine DB01281 would be sufficient for prolonged dystrophin expression in treated muscle . Effects of 1-beta-D-arabinofuranosylcytosine incorporation on elongation of specific DNA sequences by P06746 . DB00987 ( ara-C ) is an effective antileukemic agent which acts as an inhibitor of DNA synthesis . The precise mechanism responsible for this inhibitory effect , however , remains unclear . The present work has examined the effects of the triphosphate derivative , ara- P53007 , on purified P06746 . These studies were performed on M13 phage DNA templates of defined sequence . The results demonstrate that ara-C is incorporated into DNA by P06746 . The results also demonstrate that the incorporated ara-C residue acts as a relative chain terminator . Moreover , the relative chain terminating effects of ara-C are sequence specific . In this regard , DNA strand elongation was progressively slowed at sequences of two , three , and four contiguous sites for cytosine incorporation . We also demonstrate that the inhibitory effects of ara-C are reversed by competition with deoxycytidine-triphosphate for incorporation into the DNA strand . Taken together , these findings are consistent with structural differences of the incorporated arabinosyl moiety which alter reactivity of the 3'-terminus and thereby inhibit chain elongation . These findings also provide new insights regarding the inhibitory effects of ara-C on elongation of specific DNA sequences . DB00819 inhibits stimulated feline liver and gallbladder bicarbonate secretion . Bile acidification is a key factor in preventing calcium carbonate precipitation and gallstone formation . P00918 ( CA II ) , that is inhibited by acetazolamide , plays a role in regulation of the acid-base balance in many tissues . This study examines the effect of acetazolamide on secretin- and vasoactive intestinal peptide ( P01282 ) -stimulated gallbladder mucosal bicarbonate and acid secretion . Gallbladders in anaesthetized cats were perfused with a bicarbonate buffer bubbled with CO2 in air . In 20 experiments P01282 ( 10 microg kg(-1) h(-1) ) and in 10 experiments secretin ( 4 microg kg(-1) h(-1) ) were infused continuously intravenous ( i.v. ) . Hepatic bile and samples from the buffer before and after perfusion of the gallbladder were collected for calculation of ion and fluid transport . During basal conditions a continuous secretion of H+ by the gallbladder mucosa was seen . Intravenous infusion of vasoactive intestinal peptide ( P01282 ) and secretin caused a secretion of bicarbonate from the gallbladder mucosa ( P < 0.01 ) . This secretion was reduced by intraluminal ( i.l. ) acetazolamide ( P < 0.01 ) . Bile flow was enhanced by infusion of P01282 and secretin ( P < 0.01 ) but this stimulated outflow was not affected by i.v. acetazolamide . The presence of CA II in the gallbladder was demonstrated by immunoblotting . Biliary CA activity has an important function in the regulation of P01282 - and secretin-stimulated bicarbonate secretion across the gallbladder mucosa . Interactions between genetic variants of folate metabolism genes and lifestyle affect plasma homocysteine concentrations in the Boston Puerto Rican population . OBJECTIVE : To investigate genetic and lifestyle factors and their interactions on plasma homocysteine ( Hcy ) concentrations in the Boston Puerto Rican population . DESIGN : Cross-sectional study . Plasma concentrations of Hcy , folate , vitamin B12 and pyridoxal phosphate were measured , and genetic polymorphisms were determined . Data on lifestyle factors were collected in interviews . SETTING : A population survey of health and nutritional measures . SUBJECTS : A total of 994 Puerto Rican men and women residing in the Boston metropolitan area . RESULTS : Smoking status was positively associated with plasma Hcy . Genetic polymorphisms P42898 677C→T , Q04609 1561C→T , Q04609 rs647370 and Q96NT5 928A→G interacted significantly with smoking for Hcy . P42898 1298A→C ( P = 0·040 ) and Q96NT5 928A→G ( P = 0·002 ) displayed significant interactions with alcohol intake in determining plasma Hcy . Subjects with Q96NT5 928GG genotype had significantly higher plasma Hcy concentrations compared with carriers of the A allele ( AA+AG ; P = 0·030 ) among non-drinking subjects . When consuming alcohol , GG subjects had lower plasma Hcy levels compared with AA+AG subjects . Physical activity interacted significantly with Q99707 2756A→G in determining plasma Hcy ( P for interaction = 0·002 ) . Smoking interacted with physical activity for plasma Hcy ( P for interaction = 0·023 ) . CONCLUSIONS : Smoking and drinking were associated plasma Hcy concentrations . Genetic variants involved in folate metabolism further modify the effects of lifestyle on plasma Hcy . The genomic clone P08908 which resembles a beta-adrenergic receptor sequence encodes the P08908 receptor . The recent cloning of the complementary DNAs and/or genes for several receptors linked to guanine nucleotide regulatory proteins including the adrenergic receptors ( alpha 1 , alpha 2A , alpha 2B , beta 1 , beta 2 ) , several subtypes of the muscarinic cholinergic receptors , and the visual ' receptor ' rhodopsin has revealed considerable similarity in the primary structure of these proteins . In addition , all of these proteins contain seven putative transmembrane alpha-helices . We have previously described a genomic clone , P08908 , isolated by cross-hybridization at reduced stringency with a full length beta 2-adrenergic receptor probe . This clone contains an intronless gene which , because of its striking sequence resemblance to the adrenergic receptors , is presumed to encode a G-protein-coupled receptor . Previous attempts to identify this putative receptor by expression studies have failed . We now report that the protein product of the genomic clone , Q96NT5 , transiently expressed in monkey kidney cells has all the typical ligand-binding characteristics of the 5-hydroxytryptamine ( P08908 ) receptor . Interaction between endothelial differentiation-related factor-1 and calmodulin in vitro and in vivo . P62158 ( P62158 ) is the principal Ca(2+) receptor protein inside the cell . When activated by Ca(2+) , P62158 binds and activates target proteins , thus altering the metabolism and physiology of the cell . Under basal conditions , calcium-free P62158 binds to other proteins termed P62158 -binding proteins . Recently , we described endothelial differentiation-related factor ( P08476 ) -1 as a protein involved in the repression of endothelial cell differentiation ( Dragoni , I. , Mariotti , M. , Consalez , G. G. , Soria , M. , and Maier , J. A. M. ( 1998 ) J. Biol. Chem. 273 , 31119-31124 ) . Here we report that ( i ) O60869 binds P62158 in vitro and in vivo ; ( ii ) O60869 is phosphorylated in vitro and in vivo by protein kinase C ; and ( iii ) O60869 - P62158 interaction is modulated by the concentrations of Ca(2+) and by the phosphorylation of O60869 by protein kinase C both in vitro and in vivo . In addition , 12-O-tetradecanoylphorbol-13-acetate treatment of human umbilical vein endothelial cell stimulates the nuclear translocation of O60869 . On the basis of the high homology of O60869 with multiprotein bridging factor-1 , a transcriptional coactivator that binds TATA-binding protein ( P20226 ) , we also demonstrate that O60869 interacts with P20226 in vitro and in human endothelial cells . We hypothesize that O60869 serves two main functions in endothelial cells as follows : ( i ) to bind P62158 in the cytosol at physiologic concentrations of Ca(2+) and ( ii ) to act in the nucleus as a transcriptional coactivator through its binding to P20226 . 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 . P62158 interacts with DB00171 binding cassette transporter A1 to protect from calpain-mediated degradation and upregulates high-density lipoprotein generation . OBJECTIVE : To investigate the interaction of DB00171 -binding cassette transporter A1 ( O95477 ) with calmodulin in relation to its calpain-mediated degradation because many calpain substrates bind calmodulin to regulate cellular functions . METHODS AND RESULTS : The activity of O95477 is regulated through proteolysis by calpain . An immunoprecipitation and glutathione S-transferase pull-down assay revealed that O95477 directly binds calmodulin in a Ca(2+)-dependent manner . The cytoplasmic loop of O95477 contains a typical calmodulin binding sequence of 1-5-8-14 motifs ( 1245 to 1257 amino acids ) . The peptide of this region showed binding to calmodulin , and deletion of the 1-5-8-14 motif abolished this interaction . This motif is located near the O95477 Pro- DB00142 - DB00133 - DB00156 sequence , and the presence of calmodulin/Ca(2+) protected the peptides from proteolysis by calpain . The knockdown of calmodulin by a specific small and interfering RNA increased the degradation of O95477 and decreased O95477 protein and apolipoprotein A-I-mediated lipid release . Surprisingly , calmodulin inhibitor W7 increased calmodulin binding to O95477 and protected it from calpain-mediated degradation , consistent with our previous finding that this compound increased apolipoprotein A-I-mediated cell cholesterol release . CONCLUSIONS : P62158 directly binds and stabilizes O95477 in the presence of Ca(2+) and increases the generation of high-density lipoprotein . 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 . Early effects of topoisomerase I inhibition on RNA polymerase II along transcribed genes in human cells . We have determined the early effects of camptothecin and alpha-amanitin on genomic DNA-binding sites of RNA polymerase II ( RNAPII ) , TATA-binding protein ( P20226 ) , P11387 ( Top1 ) , and histone components in human transcribed loci by chromatin-immunoprecipitation ( ChIP ) . The two agents caused notably different alterations in active chromatin . DB04690 induced a specific reduction of RNAPII density at promoter pause sites and histone modifications suggesting an increased chromatin accessibility . alpha-Amanitin caused an accumulation of RNAPII at transcribed genes , a reduction of P20226 bound to chromatin and a less accessible chromatin structure . Interestingly , RNAPII reduction at promoter pause sites occurred within 5-10min of camptothecin treatment , and was not a response to replication-dependent DNA breaks . ChIP analyses of RNAPII along transcribed genes indicated that RNAPII levels were transiently increased at internal exons , and that camptothecin effects could be fully reversed by DRB , a cdk inhibitor . Top1 was found to be enriched in active chromatin , therefore suggesting that Top1 inhibition at the transcribed template and/or adjacent regulating regions immediately affects RNAPII at active genes . The findings are novel in vivo evidence of camptothecin effects on RNAPII bound to transcribing genomic regions , and are consistent with the hypothesis that Top1 activity can be involved in transcription regulation at the level of promoter clearance . 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 . P08908 receptor activation : short-term effects on the mRNA expression of the P08908 receptor and galanin in the raphe nuclei . Systemic administration of the P08908 receptor agonist 8-OH-2-(di-n-propylamino)-tetralin ( 8-OH-DPAT ; 0.3 mg/kg , s.c. ) was used to explore the effects of activation of P08908 receptors on expression of mRNA coding for P08908 receptor , tryptophan hydroxylase ( P17752 ) and galanin in the ascending raphe nuclei . 8-OH-DPAT increased the hybridization signal of the P08908 receptor by 105 % in the dorsal raphe nucleus ( P33681 ) 30 min after the injection . No effects were seen at the later time points ( 2-8 h ) . In the median raphe nucleus ( B8 ) and the B9 cell group in the medial lemniscus , 8-OH-DPAT induced a marked decrease in labeling 30 min after injection . At 8 h following 8-OH-DPAT injection , the effect had shifted to an increase in P08908 receptor labeling by 68 % in the B8 area . Importantly 8-OH-DPAT had no significant effects on the expression of mRNA coding for P17752 and galanin . The results suggest an important and differential mechanism for the regulation of P08908 receptor mRNA levels in the dorsal and median raphe nuclei . This regulation may be of importance for the differential control of the activity of the ascending 5-HT neurons , and hence for mood regulation . The results also indicate a dissociation between the effects mediated by P08908 receptor functions and those regulating the coexisting peptide galanin in the dorsal raphe .