MH_dev_241

An acetylcholinesterase inhibitor , eserine , induces long-term depression at P07451 REA - P00915 REA 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 REA ( P22303 REA ) 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 REA - P00915 REA synapses ; weak pharmacological activation using carbachol ( CCh ) facilitates potentiation , whereas strong agonism induces muscarinic long-term depression ( mLTD ) via an P29323 REA - dependent mechanism . Here , we tested the prediction that accumulation of extracellular ACh via inhibition of P22303 REA is sufficient to induce LTD at P07451 REA - P00915 REA synapses in hippocampal slices from adult rats . Although P22303 REA inhibition with eserine induces LTD , it unexpectedly does not share properties with mLTD induced by CCh , as reported previously . DB00981 MEN - 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 REA inhibition causes a prolonged decrease in presynaptic glutamate release at P07451 REA - P00915 REA 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 SUB : 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-SY 5Y cells , chronic myeloid leukemia K562 cells ) to low-micromolar ( lung H1299 and A549 , colon HCT 116 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 MEN 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 REA ) 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 REA ) is absent , Q16822 REA message and protein were detected in P01308 REA - 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 REA - 1 832/13 cells and islets measured substantial contribution of Q16822 REA to the synthesis of PEP . As high as 30 % of PEP in P01308 REA - 1 832/13 cells and 41 % of PEP in rat islets came from Q16822 REA . The contribution of Q16822 REA to overall PEP synthesis more than tripled with glucose stimulation . Silencing the Q16822 REA 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 REA 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 SUB ( 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 SUB . Tubacin in combination with DB02546 SUB 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 REA , an early marker of DNA double-strand breaks . Tubacin enhances DNA damage induced by etoposide or DB02546 SUB as indicated by increased accumulation of γ P16104 REA and activation of the checkpoint kinase Chk 2 . Tubacin induces the expression of P35638 REA ( P35638 REA / P35638 REA ) , a transcription factor up-regulated in response to cellular stress . P35638 REA induction is further increased when tubacin is combined with DB02546 SUB . 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 REA / 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 MEN , the most active paullone , was demonstrated to act by competing with DB00171 for binding to GSK - 3beta . DB04014 MEN inhibits the phosphorylation of tau in vivo at sites which are typically phosphorylated by GSK - 3beta in Alzheimer ' s disease . DB04014 MEN also inhibits the Q00535 REA / 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 REA / 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 REA that regulates FLIP protein stability by inhibiting its polyubiquitination . Furthermore , we found that the histone deacetylase ( HDAC ) inhibitor DB02546 SUB ( DB02546 SUB ) enhances the acetylation of P12956 REA , thereby disrupting the FLIP / P12956 REA complex and triggering FLIP polyubiquitination and degradation by the proteasome . Using in vitro and in vivo colorectal cancer models , we further demonstrated that DB02546 SUB - induced apoptosis is dependant on FLIP downregulation and caspase 8 activation . In addition , an Q9UBN7 - specific inhibitor Tubacin recapitulated the effects of DB02546 SUB , suggesting that Q9UBN7 is a key regulator of P12956 REA 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 MEN 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 REA inhibitor DB00744 MEN 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 MEN 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 MEN 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 REA ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 REA ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 REA ) at 3 and 12 months . P10632 REA * 4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 REA levels ( P= 0.003 and P= 0.007 ) and were 2.2- and 2.5- fold more likely to develop P42126 REA and CND ( P= 0.039 and P= 0.041 ) , respectively . P34913 REA 55Arg , P51589 REA * 7 , P10632 REA * 1B , and P10632 REA g . 36785A allele carriers had lower EET and DHET P04141 REA levels . P10632 REA g . 25369T and P10632 REA g . 36755A allele carriers had higher EET levels . Patients with P10632 REA * 2C and P34913 REA 404del variants had worse long-term outcomes while those with P34913 REA 287Gln , P51589 REA * 7 , and P11712 REA 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 REA - tropic HIV - 1 to maraviroc can not be predicted by the V3 sequence . OBJECTIVES : Resistance of HIV - 1 to P51681 REA antagonists can occur without coreceptor switching by mutations in envelope glycoproteins that enable virus entry using the inhibitor-bound form of P51681 REA . 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 REA - 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 MEN 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 REA - 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 REA - 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 REA - 3 . Like P59665 REA - 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 REA . HBDs , Q9UBN7 , and P12838 REA bound minimally to P35247 REA . P59665 REA - 3 and RCs bound P35247 REA with high affinity ; however , unlike P59665 REA and HNP - 2 , RCs did not inhibit P35247 REA antiviral activity . HBDs also did not inhibit antiviral activity of P35247 REA . Given their strong neutralizing activity and compatibility with P35247 REA , 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 REA / p25 - induced neurotoxicity . P12004 REA - dependent kinase 5 ( Q00535 REA ) 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 REA / p25 - induced toxicity in neurons . Q9NQT5 ' also prevented the death of neurons treated with the neurotoxin , 1 - methyl - 4 - phenylpyridinium ( P25189 REA ( + ) ) , which induces conversion of endogenous p35 to p25 , and Parkinson disease ( PD ) - like symptoms in animals . P25189 REA ( + ) induces Q00535 REA / p25 - dependent phosphorylation of peroxiredoxin 2 ( Prx 2 ) , resulting in inhibition of its peroxireductase activity and accumulation of reactive oxygen species ( ROS ) . We found that Q9NQT5 ' expression inhibited both Prx 2 phosphorylation and ROS accumulation in neurons . In addition , Q9NQT5 ' inhibited the p25 - induced appearance of antigen of the Ki67 antibody ( Ki67 ) and phosphohistone P16104 REA ( γ P16104 REA ) , 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 REA / p35 function in neurons . Loss of the Q9NQT5 domain results in Q00535 REA / 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 REA ) , and can be induced to differentiate to surface-immunoglobulin-positive B cells in vitro by withdrawal of P13232 REA . 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 REA ) and the Ku heterodimer ( P12956 REA / P13010 REA ) . We have analyzed expression of P12956 REA , P13010 REA and P78527 REA upon induction of differentiation in preB cells derived from wild-type , severe combined immunodeficiency ( SCID ) and Rag - 2 - / - mice . Protein levels of P13010 REA and P12956 REA 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 REA and P13010 REA are present in the nuclei and cytoplasm before and after onset of differentiation . In vivo , P12956 REA is predominantly expressed in V ( D ) J-recombination-active , early-preB and P01730 REA - / CD8 - thymocyte cell populations . Upon differentiation , protein levels of P78527 REA 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 SUB , an HDAC inhibitor attenuates epidermoid squamous cell carcinoma growth by dampening P42345 REA signaling pathway in a human xenograft murine model . Histone deacetylase ( HDAC ) inhibitors are potent anticancer agents and show efficacy against various human neoplasms . DB02546 SUB 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 REA , 2 , 3 , and 7 in epidermoid carcinoma A431 cells . Consistently , it increased acetylation of histone H3 and p53 . DB02546 SUB ( 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 REA ) 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 REA signaling which was accompanied by reduction in cell survival AKT and extracellular-signal regulated kinase ( P29323 REA ) signaling pathways . Our data provide a novel mechanism-based therapeutic intervention for cutaneous squamous cell carcinoma ( SCC ) . DB02546 SUB 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 REA - 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 REA 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 ( Stc 2 ) , interferon regulatory factor 1 ( Irf 1 ) , interleukin 18 binding protein ( Il18bp ) , and chloride channel calcium activated 3 ( Clca 3 ) were expressed in both , and that arachidonate P09917 REA activating protein 1 ( Alox 5ap ) 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 Alox 5ap , Ctss , Stc 2 , Irf 1 , Il18bp and Clca 3 showed morphological changes compared to controls . These findings indicate different gene alterations in carcinogen - or radiation-induced mammary tumors with similar pathological stages .

P43490 REA / P43490 REA / visfatin and cancer . P43490 REA / P43490 REA / 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 REA and stem cell factor . A number of cancers have increased expression of P43490 REA / P43490 REA / visfatin , which regulates a variety of different signaling pathways such as PI3K / Akt , P27361 REA / 2 and P40763 REA . FK866 / APO 866 and CHS 828 / DB05217 MEN are two known inhibitors of P43490 REA / P43490 REA / 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 Sir 2 deacetylase . Here we show that expression of mammalian Sir 2 ( Q96EB6 ) is induced in CR rats as well as in human cells that are treated with serum from these animals . P01308 REA and insulin-like growth factor 1 ( DB01277 ) attenuated this response . Q96EB6 deacetylates the DNA repair factor P12956 REA , 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 REA ) potassium channel . OBJECTIVE : To evaluate the effects of azimilide and ambasilide on the biophysical properties of the human-ether-a-go-go-related ( Q12809 REA ) channel . METHODS : Q12809 REA was stably transfected into Chinese hamster ovary ( CHO - P04264 REA ) cells and currents were measured using a whole cell , voltage-clamp technique . RESULTS : DB04957 MEN 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 REA 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 REA 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 REA tagged with enhanced green fluorescent protein . The Ku protein is a complex of two subunits , P12956 REA and P13010 REA , 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 REA and P13010 REA 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 REA tagged with the green fluorescent protein ( GFP ) color variants in P13010 REA - deficient cells , i . e . , xrs - 6 derived from CHO - P04264 REA . Although P12956 REA , as well as P13010 REA , was undetectable in xrs - 6 cells , it was seen in these transformants at a level similar to the level of CHO - P04264 REA . Furthermore , etoposide - and radiosensitive phenotype of xrs - 6 cells were corrected by an introduction of the tagged P13010 REA . Moreover , the tagged P13010 REA 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 REA in the Ku-dependent DSB repair . Therefore , these transformants might be useful not only in the analysis of P13010 REA 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 REA ) 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 REA 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 REA 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 REA , phosphorylation of checkpoint kinases 2 , p53 , P10415 REA - antagonist / killer and P10415 REA - associated × protein , while reducing the mitochondrial membrane potential , and expression of B-cell lymphoma - 2 . In addition , P46977 REA reduced the migration potential of OSCC cells by attenuating the C-C chemokine ligand 5 / P51681 REA axis . CONCLUSION : These data indicate that P46977 REA 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 REA . 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 REA * 3 allele , was genotyped for additional functionally defective alleles in the P11712 REA 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 REA * 3 allele , a P11712 REA * 11 allele and the Q9BQB6 AA diplotype which were all traced back through the parental lines . Conclusions DB01418 MENMAX DB01418 MEN sensitivity in this subject is the consequence of inheritance of multiple functionally defective alleles in both the P11712 REA and Q9BQB6 genes . The study provides additional data in support of diminished P11712 REA 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 REA ) - 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 REA was determined by comparing the potency for inhibiting P23219 REA with that of P35354 REA 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 REA and P35354 REA activity in vitro were 75 + / - 2 microM and 0.13 + / - 0.03 microM , respectively , and selectivity for inhibiting P35354 REA relative to P23219 REA was 58 . DB09217 MEN 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 MEN 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 MEN is a potent P35354 REA inhibitor and is the only selective P35354 REA 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 REA 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 REA ) and class I histone deacetylase ( HDAC ) may results in a better control of pancreatic ductal adenocarcinoma . The impact of the concomitant HDAC and P35354 REA 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 SUB , MS - 275 and celecoxib ) or Q13547 REA / 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 REA / 3 and P35354 REA 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 REA and O15379 REA inhibition induced the expression of P35354 REA 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 REA inhibitors on cancer cell growth , which sets the basis for the development of potentially effective new combinatory therapies for pancreatic ductal adenocarcinoma patients .