MH_dev_93

The low-potency , voltage-dependent Q12809 REA blocker propafenone - - molecular determinants and drug trapping . The molecular determinants of high-affinity human ether-a-go-go-related gene ( Q12809 REA ) potassium channel blockade by methanesulfonanilides include two aromatic residues ( Phe 656 and Tyr 652 ) on the inner helices ( S6 ) and residues on the pore helices that face into the inner cavity , but determinants for lower-affinity Q12809 REA blockers may be different . In this study , alanine-substituted Q12809 REA channel mutants of inner cavity residues were expressed in Xenopus laevis oocytes and were used to characterize the Q12809 REA channel binding site of the antiarrhythmic propafenone . DB01182 MEN ' s blockade of Q12809 REA was strongly dependent on residue Phe 656 but was insensitive or weakly sensitive to mutation of Tyr 652 , Thr 623 , Ser 624 , Val 625 , Gly 648 , or Val 659 and did not require functional inactivation . Homology models of Q12809 REA based on KcsA and MthK crystal structures , representing the closed and open forms of the channel , respectively , suggest propafenone is trapped in the inner cavity and is unable to interact exclusively with Phe 656 in the closed state ( whereas exclusive interactions between propafenone and Phe 656 are found in the open-channel model ) . These findings are supported by very slow recovery of wild-type Q12809 REA channels from block at - 120 mV , but extremely rapid recovery of D540K channels that reopen at this potential . The experiments and modeling suggest that the open-state propafenone binding-site may be formed by the Phe 656 residues alone . The binding site for propafenone ( which may involve pi-stacking interactions with two or more Phe 656 side-chains ) is either perturbed or becomes less accessible because of closed-channel gating . This provides further evidence for the existence of gating-induced changes in the spatial location of Phe 656 side chains .

DB00171 - sensitive potassium channels ( K ( DB00171 ) ) in retina : a key role for delayed ischemic tolerance . The objectives of the present study were to determine the localization of K ( DB00171 ) channels in normal retina and to evaluate their potential roles in ischemic preconditioning ( IPC ) in a rat model of ischemia induced by increased intraocular pressure ( IOP ) . Brown Norway rats were subjected to sublethal 3 - , lethal 20 - and 40 - min ischemia and the functional recovery was evaluated using electroretinography . The time interval between ischemic insults ranged from 1 to 72 h . The effects of K ( DB00171 ) channel blockade on IPC protection were studied by treatment with 0.01 % glipizide . IPC was mimicked by injection of K ( DB00171 ) channel openers of 0.01 % ( - ) cromakalim or 0.01 % P1060 72 h before 20 - min ischemia . Co-expression of K ( DB00171 ) channel subunits Kir 6.2 / Q09428 REA was observed in the retinal pigment epithelium , inner segments of photoreceptors , outer plexiform and ganglion cell layers and at the border of the inner nuclear layer . In contrast to a 20 - or 40 - min ischemia , a 3 - min ischemia induced no alteration of the electroretinogram ( ERG ) and constituted the preconditioning stimulus . An ischemic challenge of 40 min in preconditioned rats induced impairment of retinal function . However , animals preconditioned 24 , 48 and 72 h before 20 - min ischemia had a significant improvement of the ERG . ( - ) Cromakalim and P1060 mimicked the effect of IPC . DB01067 MEN significantly suppressed the protective effects of preconditioning . In conclusion , activation of K ( DB00171 ) channels plays an important role in the mechanism of preconditioning by enhancing the resistance of the retina against a severe ischemic insult .

Serotonergic modulation of the acoustic startle response in rats during preweaning development . The involvement of serotonin ( 5 - HT ) in modulating the acoustic startle response ( ASR ) is well established in adult rats , but 5 - HT involvement during the preweaning period , when 5 - HT neurons undergo extensive development , has not previously been described . Three 5 - HT receptor subtypes are reported to modulate the ASR in adult rats : P08908 REA and 5 - HT2 receptor agonists facilitate the ASR , whereas P28222 REA agonists decrease the response . In the present study , the effects of 5 - HT agonists and generalized 5 - HT depletion on the ASR were studied in preweanling animals , using independent groups of Long-Evans rats tested on postnatal day ( P01160 REA ) 13 , 17 and 21 . 8 - Hydroxy - 2 - ( di-n-propylamino ) tetralin ( 8OHDPAT , 62-1000 micrograms / kg ) , a P08908 REA receptor agonist , and 5 - methoxy-N , N-dimethyl tryptamine ( MeODMT , 2-4 mg / kg ) , a nonselective 5 - HT agonist , had no effect on P01160 REA 13 and then increased the ASR on P01160 REA 17 and 21 . The 5 - HT2 receptor antagonists cyproheptadine ( 5 mg / kg ) and ketanserin ( 5 mg / kg ) blocked the effect of MeODMT at both ages , providing some evidence that MeODMT increased the ASR through 5 - HT2 receptors . 1 - ( m-Chlorophenyl ) piperazine ( mCPP , 1-5 mg / kg ) , a P28222 REA agonist , had no effect on ASR amplitude on P01160 REA 13 or 17 and then produced a dose-related decrease in the response on P01160 REA 21 . Generalized depletion of 5 - HT by 80-90 % in whole-brain and spinal cord , using p-chlorophenylalanine ( PCPA , 300 mg / kg 24 hr prior to testing ) , did not alter ASR amplitude at any age . ( ABSTRACT TRUNCATED AT 250 WORDS )

Differences in transcript levels of ABC transporters between pancreatic adenocarcinoma and nonneoplastic tissues . OBJECTIVES : The aim of this study was to evaluate transcript levels of all 49 human DB00171 - binding cassette transporters ( ABCs ) in one of the most drug-resistant cancers , namely , the pancreatic ductal adenocarcinoma ( PDAC ) . Association of ABCs levels with clinical-pathologic characteristics and P01116 REA mutation status was followed as well . METHODS : Tumors and adjacent nonneoplastic tissues were obtained from 32 histologically verified PDAC patients . The transcript profile of ABCs was assessed using quantitative real-time polymerase chain reaction with a relative standard curve . P01116 REA mutations in exon 2 were assessed by high-resolution melting analysis and sequencing . RESULTS : Most ABCs were deregulated in PDAC and 10 ABCs were associated with clinical-pathologic characteristics . P01116 REA mutations did not change the global expression profile of ABCs . CONCLUSIONS : The expression of ABC transporters was significantly deregulated in PDAC tumors when compared to nonmalignant tissues . The observed up-regulation of P21439 REA , O95342 REA , P33527 REA , O15438 REA , O15440 REA , Q5T3U5 REA , and Q9UNQ0 in tumors may contribute to the generally poor treatment response of PDAC . The up-regulation of O95477 REA , Q8IZY2 , and P45844 REA implicates a serious impairment of cellular cholesterol homeostasis in PDAC . On the other hand , the observed down-regulation of Q99758 REA , O95255 REA , P13569 REA , and Q09428 REA suggests a possible role of stem cells in the development and progression of PDAC .

A DNA hypermethylation profile reveals new potential biomarkers for prostate cancer diagnosis and prognosis . BACKGROUND : DNA hypermethylation has emerged as a novel molecular biomarker for the evaluation of prostate cancer diagnosis and prognosis . Defining the specific gene hypermethylation profile for prostate cancer could involve groups of genes that specifically discriminate patients with indolent and aggressive tumors . METHODS : Genome-wide methylation analysis was performed on 83 tumor and 10 normal prostate samples using the GoldenGate Methylation Cancer Panel I ( Illumina , Inc . ) . All clinical stages of disease were considered . RESULTS : We found 41 genes hypermethylated in more than 20 % of the tumors analyzed ( P < 0.01 ) . Of these , we newly identified P28161 REA and P01210 REA as being genes that are hypermethylated in prostate cancer and that were simultaneously methylated in 40.9 % of the tumors analyzed . We also identified panels of genes that are more frequently methylated in tumor samples with clinico-pathological indicators of poor prognosis : a high Gleason score , elevated Ki - 67 , and advanced disease . Of these , we found simultaneous hypermethylation of P13569 REA and P28222 REA to be common in patients with a high Gleason score and high Ki - 67 levels ; this might indicate the population at higher risk of therapeutic failure . The DNA hypermethylation profile was associated with cancer-specific mortality ( log-rank test , P = 0.007 ) and biochemical recurrence-free survival ( log-rank test , P = 0.0008 ) . CONCLUSIONS : Our findings strongly indicate that epigenetic silencing of P28161 REA and P01210 REA is a common event in prostate cancer that could be used as a molecular marker for prostate cancer diagnosis . In addition , simultaneous P28222 REA and P13569 REA hypermethylation could help discriminate aggressive from indolent prostate tumors .

Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 REA , Q12809 REA , Q14524 REA , P22460 REA , Q9UK17 , P15382 REA , 2 , 5 , P63252 REA , P35498 REA - 3B , P01160 REA , and P36382 REA from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = . 0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF .

A review of the use of frovatriptan in the treatment of menstrually related migraine . Menstrual migraine ( MM ) is a highly prevalent condition associated with considerable disability . Migraine attacks occur exclusively around the menstrual period in approximately 10 % of women with migraine , that is , pure menstrual migraine , while at least 50 % of them also experience migraine at other times of the month , that is , menstrually related migraine ( MRM ) . The therapeutic approach to patients with MRM is based on treatment of the attack , or prophylactic strategies . Triptans are recommended as first-line treatments for moderate to severe migraine attacks , including MM . DB00998 SUB is one of the newest triptans . Its high affinity for P28222 REA / 1D receptors and long half-life contribute to its distinctive clinical effect , characterized by a more sustained and prolonged effect than other triptans . Indeed , frovatriptan proved to be effective in treating the acute attack , but was particularly effective in the short-term preventive therapy of MM . In addition , frovatriptan is one of the safest triptans , with the lowest risk of treatment-emergent adverse events . Following extensive evidence from randomized pharmacological trials , frovatriptan has now gained a grade A recommendation from the guidelines for short-term prophylaxis of MM . Recent post-hoc analyses of direct comparative trials also suggest that frovatriptan might have an important role in the acute treatment of MRM . In these studies , frovatriptan showed pain relief and pain-free rates similar to those of zolmitriptan , rizatriptan , and almotriptan , but with significantly lower recurrence rates . More well-designed , randomized , prospective studies , specifically enrolling women with MM , will be needed in the near future to confirm the efficacy of frovatriptan in this migraine subtype .

DB00998 SUB : a selective type 1B / 1D serotonin receptor agonist for the treatment of migraine headache . DB00998 SUB belongs to an innovative family of compounds aimed at breaking through the long-standing barrier of migraine headache understanding and treatment . While a typology of headaches has been recognized for some time , and a number of therapies have been introduced for reduction of headache pain and duration , the causes of migraine remain a subject of debate . Those prone to attacks continue to endure them and suffer the related symptoms such as nausea and disorientation . DB00998 SUB , like all the triptans , acts by inducing vasoconstriction of the meningeal arteries . It has been shown in pharmacological tests to act selectively as a potent agonist of serotonin P28222 REA / 1D receptors . DB00998 SUB has been well tolerated in humans and efficacious in reducing headache pain and duration in clinical trials , which have also indicated that dose adjustments for age or gender are not necessary for the drug . Patients have found the use of frovatriptan acceptable over the long-term , and overall a low-incidence of adverse effects has been reported . Though not a prophylactic , frovatriptan has demonstrated the potential to significantly improve the therapeutic approaches to the treatment of migraine .

DB00998 SUB . black triangle DB00998 SUB , a new serotonin receptor agonist developed for the acute treatment of migraine , has high affinity for serotonin P28222 REA and P28221 REA receptor subtypes and is a potent stimulator of contraction in human basilar arteries . black triangle A long terminal elimination half-life ( approximately 26 hours ) is a distinctive pharmacokinetic feature of frovatriptan which appears to be independent of dose , age , gender and renal function . black triangle A single oral dose of frovatriptan 2.5 mg was effective in the acute treatment of migraine providing meaningful relief within 2 hours to approximately twice as many recipients as placebo in clinical trials . black triangle Consistent relief of migraine symptoms was achieved in patients who treated a number of consecutive attacks with frovatriptan and the incidence of 24 - hour migraine recurrence was reduced . black triangle DB00998 SUB was well tolerated in clinical trials , with the overall incidence of adverse events occurring with frovatriptan 2.5 mg only slightly higher than that reported with placebo . Mild to moderate fatigue , nausea and paraesthesia were the most commonly reported drug-related adverse events .

DB00998 SUB Vernalis . Vanguard ( now Vernalis ) has developed frovatriptan , a selective P28222 REA / 1D partial agonist licensed from GlaxoSmithKline as a potential treatment for migraine [ 188478 ] , [ 194382 ] , [ 377863 ] .

Structure functional expression and spatial distribution of a cloned cDNA encoding a rat P28221 REA - like receptor . Using polymerase chain reaction ( PCR ) a complementary DNA ( cDNA ) encoding a 5 - hydroxytryptamine ( 5 - HT ) receptor was isolated from rat forebrain . The amplified cDNA specifies an open reading frame of 374 amino acids comprising seven putative transmembrane regions . Expression of the cloned cDNA in human embryonic kidney cells ( P29320 REA 293 ) was used to establish the pharmacological profile of the encoded receptor polypeptide . Membranes containing the cloned receptor showed high affinity binding of [ 3H ] - 5 - HT . Competition binding experiments with a variety of serotonin receptor ligands displayed a rank order of affinities corresponding to a P28221 REA subtype : 5 - CT > 5 - HT = metergoline > CGS 12066 > methysergide > sumatriptan > mianserin = ( - ) alpha-Me - 5 - HT = yohimbine > 8 - OH-DPAT > or = rauwolscine > spiperone > DOI > propranolol > or = 2 - Me - 5 - HT > or = ICS 205930 . Ketanserin and ritanserin displaced [ 3H ] - 5 - HT-binding in a biphasic manner . In situ hybridization revealed highest expression of the corresponding mRNA in the pyramidal layer of the olfactory tubercle and the nucleus caudatus and accumbens .

DB00175 MEN - induced proangiogenic effects depend upon extracellular P09038 REA . The P04035 REA inhibitors ( statins ) have been shown to exert several protective effects on the vasculature that are unrelated to changes in the cholesterol profile , and to induce angiogenesis . The proangiogenic effect exerted by statins has been attributed to the activation of the PI3K / Akt pathway in endothelial cells ; however , it is unclear how statins activate this pathway . DB00175 MEN - mediated activation of Akt and MAPK occurs rapidly ( within 10 min . ) and at low doses ( 10 nM ) . Here , we hypothesized that P09038 REA contributes to the proangiogenic effect of statins . We found that pravastatin , a hydrophilic statin , induced phosphorylation of the FGF receptor ( FGFR ) in human umbilical vein endothelial cells . SU5402 , an inhibitor of FGFR , abolished pravastatin-induced PI3K / Akt and MAPK activity . Likewise , anti - P09038 REA function-blocking antibodies inhibited Akt and MAPK activity . Moreover , depletion of extracellular P09038 REA by heparin prevented pravastatin-induced phosphorylation of Akt and MAPK . Treatment with P09038 REA antibody inhibited pravastatin-enhanced endothelial cell proliferation , migration and tube formation . These observations indicate that pravastatin exerts proangiogenic effects in endothelial cells depending upon the extracellular P09038 REA .

Estrogen regulation of uterine genes in vivo detected by complementary DNA array . INTRODUCTION : In the present study , our aim was to identify differentially expressed genes involved in estrogen actions at the endometrium level in rats . METHODS : Thirty adult rats were ovariectomized four days prior to drug administration for 48 days . Rats were divided in 2 groups : I , control and II , conjugated equine estrogens ( CCE ) . Total RNA was isolated from uterus , and differential expression was analyzed by array technology and RT-PCR . RESULTS : A total of 32 candidate genes were shown to be upregulated or downregulated in groups I or II . Among them , differential expression was already confirmed by RT-PCR for P24593 REA , P28222 REA , c-kit , and P15692 REA , genes whose expression was up regulated during CCE therapy , and casein kinase II and serine kinase expression was the same level in both groups . CONCLUSION : We have demonstrated that cDNA array represents a powerful approach to identify key molecules in the estrogens therapy . A number of the candidates reported here should provide new markers that may contribute to the detection of target estrogen receptor . This information may also aid the development of new approaches to therapeutic intervention .

Correcting human mitochondrial mutations with targeted RNA import . Mutations in the human mitochondrial genome are implicated in neuromuscular diseases , metabolic defects , and aging . An efficient and simple mechanism for neutralizing deleterious mitochondrial DNA ( mtDNA ) alterations has unfortunately remained elusive . Here , we report that a 20 - ribonucleotide stem-loop sequence from the H1 RNA , the RNA component of the human RNase P enzyme , appended to a nonimported RNA directs the import of the resultant RNA fusion transcript into human mitochondria . The methodology is effective for both noncoding RNAs , such as tRNAs , and mRNAs . The RNA import component , polynucleotide phosphorylase ( Q8TCS8 ) , facilitates transfer of this hybrid RNA into the mitochondrial matrix . In addition , nucleus-encoded mRNAs for mitochondrial proteins , such as the mRNA of human mitochondrial ribosomal protein P28222 REA ( O15235 REA ) , contain regulatory sequences in their 3 ' - untranslated region ( UTR ) that confers localization to the mitochondrial outer membrane , which is postulated to aid in protein translocation after translation . We show that for some mitochondrial-encoded transcripts , such as P35354 REA , a 3 ' - UTR localization sequence is not required for mRNA import , whereas for corrective mitochondrial-encoded tRNAs , appending the 3 ' - UTR localization sequence was essential for efficient fusion-transcript translocation into mitochondria . In vivo , functional defects in mitochondrial RNA ( mtRNA ) translation and cell respiration were reversed in two human disease lines . Thus , this study indicates that a wide range of RNAs can be targeted to mitochondria by appending a targeting sequence that interacts with Q8TCS8 , with or without a mitochondrial localization sequence , providing an exciting , general approach for overcoming mitochondrial genetic disorders .

Shared and unique genetic contributions to attention deficit / hyperactivity disorder and substance use disorders : a pilot study of six candidate genes . The shared genetic basis of attention deficit / hyperactivity disorder ( ADHD ) and substance use disorders ( SUDs ) was explored by investigating the association of candidate risk factors in neurotransmitter genes with both disorders . One hundred seven methadone maintenance treatment patients , 36 having an ADHD diagnosis , 176 adult patients with ADHD without SUDs , and 500 healthy controls were genotyped for variants in the P21917 REA ( exon 3 VNTR ) , P21918 REA ( upstream VNTR ) , P28222 REA ( rs6296 ) , P09172 REA ( rs2519152 ) , P21964 REA ( rs4680 ; Val 158Met ) , and P35372 REA ( rs1799971 ; 118A > G ) genes . Association with disease was tested using logistic regression models . This pilot study was adequately powered to detect larger genetic effects ( OR ≥ 2 ) of risk alleles with a low frequency . Compared to controls , ADHD patients ( with and without SUDs ) showed significantly increased frequency of the P09172 REA ( rs2519152 : OR 1.73 ; CI 1.15- 2.59 ; P= 0.008 ) and the P35372 REA risk genotypes ( rs1799971 : OR 1.71 ; CI 1.17- 2.50 ; P= 0.006 ) . The P09172 REA risk genotype was associated with ADHD diagnosis , with the association strongest in the pure ADHD group . The P35372 REA risk genotype increased the risk for the combined ADHD and SUD phenotype . The present study strengthens the evidence for a shared genetic basis for ADHD and addiction . The association of P35372 REA with the ADHD and SUD combination could help to explain the contradictory results of previous studies . The power limitations of the study restrict the significance of these findings : replication in larger samples is warranted .

Fluorescence energy transfer analysis of calmodulin-peptide complexes . The interactions between calmodulin and the tryptophan residues of synthetic peptides corresponding to the calmodulin binding domains of skeletal muscle myosin light-chain kinase and the plasma membrane calcium pump were examined . The single tryptophan residue contained in each peptide became relatively immobilized and inaccessible to iodide ion upon binding to calmodulin , indicating that the indole side chain was inserted into a hydrophobic cleft in the surface of calmodulin . Fluorescence energy transfer from peptidyl tryptophan residues to an AEDANS moiety attached to cysteine - 26 of spinach calmodulin was measured . Included in these analyses was a tryptophan-containing peptide analog of the calmodulin binding domain of neuromodulin . These data indicated that the indole ring of each peptide inserted 32-35 A away from cysteine - 26 and may therefore interact with the carboxyl-terminal lobe of P62158 in its " bent " conformation [ Persechini & Kretsinger ( 1988a ) J . Cardiovasc . Pharmacol . 12 ( Suppl 5 ) , S1 - P28222 REA ; Ikura et al . ( 1992 ) Science 256 , 632-638 ; Vorherr et al . ( 1992 ) Eur . J . Biochem . 204 , 931-937 ] . The interchange of tryptophan - 3 and phenylalanine - 21 of the calcium pump peptide increased the efficiency of energy transfer to the AEDANS-moiety approximately 12 - fold , reducing the calculated distance to 20 A . These data suggest that phenylalanine - 21 of the calcium pump peptide interacts with the hydrophobic cleft in the amino-terminal lobe of P62158 .

Meta-analysis of oral DB00669 therapy for migraine : number needed to treat and relative cost to achieve relief within 2 hours . OBJECTIVE : To determine the cost-effectiveness of the P28222 REA / 1D agonists , or triptans , in the acute treatment of migraine . METHODS : To determine the cost-effectiveness of the triptans , a meta-analysis was conducted of the efficacy data from 27 oral DB00669 trials , using the endpoint of " pain-free " status within 2 hours after initial dosing as the indicator of efficacy . Efficacy data were used to determine the number needed to treat ( Q13423 REA ) to achieve pain-free status in 1 patient within 2 hours postdose and then applied the per-dose costs for each DB00669 to the Q13423 REA values . RESULTS : Rizatriptan 10 mg and almotriptan 12.5 mg were the most cost-effective of the triptans , costing $ 48.34 and $ 48.57 US dollars , respectively , to achieve pain-free status in 1 patient within 2 hours postdose . DB00998 SUB 2.5 mg was the most costly , with a cost-effective ratio of $ 162.49 US dollars . All other triptans fell between these extremes : zolmitriptan 5 mg ( $ 65.18 US dollars ) , sumatriptan 100 mg ( $ 70.83 US dollars ) , sumatriptan 50 mg ( $ 75.67 US dollars ) , zolmitriptan 2.5 mg ( $ 78.74 US dollars ) , and naratriptan 2.5 mg ( $ 141.43 US dollars ) , in decreasing order of cost-effectiveness . CONCLUSION : Using an Q13423 REA analysis , the least-costly drugs to achieve migraine cure within 2 hours are rizatriptan 10 mg and almotriptan 12.5 mg . From a population health perspective , the lower acquisition cost of almotriptan 12.5 mg allows for effective treatment of more patients than rizatriptan 10 mg for no additional medication cost .

Opposed effects of lithium on the MEK - P29323 REA pathway in neural cells : inhibition in astrocytes and stimulation in neurons by GSK 3 independent mechanisms . DB01356 MEN is widely used in the treatment of bipolar disorder , but despite its proven therapeutic efficacy , the molecular mechanisms of action are not fully understood . The present study was undertaken to explore lithium effects of the MEK / P29323 REA cascade of protein kinases in astrocytes and neurons . In asynchronously proliferating rat cortical astrocytes , lithium decreased time - and dose-dependently the phosphorylation of MEK and P29323 REA , with 1 mM concentrations achieving 60 and 50 % inhibition of P29323 REA and MEK , respectively , after a 7 - day exposure . DB01356 MEN also inhibited [ 3H ] thymidine incorporation into DNA and induced a G2 / M cell cycle arrest . In serum-deprived , quiescent astrocytes , pre-exposure to lithium resulted in the inhibition of cell cycle re-entry as stimulated by the mitogen endothelin - 1 : under this experimental setting , lithium did not affect the rapid , peak phosphorylation of MEK taking place after 3-5 min , but was effective in inhibiting the long-term , sustained phosphorylation of MEK . DB01356 MEN inhibition of the astrocyte MEK / P29323 REA pathway was independent of inositol depletion . Further , compound SB216763 inhibited Tau phosphorylation at Ser 396 and stabilized cytosolic beta-catenin , consistent with the inhibition of glycogen synthase kinase - 3 beta ( P49841 REA ) , but failed to reproduce lithium effects on MEK and P29323 REA phosphorylation and cell cycle arrest . In cerebellar granule neurons , millimolar concentrations of lithium enhanced MEK and P29323 REA phosphorylation in a concentration-dependent manner , again through an inositol and P49841 REA independent mechanism . These opposing effects in astrocytes and neurons make lithium treatment a promising strategy to favour neural repair and reduce reactive gliosis after traumatic injury .

The efficacy and tolerability of frovatriptan and dexketoprofen for the treatment of acute migraine attacks . DB00998 SUB is a DB00669 characterized by a high affinity for P28222 REA / 1D receptors and a long half-life contributing to a more sustained and prolonged action than other triptans . DB09214 is a nonsteroidal anti-inflammatory drug with a relatively short half-life and rapid onset of action , blocking the action of cyclo-oxygenase , which is involved in prostaglandins ' production , thus reducing inflammation and pain . Both drugs have been successfully employed as monotherapies for the treatment of acute migraine attacks . The combination of these two drugs ( frovatriptan 2.5 mg plus dexketoprofen 25 or 37.5 mg ) has been tested in migraine sufferers , showing a rapid and good initial efficacy , with 2 - h pain free rates of 51 % , and a high persistence in the 48 - h following the onset of pain : recurrence occurred in only 29 % of attacks and sustained pain free rates were 43 % at 24 - and 33 % at 48 - h .

Signalling pathways involved in retinal endothelial cell proliferation induced by advanced glycation end products : inhibitory effect of gliclazide . AIM : We have previously demonstrated that advanced glycation end products ( AGEs ) stimulate bovine retinal endothelial cell ( BREC ) proliferation through induction of vascular endothelial growth factor ( P15692 REA ) production by these cells . We have also shown that gliclazide , a sulfonylurea which decreases oxidative stress , inhibits this effect . The aim of the present study was to characterize the signalling pathways involved in P51606 REA - induced BREC proliferation and P15692 REA production and mediating the inhibitory effect of gliclazide on these biological events . METHODS : BRECs were treated or not treated with AGEs in the presence or absence of gliclazide , antioxidants , protein kinase C ( PKC ) , mitogen-activated protein kinase ( MAPK ) or nuclear factor-kappaB ( NF-kappaB ) inhibitors . BREC proliferation was assessed by measuring [ 3H ] - thymidine incorporation into DNA . Activation of PKC , MAPK and NF-kappaB signal transduction pathways and determination of P15692 REA expression were assessed by Western blot analysis using specific antibodies . MAPK activity was also determined by an in vitro kinase assay . RESULTS : Treatment of BRECs with AGEs significantly increased cell proliferation and P15692 REA expression . AGEs induced P05771 REA translocation , extracellular signal-regulated protein kinase 1/2 and NF-kappaB activation in these cells . Pharmacological inhibition of these signalling pathways abolished P51606 REA effects on cell proliferation and P15692 REA expression . Exposure of BRECs to gliclazide or antioxidants such as vitamin E or N-acetyl-l-cysteine resulted in a significant decrease in P51606 REA - induced activation of PKC - , MAPK - and NF-kappaB-signalling pathways . CONCLUSIONS : Our results demonstrate the involvement of PKC , MAPK and NF-kappaB in P51606 REA - induced BREC proliferation and P15692 REA expression . DB01120 MEN inhibits BREC proliferation by interfering with these intracellular signal transduction pathways .

DB00998 SUB , a P28222 REA / 1D receptor agonist for migraine . DB00998 SUB is one of the most recent serotonin receptor agonists to receive FDA , approved labelling for use in the acute management of migraine with or without aura in adults . The mechanism of action of frovatriptan is thought to be similar to that of a serotonin agonist . However , frovatriptan has distinctive pharmacokinetic and pharmacologic properties , chiefly , a high affinity for serotonin receptors 1B and 1D and a long elimination half-life ; frovatriptan was shown to be more selective for cerebral than coronary arteries , a property which makes frovatriptan more favourable in patients at risk of coronary artery disease . Additionally , frovatriptan has a half-life of approximately 25 h , substantially longer than that of any other agent within its class . This property makes frovatriptan suitable for patients who typically suffer migraines of long duration and / or those who suffer migraine recurrence . The efficacy of frovatriptan in the treatment of acute migraine was demonstrated in five double-blind , randomised , placebo-controlled trials . At 2h , headache response rates for frovatriptan 2.5 mg ranged from 38 to 40 % compared to 22-35 % for placebo . Headache recurrence for frovatriptan 2.5 mg at 24h ranged from 9 to 14 % compared with 18 % in placebo subjects . DB00998 SUB has no clinically significant pharmacokinetic interactions with drugs used for migraine prophylaxis or with commonly prescribed medications . Adverse effects of frovatriptan including dizziness , paresthesia , dry mouth , fatigue and flushing were generally mild and well tolerated . Given the fact that patient response to serotonin agonists is individualised , and selecting an effective agent may involve trial and error , frovatriptan is a welcome alternative in the acute management of migraine .

P35372 REA and P20813 REA gene variants as risk factors in methadone-related deaths . DB00333 MEN is a medication valued for its effectiveness in the treatment of heroin addiction ; however , many fatal poisonings associated with its use have been reported over the years . We have examined the association between P20813 REA and micro-opioid receptor ( P35372 REA ) gene variations and apparent susceptibility to methadone poisoning . Genomic DNA was extracted from postmortem whole blood of 40 individuals whose deaths were attributed to methadone poisoning . The presence of P20813 REA * 4 , * 9 , and * 6 alleles and the P35372 REA A118G variant was determined by SNP genotyping . P20813 REA * 4 , * 9 , and * 6 alleles were found to be associated with higher postmortem methadone concentrations in blood ( P < or = 0.05 ) . P35372 REA A118G was also associated with higher postmortem methadone concentrations in blood but not to a level of statistical significance ( P = 0.39 ) . In these methadone-related deaths , P35372 REA 118GA was associated with higher postmortem benzodiazepine concentrations ( P = 0.04 ) , a finding not associated with morphine-related deaths . The risk of a methadone-related fatality during treatment may be evaluated in part by screening for P20813 REA * 6 and A118G .

Effects of external calcium on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . DB01373 is a known signalling molecule in eukaryotic cells and plays a central role in the regulation of many cellular processes . In the following study , we report on the effect of external calcium treatments on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . We observed that the intracellular calcium content of P . bainier 229-7 mycelia was increased in response to exposure to high external Ca ( 2 + ) concentrations . Both ginsenoside Rd biotransformation and β-glucosidase activity were both found to be dependent on the external calcium concentration . At an optimal Ca ( 2 + ) concentration of 45 mM , maximal ginsenoside Rd bioconversion rate of 92.44 % was observed and maximal β-glucosidase activity of 0.1778 U was reached in a 72 - h biotransformation . The Ca ( 2 + ) channel blocker Verapamil blocked the trans-membrane influx of calcium and decreased ginsenoside Rd biotransformatiom . In addition , β-glucosidase activity and ginsenoside Rd content decreased by 36.0 and 29.2 % respectively after a 72 - h incubation in the presence of 0.05 mM P62158 ( P62158 ) antagonist DB00850 MENMAX DB00850 MEN . These results suggest that both Ca ( 2 + ) channels and P62158 are involved in ginsenoside Rd biotransformation via regulation of β-glucosidase activity . This is the first report regarding the effects of calcium signal transduction on biotransformation and enzyme activity in fungi .

Rationalizing cyclooxygenase ( P36551 REA ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase - 2 ( P35354 REA ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 REA inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 REA inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 REA isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 REA inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 MEN ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10 - fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 REA inhibitors can be compared .

A new cell culture-based assay quantifies vitamin K 2,3- epoxide reductase complex subunit 1 function and reveals warfarin resistance phenotypes not shown by the dithiothreitol-driven Q9BQB6 assay . BACKGROUND : DB00682 MEN directly inhibits the vitamin K 2,3- epoxide reductase complex subunit 1 ( Q9BQB6 ) enzyme to effect anticoagulation . Q9BQB6 function has historically been assessed in vitro using a dithiothreitol ( DTT ) - driven vitamin K 2,3- epoxide reductase ( Q9BQB6 ) assay . DB00682 MEN inhibits wild-type Q9BQB6 function by the DTT - Q9BQB6 assay . However , Q9BQB6 variants with warfarin resistance-associated missense mutations often show low Q9BQB6 activities and warfarin sensitivity instead of resistance . OBJECTIVES : A cell culture-based , indirect Q9BQB6 assay was developed and characterized that accurately reports warfarin sensitivity or resistance for wild-type and variant Q9BQB6 proteins . METHODS : Human coagulation factor ( F ) IX and Q9BQB6 variants were coexpressed in P29320 REA 293T cells under standardized conditions at various warfarin concentrations . Secreted FIX activity served as surrogate marker to report wild-type and variant Q9BQB6 inhibition by warfarin . RESULTS AND CONCLUSIONS : DB00682 MEN dose-response curves fit to the secreted FIX activity data for coexpressed hVKORC 1 wild-type , Val 29Leu , Val 45Ala and Leu 128Arg variants . The corresponding calculated IC50 values were 24.7 , 136.4 , 152.0 and 1226.4 nm , respectively . Basal activities in the absence of warfarin for all Q9BQB6 variants were similar to that of wild-type Q9BQB6 . Ranked IC50 values from the cell culture-based assay accurately reflect elevated warfarin dosages for patients with Q9BQB6 missense mutation-associated warfarin resistance .

Activation of Akt 1 by human 5 - hydroxytryptamine ( serotonin ) 1B receptors is sensitive to inhibitors of MEK . Akt 1 / protein kinase B and the mitogen-activated protein ( Q96HU1 ) kinases extracellular signal-regulated kinase 1 ( P27361 REA ) and P28482 REA have been shown to promote cell survival in a cell-specific manner . Since many receptors activate both pathways , inhibitors are commonly used to study the relative role of each pathway . In the present study , we examined the effects of PD098059 and U0126 , two structurally dissimilar inhibitors of Q96HU1 kinase kinase ( Q02750 REA / 2 ) , on the activation of P29323 REA and Akt stimulated by human 5 - hydroxytryptamine ( 1B ) ( serotonin ) ( P28222 REA ) receptors . Surprisingly , pathways for activation of both P29323 REA and Akt were found to be sensitive to the two MEK inhibitors at concentrations commonly used to selectively inhibit the activation of P29323 REA . Both compounds caused complete inhibition of phosphorylation of P29323 REA and a maximal 60 % inhibition of P28222 REA receptor-mediated phosphorylation of Akt . Inhibition of Akt activation required almost complete inhibition of P29323 REA . Transfection with cDNA for activated forms of Q02750 REA / 2 caused increased phosphorylation of P29323 REA but not of Akt , demonstrating that independent activation of MEK / P29323 REA was insufficient for activation of Akt . Therefore , it is not clear whether inhibition of activation of Akt resulted from selective inhibition of MEK or from additional actions on other unidentified common pathways . Nevertheless , our findings that PD098059 and U0126 inhibit activation of Akt at commonly used concentrations demonstrate that in at least some systems , these compounds inhibit activation of both P29323 REA and Akt , and can not be used to discern the relative roles of each pathway in mediating cellular responses .

Pharmacokinetic evaluation of frovatriptan . INTRODUCTION : Migraine is the most common painful neurological disorder , affecting 13 % of the general population . Triptans represent a powerful pharmacological tool in acute migraine treatment , however , a significant portion of treated patients can not have access to this class due to possible adverse affects . Today , a total of seven DB00669 molecules are available , representing a commonly prescribed migraine treatment . Although there is a need of extensive use of triptans , only 25 % of migraine patients are using triptans . AREAS COVERED : This review includes triptans and evidence for the use of frovatriptan . A systematic approach is used to discuss the pharmacodynamic and pharmacokinetic aspects of frovatriptan , considering the emerging data on the clinical efficacy of frovatriptan in the treatment of migraine and cluster headaches . The data were obtained by searching the following key words in MEDLINE : pharmacokinetic , pharmacodynamic , triptans , frovatriptan , migraine , menstrual migraine , relatively to the period 1988 - 2011 . EXPERT OPINION : DB00998 SUB has been developed in order to improve safety and efficacy of triptans . It shows a favorable tolerability and efficacy profile , limited to 24/48- h headache recurrence , when compared with other triptans . Preclinical data suggest that the pharmacokinetic profile of frovatriptan may differ from other available triptans . In fact , among triptans , frovatriptan showed the highest potency at the P28222 REA receptor ( 8.2 ) and the longer half-life ( 26 h ) . These parameters determine the clinical properties of frovatriptan ; in particular the lowest rate of headache recurrence in comparison with other triptans .

Constitutive G ( i2 ) - dependent activation of adenylyl cyclase type II by the P08908 REA receptor . Inhibition by anxiolytic partial agonists . The P08908 REA receptor is implicated in depression and anxiety . This receptor couples to G ( i ) proteins to inhibit adenylyl cyclase ( AC ) activity but can stimulate AC in tissues ( e . g . hippocampus ) that express ACII . The role of ACII in receptor-mediated stimulation of DB02527 formation was examined in P29320 REA - 293 cells transfected with the P08908 REA receptor , which mediated inhibition of basal and G ( s ) - induced DB02527 formation in the absence of ACII . In cells cotransfected with P08908 REA receptor and ACII plasmids , P08908 REA agonists induced a 1 . 5 - fold increase in DB02527 level . Cotransfection of P08908 REA receptor , ACII , and Galpha ( i2 ) , but not Galpha ( i1 ) , Galpha ( i3 ) , or Galpha ( o ) , resulted in an agonist-independent 6 - fold increase in the basal DB02527 level , suggesting that G ( i2 ) preferentially coupled the receptor to ACII . The P28222 REA receptor also constitutively activated ACII . Constitutive activity of the P08908 REA receptor was blocked by pertussis toxin and the Gbetagamma antagonist , betaCT , suggesting an important role for Gbetagamma-mediated activation of ACII . The DB00156 - 149 --> Ala mutation in the second intracellular domain of the P08908 REA receptor disrupted Gbetagamma-selective activation of ACII . Spontaneous P08908 REA receptor activity was partially attenuated by P08908 REA receptor partial agonists with anxiolytic activity ( e . g . buspirone and flesinoxan ) but was not altered by full agonists or antagonists . Thus , anxiolytic activity may involve inhibition of spontaneous P08908 REA receptor activity .

Differential functional activity of 5 - hydroxytryptamine receptor ligands and beta adrenergic receptor antagonists at 5 - hydroxytryptamine 1B receptor sites in Chinese hamster lung fibroblasts and opossum renal epithelial cells . Functional activity of 5 - hydroxytryptamine ( 5 - HT ) receptor ligands and beta adrenergic receptor antagonists was studied at P28222 REA receptor sites in Chinese hamster lung ( CHL ) fibroblasts by measuring two cellular responses : inhibition of forskolin-stimulated cyclic AMP formation and potentiation of basic fibroblast growth ( BFGF ) induced mitogenesis . A good correlation was found between the potency of agonists to inhibit forskolin-induced cyclic AMP formation and their potency to potentiate P09038 REA - induced thymidine incorporation in CHL fibroblasts . Potent agonist activity was measured with 5 - methoxy -3,1 , 2,3 , 6 - tetrahydro - 4 - pyidinyl - 1H - indole ( RU 24,969 ) , 5 - carboxamidotryptamine ( 5 - CT ) , 3 - ( 1,2 , 5,6 ) - tetrahydro - 4 - pyridyl - 5 - pyrrolo ( 3,2- b ) pyril - 5 - one ( CP 93,129 ) and 5 - HT , whereas sumatriptan displayed weak agonist activity at concentrations different from its binding affinity for P28222 REA binding sites . In contrast to the observed P28222 REA receptor-mediated agonist activity in opossum kidney cells for metergoline and the beta adrenergic receptor antagonists : cyanopindolol , 4 - ( 3 - tert-butyl-amino - 2 - hydroxypropoxy ) - indole - 2 carbonic acid isopropyl ester ( SDZ 21,009 ) , isamoltane , ( - ) - propranolol and ( - ) - pindolol , antagonist activity at P28222 REA receptor sites was yielded in CHL fibroblasts in accordance with the reported observations at rat brain P28222 REA receptors . Methiothepin was the only compound that antagonized both the opossum kidney cell and CHL fibroblast P28222 REA receptor-mediated responses although the antagonist effect was more pronounced in CHL fibroblasts . ( ABSTRACT TRUNCATED AT 250 WORDS )

Involvement of P28222 REA receptors in DB00669 - induced contractile responses in guinea-pig isolated iliac artery . Using a series of triptans we characterized in vitro the 5 - hydroxytryptamine ( 5 - HT ) receptor that mediates the contraction in guinea-pig iliac arteries moderately precontracted by prostaglandin F2alpha ( PGF 2alpha ) . Additionally , we investigated by reverse-transcriptase polymerase chain reaction ( RT-PCR ) which DB00669 - sensitive receptor is present in this tissue . DB00998 SUB , zolmitriptan , rizatriptan , naratriptan , sumatriptan , and almotriptan contracted guinea-pig iliac arteries with pD2 values of 7.52+ / -0.04 , 6.72+ / -0.03 , 6.38+ / -0.06 , 6.22+ / -0.05 , 5.86+ / -0.05 and 5.26+ / -0.04 respectively . For comparison , the pD2 values for 5 - HT and 5 - carboxamidotryptamine ( 5 - CT ) were 7.52+ / -0.02 and 7.55+ / -0.03 respectively . In contrast to all other triptans tested , the concentration-response curve for eletriptan was biphasic ( first phase : 0.01- 3 microM , pD2 approximately 6.6 ; second phase : > or = 10 microM ) . Contractions to 5 - HT , 5 - CT , frovatriptan , zolmitriptan , rizatriptan , naratriptan , sumatriptan , almotriptan , and eletriptan ( first phase ) were antagonized by the P28222 REA / 1D receptor antagonist GR127935 ( 10 nM ) and the P28222 REA receptor antagonist SB216641 ( 10 nM ) . RT-PCR studies in guinea-pig iliac arteries showed a strong signal for the P28222 REA receptor while expression of P28221 REA and P30939 REA receptors was not detected in any sample . The present results demonstrate that DB00669 - induced contraction in guinea-pig iliac arteries is mediated by the P28222 REA receptor . The guinea-pig iliac artery may be used as a convenient in vitro model to study the ( cardio ) vascular side-effect potential of anti-migraine drugs of the DB00669 family .