interacts_with DB02546?
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 . tert-Butylcarbamate-containing histone deacetylase inhibitors : apoptosis induction , cytodifferentiation , and antiproliferative activities in cancer cells . Herein we report novel pyrrole- and benzene-based hydroxamates ( 8 , 10 ) and 2'-aminoanilides ( 9 , 11 ) bearing the tert-butylcarbamate group at the CAP moiety as histone deacetylase ( HDAC ) inhibitors . Compounds 8 b and 10 c selectively inhibited Q9UBN7 at the nanomolar level , whereas the other hydroxamates effected an increase in acetyl-α-tubulin levels in human acute myeloid leukemia U937 cells . In the same cell line , compounds 8 b and 10 c elicited 18.4 and 21.4 % apoptosis , respectively ( DB02546 : 16.9 % ) , and the pyrrole anilide 9 c displayed the highest cytodifferentiating effect ( 90.9 % ) . In tests against a wide range of various cancer cell lines to determine its antiproliferative effects , compound 10 c exhibited growth inhibition from sub-micromolar ( neuroblastoma LAN-5 and SH-SY5Y cells , chronic myeloid leukemia K562 cells ) to low-micromolar ( lung H1299 and A549 , colon HCT116 and HT29 cancer cells ) concentrations . In HT29 cells , 10 c increased histone H3 acetylation , and decreased the colony-forming potential of the cancer cells by up to 60 % . 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 . Selective inhibition of histone deacetylase 6 ( Q9UBN7 ) induces DNA damage and sensitizes transformed cells to anticancer agents . Q9UBN7 ( Q9UBN7 ) is structurally and functionally unique among the 11 human zinc-dependent histone deacetylases . Here we show that chemical inhibition with the Q9UBN7 -selective inhibitor tubacin significantly enhances cell death induced by the topoisomerase II inhibitors etoposide and doxorubicin and the pan-HDAC inhibitor DB02546 ( vorinostat ) in transformed cells ( LNCaP , MCF-7 ) , an effect not observed in normal cells ( human foreskin fibroblast cells ) . The inactive analogue of tubacin , nil-tubacin , does not sensitize transformed cells to these anticancer agents . Further , we show that down-regulation of Q9UBN7 expression by shRNA in LNCaP cells enhances cell death induced by etoposide , doxorubicin , and DB02546 . Tubacin in combination with DB02546 or etoposide is more potent than either drug alone in activating the intrinsic apoptotic pathway in transformed cells , as evidenced by an increase in PARP cleavage and partial inhibition of this effect by the pan-caspase inhibitor Z-VAD-fmk . Q9UBN7 inhibition with tubacin induces the accumulation of γ P16104 , an early marker of DNA double-strand breaks . Tubacin enhances DNA damage induced by etoposide or DB02546 as indicated by increased accumulation of γ P16104 and activation of the checkpoint kinase Chk2 . Tubacin induces the expression of P35638 ( P35638 / P35638 ) , a transcription factor up-regulated in response to cellular stress . P35638 induction is further increased when tubacin is combined with DB02546 . These findings point to mechanisms by which Q9UBN7 -selective inhibition can enhance the efficacy of certain anti-cancer agents in transformed cells . 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 . 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 ' . Effects of DB00482 and DB00744 on liver metastasis and lipidperoxidation in pancreatic cancer in Syrian hamsters . Selective inhibition of eicosanoid synthesis is thought to have effects on carcinogenesis in lung and colon cancer . However , it is still unknown whether pancreatic cancer might also be influenced . Therefore we evaluated the impact of selective cyclooxygenase-2 inhibitor DB00482 and selective P09917 inhibitor DB00744 on liver metastasis in a solid model of pancreatic adenocarcinoma in Syrian hamster . In week 33 , the animals were sacrificed and incidence of pancreatic carcinomas and number and size of liver metastases were determined . Activities of antioxidative enzymes ( GSHPX/SOD ) and concentrations of products of lipidperoxidation were measured in liver metastases and non-metastatic hepatic tissue . The incidence ( 54.5 vs. 100 % ) , number ( 3.17 +/- 0.98 vs. 6.75 +/- 0.71 ) and size ( 2.67 +/- 1.97 vs. 11.75 +/- 1.98 mm2 ) of liver metastases were decreased by combined therapy of DB00744 and DB00482 ( P < 0.05 ) . Furthermore , activities of GSHPX ( [ 73.77 +/- 5.67 ] *10(5) vs. [ 15.49 +/- 4.02 ] *10(5) U/mg prot. ; P < 0.05 ) and SOD ( 474.92 +/- 108.8 vs. 127.89 +/- 38.75 U/mg prot. ; P < 0.05 ) were increased , while lipidperoxidation ( 0.31 +/- 0.08 nmol/mg prot. vs. 1.54 +/- 0.55 nmol/mg prot. ; P < 0.05 ) was decreased by combination therapy , in non-metastatic hepatic tissue . Moreover , combined therapy increased lipidperoxidation in liver metastases ( 0.47 +/- 0.09 vs. 1.95 +/- 0.12 nmol/mg prot. ; P < 0.05 ) . Thus , a combination of DB00482 and DB00744 might be a new concept to decrease tumour growth in liver metastases in advanced pancreatic cancer . 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 . Primary resistance of P51681 -tropic HIV-1 to maraviroc can not be predicted by the V3 sequence . OBJECTIVES : Resistance of HIV-1 to P51681 antagonists can occur without coreceptor switching by mutations in envelope glycoproteins that enable virus entry using the inhibitor-bound form of P51681 . We investigated whether mutations in the V3 region of HIV-1 from subjects naive to maraviroc could be associated with primary resistance to this drug . METHODS : The frequency of P51681 -tropic HIV-1 subtype B isolates harbouring putative V3 maraviroc resistance mutations was assessed among the HIV tropism database of Toulouse University Hospital , France . Phenotypic assessment of maraviroc susceptibility was performed for 14 isolates representative of the main mutation patterns and 14 controls . V3 mutations were reversed or introduced by site-directed mutagenesis . RESULTS : Ninety-three of 951 ( 9.8 % ) isolates harboured V3 mutations assumed to be associated with maraviroc resistance . DB04835 completely blocked virus entry for all but 1 of the 14 isolates harbouring V3 mutations [ IC50 8.6 nM ; 95 % CI ( 6.6-47.4 ) ] , as in the 14 control isolates [ IC50 13.4 nM ; 95 % CI ( 7.7-50.3 ) ] ( P = 0.24 ) . Primary resistance to maraviroc , with a plateau in entry inhibition , was found in one isolate ( harbouring a 20F/21I genotype ) . Site-directed mutagenesis showed that V3 mutations are necessary but not sufficient to induce maraviroc resistance . CONCLUSIONS : The impact of V3 mutations depended on the env context in which they occurred . Simple assessment of the V3 genotype thus can not accurately predict maraviroc resistance . Rather , phenotypic assessment of virus particles expressing the envelope glycoprotein as a whole is required . This approach revealed that primary resistance of P51681 -tropic HIV-1 subtype B isolates to maraviroc seems uncommon . Interactions of alpha- , beta- , and theta-defensins with influenza A virus and surfactant protein D . We have reported that the alpha-defensins human neutrophil peptides ( HNP ) -1 and HNP-2 neutralize and aggregate influenza A virus ( IAV ) and promote uptake of IAV by neutrophils . These alpha-defensins were also shown to bind to surfactant protein ( SP ) -D and reduce its antiviral activity . In this study , we examined retrocyclin (RC)1 and RC2 , humanized versions of the antiviral theta-defensins found in the leukocytes of certain nonhuman primates . Q9HC52 was just as effective as P59665 -3 in neutralizing IAV , and RC2 and RC101 ( an analog of Q9HC52 ) were more effective . In contrast , human beta-defensins ( HBDs ) showed less neutralizing activity . Human defensins 5 and 6 ( mainly produced by intestinal Paneth cells ) had viral neutralizing activity similar to P59665 -3 . Like P59665 -3 , RCs induced viral aggregation and promoted the uptake of IAV by neutrophils . We used surface plasmon resonance to evaluate binding of defensins to P35247 . HBDs , Q9UBN7 , and P12838 bound minimally to P35247 . P59665 -3 and RCs bound P35247 with high affinity ; however , unlike P59665 and HNP-2 , RCs did not inhibit P35247 antiviral activity . HBDs also did not inhibit antiviral activity of P35247 . Given their strong neutralizing activity and compatibility with P35247 , RCs may provide attractive prototypes for designing therapeutics that can prevent or treat respiratory infections caused by IAV . Q9NQT5 , the N-terminal domain of p35 , protects against Q00535 /p25-induced neurotoxicity . P12004 -dependent kinase 5( Q00535 ) in complex with its activator , p35 ( protein of 35 kDa ) , is essential for early neurodevelopment in mammals . However , endogenous cleavage of p35 to p25 is associated with neuron death and neurodegenerative disease . Here we show that a peptide ( Q9NQT5 ' ) encoding the N-terminal domain of p35 protects against Q00535 /p25-induced toxicity in neurons . Q9NQT5 ' also prevented the death of neurons treated with the neurotoxin , 1-methyl-4-phenylpyridinium ( P25189 (+) ) , which induces conversion of endogenous p35 to p25 , and Parkinson disease ( PD ) -like symptoms in animals . P25189 (+) induces Q00535 /p25-dependent phosphorylation of peroxiredoxin 2 ( Prx2 ) , resulting in inhibition of its peroxireductase activity and accumulation of reactive oxygen species ( ROS ) . We found that Q9NQT5 ' expression inhibited both Prx2 phosphorylation and ROS accumulation in neurons . In addition , Q9NQT5 ' inhibited the p25-induced appearance of antigen of the Ki67 antibody ( Ki67 ) and phosphohistone P16104 ( γ P16104 ) , classic markers of cell cycle activity and DNA double-strand breakage , respectively , associated with neuron death . Our results suggest that Q9NQT5 ( protein of 10 kDa ) is a unique prosurvival domain in p35 , essential for normal Q00535 /p35 function in neurons . Loss of the Q9NQT5 domain results in Q00535 /p25 toxicity and neurodegeneration in vivo . Expression of DNA-dependent protein kinase holoenzyme upon induction of lymphocyte differentiation and V(D)J recombination . Murine preB lymphocytes grow in tissue culture in the presence of stromal cells and interleukin 7 ( P13232 ) , and can be induced to differentiate to surface-immunoglobulin-positive B cells in vitro by withdrawal of P13232 . Upon differentiation , proliferation ceases , and upregulation of Rag-1 and Rag-2 expression , and induction of V(D)J immunoglobulin-gene rearrangements occur . DNA-dependent protein kinase ( DNA-PK ) is required for effective V(D)J recombination and repair of DNA double-strand breaks . The holoenzyme comprises a catalytic subunit ( P78527 ) and the Ku heterodimer ( P12956 / P13010 ) . We have analyzed expression of P12956 , P13010 and P78527 upon induction of differentiation in preB cells derived from wild-type , severe combined immunodeficiency ( SCID ) and Rag-2-/- mice . Protein levels of P13010 and P12956 moderately decrease after induction in all three cell types . A distinct polypeptide that crossreacts with anti-Ku Ig appears in the cytoplasm of wild-type and Rag-2-/- cells , but not of SCID cells . In mouse preB cells , P12956 and P13010 are present in the nuclei and cytoplasm before and after onset of differentiation . In vivo , P12956 is predominantly expressed in V(D)J-recombination-active , early-preB and P01730 -/CD8- thymocyte cell populations . Upon differentiation , protein levels of P78527 are unaltered . DNA-PK activity , which is not detectable in SCID cells , increases in wild-type and Rag-2-/- cells more than twofold shortly after induction of differentiation , then falls back to about 50 % of starting levels . DB02546 , an HDAC inhibitor attenuates epidermoid squamous cell carcinoma growth by dampening P42345 signaling pathway in a human xenograft murine model . Histone deacetylase ( HDAC ) inhibitors are potent anticancer agents and show efficacy against various human neoplasms . DB02546 is a potent HDAC inhibitor and has shown potential to inhibit growth of human xenograft tumors . However , its effect on the growth of skin neoplasm remains undefined . In this study , we show that vorinostat ( 2 μM ) reduced expression of Q13547 , 2 , 3 , and 7 in epidermoid carcinoma A431 cells . Consistently , it increased acetylation of histone H3 and p53 . DB02546 ( 100mg/kg body weight , IP ) treatment reduced human xenograft tumor growth in highly immunosuppressed nu/nu mice . Histologically , the vorinostat-treated tumor showed features of well-differentiation with large necrotic areas . Based on proliferating cell nuclear antigen ( P12004 ) staining and expression of cyclins D1 , D2 , E , and A , vorinostat seems to impair proliferation by down-regulating the expression of these proteins . However , it also induced apoptosis . The mechanism by which vorinostat blocks proliferation and makes tumor cells prone to apoptosis , involved inhibition of P42345 signaling which was accompanied by reduction in cell survival AKT and extracellular-signal regulated kinase ( P29323 ) signaling pathways . Our data provide a novel mechanism-based therapeutic intervention for cutaneous squamous cell carcinoma ( SCC ) . DB02546 may be utilized to cure skin neoplasms in organ transplant recipient ( OTR ) . These patients have high morbidity and surgical removal of these lesions which frequently develop in these patients , is difficult . Differential gene signatures in rat mammary tumors induced by DMBA and those induced by fractionated gamma radiation . The aim of this work was to identify specific genes involved in rat mammary tumors induced by dimethylbenz(a)anthracene ( DMBA ) or radiation . More TUNEL- and P12004 -positive cells were present in mammary tumors induced by radiation than in tumors induced by DMBA , whereas DNA damage responses like p53 accumulation and histone P16104 phosphorylation were higher in DMBA-induced tumors , even though the pathology was similar in both types of tumors . cDNA microarray and real-time RT-PCR analysis of radiation- or DMBA-induced tumor tissues , revealed that stanniocalcin 2 ( Stc2 ) , interferon regulatory factor 1 ( Irf1 ) , interleukin 18 binding protein ( Il18bp ) , and chloride channel calcium activated 3 ( Clca3 ) were expressed in both , and that arachidonate P09917 activating protein 1 ( Alox5ap ) and cathepsin S ( Ctss ) were expressed only in radiation-induced tumors . No DMBA-specific gene signatures were found . Soft agar growth assays were carried out to identify the carcinogenic features of these specific genes . Cells stably transfected with Alox5ap , Ctss , Stc2 , Irf1 , Il18bp and Clca3 showed morphological changes compared to controls . These findings indicate different gene alterations in carcinogen- or radiation-induced mammary tumors with similar pathological stages . 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 . Calorie restriction promotes mammalian cell survival by inducing the Q96EB6 deacetylase . A major cause of aging is thought to result from the cumulative effects of cell loss over time . In yeast , caloric restriction ( CR ) delays aging by activating the Sir2 deacetylase . Here we show that expression of mammalian Sir2 ( Q96EB6 ) is induced in CR rats as well as in human cells that are treated with serum from these animals . P01308 and insulin-like growth factor 1 ( DB01277 ) attenuated this response . Q96EB6 deacetylates the DNA repair factor P12956 , causing it to sequester the proapoptotic factor Bax away from mitochondria , thereby inhibiting stress-induced apoptotic cell death . Thus , CR could extend life-span by inducing Q96EB6 expression and promoting the long-term survival of irreplaceable cells . 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 . 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 . 3,3',4 ' , 5'-Tetramethoxychalcone inhibits human oral cancer cell proliferation and migration via p53-mediated mitochondrial-dependent apoptosis . BACKGROUND/AIM : The current study aimed to identify an attractive target against human oral squamous cell carcinoma ( OSCC ) . MATERIALS AND METHODS : The effect of 3,3',4',5'-tetramethoxychalcone ( P46977 ) on OSCC cell proliferation , cell-cycle phase distribution , expression of markers of cell apoptosis , and cell migration were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay , flow cytometry , western blot , and transwell migration assay , respectively . RESULTS : Experimental results revealed that P46977 inhibited the OSCC cell proliferation ( fifty percent inhibitory concentrations range=1.0-4.5 μM ) by inducing G2/M phase arrest of the cell cycle . P46977 caused DNA double-strand breaks , and enhanced expression of caspase-3 and -9 , poly ( ADP-ribose ) polymerase , cytochrome c , calpain-1 and -2 , phosphorylation of histone P16104 , phosphorylation of checkpoint kinases 2 , p53 , P10415 -antagonist/killer and P10415 -associated × protein , while reducing the mitochondrial membrane potential , and expression of B-cell lymphoma-2 . In addition , P46977 reduced the migration potential of OSCC cells by attenuating the C-C chemokine ligand 5/ P51681 axis . CONCLUSION : These data indicate that P46977 may be considered an interesting target for further development of chemotherapeutic agents against oral cancer . A case study of acenocoumarol sensitivity and genotype-phenotype discordancy explained by combinations of polymorphisms in Q9BQB6 and P11712 . To determine the cause of a genotype-phenotype discordancy for acenocoumarol sensitivity . Methods A patient , highly sensitive to acenocoumarol , and previously determined to carry only a single P11712 *3 allele , was genotyped for additional functionally defective alleles in the P11712 and Q9BQB6 genes . Family members were also analyzed to trace the pedigree . Results The acenocoumarol-sensitive patient was found to possess , in addition to P11712 *3 allele , a P11712 *11 allele and the Q9BQB6 AA diplotype which were all traced back through the parental lines . Conclusions DB01418 sensitivity in this subject is the consequence of inheritance of multiple functionally defective alleles in both the P11712 and Q9BQB6 genes . The study provides additional data in support of diminished P11712 activity due to the presence of the rare *11 allele . In vitro effects and in vivo efficacy of a novel cyclooxygenase-2 inhibitor in cats with lipopolysaccharide-induced pyrexia . OBJECTIVE : To determine cyclooxygenase ( P36551 ) -2 selectivity , pharmacokinetic properties , and in vivo efficacy of firocoxib ( Q9NTI2 ,785,713 ) in cats . ANIMALS : 5 healthy male and 14 healthy female domestic shorthair cats . PROCEDURE : Selectivity of firocoxib for inhibiting P35354 was determined by comparing the potency for inhibiting P23219 with that of P35354 in feline blood . Pharmacokinetic properties were determined after i.v. ( 2 mg/kg ) and oral ( 3 mg/kg ) administration in male cats . In vivo efficacy was evaluated in female cats with lipopolysaccharide ( LPS ) -induced pyrexia with administration of firocoxib 1 or 14 hours before LPS challenge . RESULTS : Blood concentrations resulting in 50 % inhibition of P23219 and P35354 activity in vitro were 75 +/- 2 microM and 0.13 +/- 0.03 microM , respectively , and selectivity for inhibiting P35354 relative to P23219 was 58 . DB09217 had moderate to high oral bioavailability ( 54 % to 70 % ) , low plasma clearance ( 4.7 to 5.8 mL/min/kg ) , and an elimination half-life of 8.7 to 12.2 hours . DB09217 at doses from 0.75 to 3 mg/kg was efficacious in attenuating fever when administered to cats 1 or 14 hours before LPS challenge . CONCLUSIONS AND CLINICAL RELEVANCE : DB09217 is a potent P35354 inhibitor and is the only selective P35354 inhibitor described for use in cats to date . It is effective in attenuating febrile responses in cats when administered 14 hours before LPS challenge , suggesting it would be suitable for once-a-day dosing . Because selective P35354 inhibitors have an improved therapeutic index relative to nonselective nonsteroidal anti-inflammatory drugs in humans , firocoxib has the potential to be a safe , effective anti-inflammatory agent for cats . The anti-tumor effect of HDAC inhibition in a human pancreas cancer model is significantly improved by the simultaneous inhibition of cyclooxygenase 2 . Pancreatic ductal adenocarcinoma is the fourth leading cause of cancer death worldwide , with no satisfactory treatment to date . In this study , we tested whether the combined inhibition of cyclooxygenase-2 ( P35354 ) and class I histone deacetylase ( HDAC ) may results in a better control of pancreatic ductal adenocarcinoma . The impact of the concomitant HDAC and P35354 inhibition on cell growth , apoptosis and cell cycle was assessed first in vitro on human pancreas BxPC-3 , PANC-1 or CFPAC-1 cells treated with chemical inhibitors ( DB02546 , MS-275 and celecoxib ) or Q13547 /2/3/7 siRNA . To test the potential antitumoral activity of this combination in vivo , we have developed and characterized , a refined chick chorioallantoic membrane tumor model that histologically and proteomically mimics human pancreatic ductal adenocarcinoma . The combination of Q13547 /3 and P35354 inhibition significantly impaired proliferation of BxPC-3 cells in vitro and stalled entirely the BxPC-3 cells tumor growth onto the chorioallantoic membrane in vivo . The combination was more effective than either drug used alone . Consistently , we showed that both Q13547 and O15379 inhibition induced the expression of P35354 via the NF-kB pathway . Our data demonstrate , for the first time in a Pancreatic Ductal Adenocarcinoma ( PDAC ) model , a significant action of HDAC and P35354 inhibitors on cancer cell growth , which sets the basis for the development of potentially effective new combinatory therapies for pancreatic ductal adenocarcinoma patients .