interacts_with DB01012?
DB01012 HCl prevents development of parathyroid gland hyperplasia and reverses established parathyroid gland hyperplasia in a rodent model of CKD . BACKGROUND : Secondary hyperparathyroidism ( sHPT ) represents an adaptive response to progressively impaired control of calcium , phosphorus and vitamin D in chronic kidney disease ( CKD ) . It is characterized by parathyroid hyperplasia and excessive synthesis and secretion of parathyroid hormone ( PTH ) . Parathyroid hyperplasia in uremic rats can be prevented by calcium-sensing receptor ( P41180 ) activation with the calcimimetic cinacalcet ( Sensipar®/Mimpara® ) ; however , it is unknown , how long the effects of cinacalcet persist after withdrawal of treatment or if cinacalcet is efficacious in uremic rats with established sHPT . METHODS : We sought to determine the effect of cinacalcet discontinuation in uremic rats and whether cinacalcet was capable of influencing parathyroid hyperplasia in animals with established sHPT . RESULTS : Discontinuation of cinacalcet resulted in reversal of the beneficial effects on serum PTH and parathyroid hyperplasia . In rats with established sHPT , cinacalcet decreased serum PTH and mediated regression of parathyroid hyperplasia . The cinacalcet-mediated decrease in parathyroid gland size was accompanied by increased expression of the cyclin-dependent kinase inhibitor P38936 . Prevention of cellular proliferation with cinacalcet occurred despite increased serum phosphorus and decreased serum calcium . CONCLUSIONS : The animal data provided suggest established parathyroid hyperplasia can be reversed by modulating P41180 activity with cinacalcet and that continued treatment may be necessary to maintain reductions in PTH . Epithelial calcium-sensing receptor activation by eosinophil granule protein analog stimulates collagen matrix contraction . BACKGROUND : Eosinophils reside in normal gastrointestinal tracts and increase during disease states . Receptors for eosinophil-derived granule proteins ( EDGPs ) have not been identified , but highly cationic molecules , similar to eosinophil proteins , bind extracellular calcium-sensing receptors ( CaSRs ) . We hypothesized that stimulation of CaSRs by eosinophil proteins activates epithelial cells . METHODS : Caco2 intestinal epithelial cells , AML14.3D10 eosinophils , wild-type ( WT ) human embryonic kidney 293 ( HEK293 ) cells not expressing CaSRs ( P29320 -WT ) , and P41180 -transfected HEK293 cells ( P29320 - P41180 ) were stimulated with an eosinophil protein analog poly-L-arginine ( PA ) and phosphorylated extracellular signal-regulated kinase (pERK)1 and pERK2 were measured . Functional activation was measured with collagen lattice contraction assays . RESULTS : Coculture of Caco2 cells with AML14.3D10 eosinophils augmented lattice contraction as compared with lattices containing Caco2 cells alone . PA stimulation of Caco2 lattices augmented contraction . P29320 - P41180 stimulation with PA or Ca(2+) resulted in greater pERK activation than that of stimulated P29320 -WT cells . PA stimulated greater P29320 - P41180 lattice contraction than unstimulated lattices . Contraction of PA-stimulated and PA-unstimulated P29320 -WT lattices did not differ . CONCLUSION : Exposure of intestinal epithelia to the EDGP analog PA stimulates P41180 -dependent P29323 phosphorylation and epithelial-mediated collagen lattice contraction . We speculate that EDGP release within the epithelial layers activates the P41180 receptor , leading to matrix contraction and tissue fibrosis . Dissection of the phenotypic and genotypic associations with nicotinic dependence . INTRODUCTION : Strong evidence demonstrates that nicotine dependence is associated with 4 genetic variants rs16969968 , rs6474412 , rs3733829 , and rs1329650 in large-scale Genome-Wide Association Studies . We examined how these identified genetic variants relate to nicotine dependence defined by different categorical and dimensional measures . METHODS : Four genetic variants were analyzed in 2,047 subjects of European descent ( 1,062 cases and 985 controls ) . DB00184 dependence was assessed with multiple smoking measures , including the Fagerström Test for DB00184 Dependence , the Diagnostic and Statistical Manual for Mental Disorders-IV ( DSM-IV ) nicotine dependence , the DB00184 Dependence Syndrome Scale , and the Wisconsin Inventory of Smoking Dependence Motives . Single-item measures of cigarettes per day ( O75976 ) and time to first cigarette ( Q15669 ) in the morning were also examined . RESULTS : Among the variants , association effect sizes were largest for rs16969968 , with measures of craving and heavy smoking , especially cigarettes smoked per day , showing the largest effects . Significant but weaker associations were found for rs6474412 and rs3733729 but not for rs1329650 . None of the more comprehensive measures of smoking behaviors yielded stronger genetic associations with these variants than did O75976 . CONCLUSIONS : O75976 is an important simple measure that captures in part the genetic associations of P30532 and nicotine dependence , even when other more comprehensive measures of smoking behaviors are examined . The P30532 gene is associated with heavy compulsive smoking and craving ; this should inform the mission to improve the diagnostic validity of DSM-V . UTP induces osteopontin expression through a coordinate action of NFkappaB , activator protein-1 , and upstream stimulatory factor in arterial smooth muscle cells . P10451 ( P10451 ) is an important chemokinetic agent for several cell types . Our earlier studies have shown that its expression is essential for uridine triphosphate ( UTP ) -mediated migration of vascular smooth muscle cells . We demonstrated previously that the activation of an AP-1 binding site located 76 bp upstream of the transcription start in the rat P10451 promoter is involved in the induction of P10451 expression . In this work , using a luciferase promoter deletion assay , we identified a new region of the rat P10451 promoter ( -1837 to -1757 ) that is responsive to UTP . This region contains an NFkappaB site located at -1800 and an Ebox located at -1768 . Supershift electrophoretic mobility shift assay and chromatin immunoprecipitation assays identified NFkappaB and P22415 -1/ P22415 -2 as the DNA binding proteins induced by UTP , respectively , for these two sites . Using dominant negative mutants of O15111 and P22415 transcription factors , we confirmed that NFkappaB and P22415 -1/ P22415 -2 are involved in the UTP-mediated expression of P10451 . Using a pharmacological approach , we demonstrated that P22415 proteins are regulated by the extracellular signal-regulated kinase ( P29323 )1/2 pathway , just as the earlier discovered AP-1 complex , whereas NFkappaB is up-regulated through PKCdelta signals . Finally , our work suggests that the UTP-stimulated P10451 expression involves a coordinate regulation of PKCdelta-NFkappaB , P27361 /2- P22415 , and P27361 /2/NAD(P)H oxidase AP-1 signaling pathways . DB01373 -sensing receptor activation in chronic kidney disease : effects beyond parathyroid hormone control . Secondary hyperparathyroidism ( SHPT ) is an important complication of advanced chronic kidney disease ( CKD ) . DB01012 , an allosteric modulator of the calcium-sensing receptor ( P41180 ) expressed in parathyroid glands , is the only calcimimetic approved to treat SHPT in patients on dialysis . By enhancing P41180 sensitivity for plasma extracellular calcium ( Ca(2+)0 ) , cinacalcet reduces serum parathyroid hormone , Ca(2+)0 , and serum inorganic phosphorous concentrations , allowing better control of SHPT and CKD-mineral and bone disorders . Of interest , the P41180 also is expressed in a variety of tissues where its activation regulates diverse cellular processes , including secretion , apoptosis , and proliferation . Thus , the existence of potential off-target effects of cinacalcet can not be neglected . This review summarizes our current knowledge concerning the potential role(s) of the P41180 expressed in various tissues in CKD-related disorders , independently of parathyroid hormone control . DB00203 inhibits calcineurin/ Q13469 -mediated cyclin A expression in pulmonary artery smooth muscle cells . AIMS : To examine whether calcineurin/NFAT signaling pathway leads to proliferation of pulmonary artery smooth muscle cells ( PASMCs ) by regulating cell cycle proteins and whether the phosphodiesterase-5 ( O76074 ) inhibitor sildenafil affects calcineurin/NFAT-induced cell proliferation . MAIN METHODS : A [(3)H]thymidine incorporation assay was used to examine DNA synthesis ( cell proliferation ) ; cyclin A and Q13469 expressions were determined by Western blot . P24941 ( P24941 ) activity was measured with an in vitro kinase activity assay , and calcineurin and NFAT activity were evaluated using a calcineurin assay kit and a luciferase activity assay , respectively . A chemical inhibitor or siRNA transfection was used to inhibit calcineurin/NFAT signaling pathway . KEY FINDINGS : Serotonin dose-dependently stimulated cyclin A expression in PASMCs . This effect was accompanied by dose-dependent increases in P24941 activity and the rate of DNA synthesis . At the same time , PASMCs treated with serotonin showed dose-dependent activation of calcineurin/NFAT signaling pathway . Inhibition of calcineurin activity by cyclosporine A or loss of Q13469 protein by siRNA transfection abolished serotonin-induced cyclin A expression and consequent P24941 activation and DNA synthesis . We further found that pretreatment of cells with sildenafil suppressed serotonin-triggered activation of calcineurin/ Q13469 signaling pathway and resultant cyclin A expression , P24941 activation and cell proliferation , while the presence of DT-3 [ a specific protein kinase G ( PKG ) peptide inhibitor ] reversed the effects of sildenafil on PASMCs . SIGNIFICANCE : Our study suggests that enhanced PKG activity suppresses calcineurin/ Q13469 cascade-mediated cyclin A expression , P24941 activation and PASMC proliferation to contribute to the overall effects of sildenafil in the treatment of pulmonary hypertension . Regulation of P27361 /2 phosphorylation by acute and chronic morphine - implications for the role of DB02527 -responsive element binding factor ( CREB ) -dependent and Ets-like protein-1 ( Elk-1 ) -dependent transcription ; small interfering RNA-based strategy . Extracellular signal-regulated kinases ( ERKs ) have been shown to be activated by opioids and functionally linked to addiction . Morphine-associated changes in P29323 activity seem to be the characteristic features of opioid action . In this study , we observed a rapid and severe increase in P27361 /2 activity after a 5 min morphine treatment of P29320 -MOR cells ( transfected with the rat mu-opioid receptor P35372 ) expressing mu-opioid receptor . Cellular adaptations to chronic ( 72 h ) morphine treatment were manifested by a slight and sustained increase in P27361 /2 activity . Withdrawal caused by an opioid receptor antagonist - naloxone - attenuated phosphorylation of P27361 /2 . Little information is available on the precise mechanism of P29323 activity regulation . Using RNA interference technology , we generated stably transfected cells with silenced expression of DB02527 -responsive element binding factor ( CREB ) and Ets-like protein-1 ( Elk-1 ) transcription factors , which are known targets for activated P27361 /2 . In these cells , P27361 /2 activity regulation was altered . Silencing of CREB or Elk-1 significantly increased P29323 activation observed after 5 min of morphine stimulation . The initial level of activated ERKs in these cells was also augmented . Moreover , the cellular response to withdrawal signals and chronic opioid treatment was diminished . These differences suggest that both CREB-dependent and Elk-1-dependent transcription contribute to the expression of proteins regulating morphine-induced P29323 activity ( particular phosphatases , upstream kinases or their activatory proteins ) . [ DB01012 -- a new drug for the treatment of secondary hyperparathyroidism in patients with uraemia , parathyroid cancer or primary hyperparathyroidism ] . DB01012 is a new drug with antiparathyroid effects that belongs to the class of calcimimetics . It increases the sensitivity of the calcium-sensing receptor ( P41180 ) to calcium , thus inducing a decrease in plasma parathyroid ( PTH ) levels . In patients with uncontrolled secondary hyperparathyroidism due to uremia , cinacalcet has been shown to decrease the levels of PTH even in those optimally treated with calcium and 1-ahydroxylated vitamin D . DB01012 decreases plasma calcium and plasma PTH levels in patients with primary hyperparathyroidism or parathyroid cancer . New insights into the role of calcium-sensing receptor activation . The discovery of the calcium-sensing receptor ( P41180 ) prompted the identification of substances that affect its function . DB01012 , for example , is a drug that allosterically modifies the receptor so as to increase its sensitivity to circulating calcium ( thus the name " calcimimetic " ) and in this way decreases parathyroid hormone secretion . Clinical use of cinacalcet is already approved for the treatment of primary and secondary hyperparathyroidism , but research is ongoing to identify further potential actions of this drug . The effects and functions of the P41180 have been evaluated in different systems and tissues , beyond parathyroid glands , such arterial walls . A complete understanding of the properties of calcimimetics are of obvious clinical interest , since therapeutic indications may be affected accordingly . P62158 regulates Ca2+-sensing receptor-mediated Ca2+ signaling and its cell surface expression . The Ca(2+)-sensing receptor ( P41180 ) is a member of family C of the GPCRs responsible for sensing extracellular Ca(2+) ( [Ca(2+)](o) ) levels , maintaining extracellular Ca(2+) homeostasis , and transducing Ca(2+) signaling from the extracellular milieu to the intracellular environment . In the present study , we have demonstrated a Ca(2+)-dependent , stoichiometric interaction between P62158 and a P62158 -binding domain ( CaMBD ) located within the C terminus of P41180 ( residues 871-898 ) . Our studies suggest a wrapping around 1-14-like mode of interaction that involves global conformational changes in both lobes of P62158 with concomitant formation of a helical structure in the CaMBD . More importantly , the Ca(2+)-dependent association between P62158 and the C terminus of P41180 is critical for maintaining proper responsiveness of intracellular Ca(2+) responses to changes in extracellular Ca(2+) and regulating cell surface expression of the receptor . S-sulfhydration of Q02750 leads to P09874 activation and DNA damage repair . The repair of DNA damage is fundamental to normal cell development and replication . Hydrogen sulfide ( H2S ) is a novel gasotransmitter that has been reported to protect cellular aging . Here , we show that H2S attenuates DNA damage in human endothelial cells and fibroblasts by S-sulfhydrating Q02750 at cysteine 341 , which leads to P09874 activation . H2S-induced Q02750 S-sulfhydration facilitates the translocation of phosphorylated P27361 /2 into nucleus , where it activates P09874 through direct interaction . Mutation of Q02750 cysteine 341 inhibits P29323 phosphorylation and P09874 activation . In the presence of H2S , activated P09874 recruits P18887 and P49916 to DNA breaks to mediate DNA damage repair , and cells are protected from senescence . The calcium-sensing receptor -- a driver of colon cell differentiation . Dietary Ca(2+) reduces colon cell proliferation and carcinogenesis , but it becomes ineffective or even tumor-promoting during carcinogenesis . It appears that Ca(2+) and the colon cell P41180 together brake the massive cell production in normal colon crypts . The rapid proliferation of the transit-amplifying ( TA ) progeny of the colon stem cells at the bases of the crypts is driven by the " Wnt " signaling mechanism that stimulates proliferogenic genes and prevents apoptogenesis . It appears that TA cell cycling stops and terminal differentiation starts when the cells reach a higher level in the crypt where there is enough external Ca(2+) to stimulate the expression of CaSRs , the signals from which stimulate the expression of P12830 . At this point the P25054 ( adenomatous polyposis coli ) protein appears and some of it enters the nucleus . There it removes the apoptogenesis shield and stops the beta-cateninTcf-4 complex from driving further TA cell proliferation by releasing beta-catenin from the nucleus , and delivering it to cytoplasmic APCaxinGSK-3beta complexes for ultimate proteasomal destruction . Cytoplasmic beta-catenin is prevented from returning to the nucleus by destruction in APCaxinGSK-3beta complexes or locked by the emerging P12830 into adherens junctions which link the cell to proliferatively shut-down functioning cells with P25054 -dependent cytoskeletons moving up and out of the crypt . A common first step in colon carcinogenesis is the loss of functional P25054 which results in the retention of proliferogenic nuclear beta-cateninTcf-4 . This drives the eventual appearance of mutation accumulating , apoptosis-resistant clones the proliferation of which can not be inhibited by external Ca(2+) because of P41180 -disabling gene mutations . Poly( DB02059 )polymerase-1 signalling of the DNA damage induced by P11387 poison in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Glioblastomas are widely characterised by the mutation of the p53 gene and p53 disruption sensitizes glioblastoma cells to P11387 ( TOPO I ) inhibitor-mediated apoptosis . We investigated the effects of combined treatments with the P11387 inhibitor DB01030 and the poly( DB02059 )polymerase-1 inhibitor DB02690 in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Analysis of cell growth and cell cycle kinetics showed a synergistic anti-proliferative effect of 10 nM TPT and 10 microM DB02690 and a G2/M block of the cell cycle . We also evaluated , the influence of TPT+/- DB02690 treatment on P09874 and p53 activity . We got evidences of a TPT-dependent increase of P09874 auto-modification level in both the cells . Moreover , in the D54(p53wt) cells we found that in co-treatments DB02690 incremented the TPT-dependent stimulation of p53 transcriptional activity and increased the P38936 nuclear amount . Conversely , in U251(p53mut) cells we found that DB02690 incremented the TPT-dependent apoptosis characterised by P09874 proteolysis . Our findings suggest that the modulation of P09874 can be considered a strategy in the potentiation of the chemotherapeutic action of TOPO I poisons in glioblastoma cells apart from their p53 status . Modulation of cyclin D1 expression in human tumoral parathyroid cells : effects of growth factors and calcium sensing receptor activation . The study investigated cyclin D1 regulation by growth factors and calcium sensing receptor ( P41180 ) in human tumoral parathyroid cells . Basic fibroblast and epidermal growth factors increased cyclin D1 and phosphorylated extracellular signal-regulated kinases ( pERK1/2 ) levels that were both efficiently inhibited by P41180 agonists . By contrast , in growth factors-free medium cyclin D1 levels were either unaffected or stimulated by P41180 activation independently from P27361 /2 pathway . Transforming growth factor beta ( TGFbeta ) reduced cyclin D1 levels in the majority of tumors , this effect being not influenced by P41180 activation and menin expression levels . In conclusion , in parathyroid tumors cyclin D1 expression was modulated by growth factors and P41180 activation . These data further support the oncogenic role of cyclin D1 , which resulted to be target for stimulation by P09038 and P01133 and inhibition by P41180 and TGFbeta signalling in the parathyroid . Discovery of a calcimimetic with differential effects on parathyroid hormone and calcitonin secretion . Calcimimetics are positive allosteric modulators to the calcium-sensing receptor ( P41180 ) . Activation of the P41180 inhibits the secretion of parathyroid hormone ( PTH ) , stimulates the secretion of calcitonin , and decreases serum calcium ( Ca(2+) ) . DB01012 , a second-generation calcimimetic , is used therapeutically to control PTH in patients with chronic kidney disease who are on dialysis with secondary hyperparathyroidism . A calcimimetic that displays increased separation of PTH versus Ca(2+) lowering in patients would potentially allow the use of calcimimetics to treat patients in earlier stages of renal disease because hypocalcemia can develop in this population . Toward this end , we developed a third-generation calcimimetic , determined the molecular pharmacological properties of it using an operation model of allosteric modulation/agonism , and measured the compound effects on PTH , serum ionized Ca(2+) , and calcitonin levels in 5/6 nephrectomized rats . We found the new molecule effectively reduced PTH levels without promoting calcitonin secretion or hypocalcemia . Furthermore , our third-generation molecule was less efficacious at promoting calcitonin secretion from human thyroid carcinoma cells compared with 3-(2-chlorophenyl)-N- ( ( 1R ) -1-(3-methoxyphenyl)ethyl ) -1-propanamine ( R-568 ) , a first-generation calcimimetic . These data provide evidence that calcimimetics with increased potency can be used to lower PTH without production of significant hypocalcemia because the threshold for inhibition of PTH secretion is much lower than the threshold for calcitonin secretion . Calcimimetics in the chronic kidney disease-mineral and bone disorder . Mineral and bone disorders ( MBD ) are both an early and very common complication of chronic kidney disease ( CKD ) . It is now accepted that they represent a significant risk factor , explaining the high cardiovascular morbidity and mortality in CKD patients . During the last decade , we have been witnessing many advances in the nomenclature , classification , pathophysiology , diagnosis , and treatment of CKD and some of its complications , such as CKD-MBD . The identification of the calcium-sensing receptor ( P41180 ) involvement in the pathogenesis of primary and secondary hyperparathyroidism ( SHPT ) and the availability of a new class of drugs called calcimimetics are two outstanding examples . DB01012 , the only available calcimimetic , has been shown to be a very effective therapeutic tool in CKD-MBD . Many clinical trials with cinacalcet in hemodialysis patients with SHPT have shown a reduction in parathyroid hormone , calcium ( Ca ) , phosphate ( P ) and Ca x P product levels , allowing far greater success in reaching therapeutic goals as recommended by international guidelines . Additionally , some studies have shown that the use of cinacalcet may improve other aspects of CKD-MBD , reducing the risk of vascular calcification and parathyroidectomy , among others . Prospective studies on dialysis patients , with hard endpoint data , are currently underway . This review summarizes the most significant aspects of calcimimimetics based on both experimental and clinical results , underlining their possibilities not only for the treatment of isolated SHPT but also for other CKD-MBD related conditions . DB01373 -sensing receptor gene polymorphism Arg990Gly and its possible effect on response to cinacalcet HCl . DB01012 , a novel calcimimetic compound , is effective in reducing parathyroid hormone ( PTH ) levels in approximately 70 % of patients with secondary hyperparathyroidism . However , interindividual variations in the dose required to achieve the treatment goal have been noted in clinical studies . Our investigation examined the genetic polymorphisms of the calcium-sensing receptor ( P41180 ) gene as one possible cause of the different responses to cinacalcet . We report data on seven end-stage renal failure patients who were treated with regular haemodialysis and who participated in clinical trials of cinacalcet . All patients had secondary hyperparathyroidism with baseline intact PTH ( iPTH ) levels greater than 600 pg/ml . Three patients were male and four female with a mean+/-SD age of 60+/-12 years . DNA was extracted from peripheral lymphocytes . An area in exon 7 of the P41180 gene was amplified by the polymerase chain reaction and sequenced . Mean+/-SD baseline iPTH was 1086+/-189 pg/ml . The five patients without Arg990Gly demonstrated a 29.7+/-4.0 % ( +/-SEM ) reduction in iPTH from individual baseline . One patient was found to be homozygous for the Arg990Gly polymorphism and another was heterozygous for both arginine and glycine alleles . The homozygous patient showed a significantly higher sensitivity to cinacalcet compared to the other patients ( P=0.003 ) with a 76.3+/-7.7 % reduction in iPTH from baseline . No polymorphisms were noted in codons 986 or 1011 . This preliminary study points to the possibility that patients homozygous for glycine at the 990 position in exon 7 of the P41180 may be more sensitive to the calcimimetic drug cinacalcet compared to those who are homozygous for arginine at that location . G protein-coupled receptors disrupted in human genetic disease . Genetic variation in G protein-coupled receptors ( GPCRs ) results in the disruption of GPCR function in a wide variety of human genetic diseases . In vitro strategies have been used to elucidate the molecular pathologies that underlie naturally occurring GPCR mutations . Various degrees of inactive , overactive , or constitutively active receptors have been identified . These mutations often alter ligand binding , G protein coupling , receptor desensitization , and receptor recycling . The role of inactivating and activating calcium-sensing receptor ( P41180 ) mutations is discussed with respect to familial hypocalciuric hypercalemia ( FHH ) and autosomal dominant hypocalemia ( DB00067 ) . Among DB00067 mutations , those associated with tonic-clonic seizures are discussed . Other receptors discussed include rhodopsin , thyrotropin , parathyroid hormone , melanocortin , follicle-stimulating hormone , luteinizing hormone , gonadotropin-releasing hormone ( GnRHR ) , adrenocorticotropic hormone , vasopressin , endothelin-beta , purinergic , and the G protein associated with asthma ( Q6W5P4 ) . Diseases caused by mutations that disrupt GPCR function are significant because they might be selectively targeted by drugs that rescue altered receptors . Examples of drug development based on targeting GPCRs mutated in disease include the calcimimetics used to compensate for some P41180 mutations , obesity therapeutics targeting melanocortin receptors , interventions that alter GnRHR loss from the cell surface in idiopathic hypogonadotropic hypogonadism and novel drugs that might rescue the Q9H244 receptor in a rare bleeding disorder . The discovery of Q6W5P4 suggests that drug screens against variant GPCRs may identify novel drugs . This review of the variety of GPCRs that are disrupted in monogenic disease provides the basis for examining the significance of common pharmacogenetic variants . DB01012 upregulates calcium-sensing receptors of parathyroid glands in hemodialysis patients . BACKGROUND : DB01012 hydrochloride ( cinacalcet ) , a calcimimetic , has been shown to upregulate calcium-sensing receptor ( P41180 ) expression in parathyroid glands of rats with chronic renal insufficiency . However , the effect of cinacalcet on the reduced P41180 expression in human parathyroid glands remains to be elucidated . METHODS : Four normal parathyroid glands and 71 hyperplastic parathyroid glands from 18 hemodialysis patients with refractory secondary hyperparathyroidism ( SHPT ) treated with ( n = 10 ; cinacalcet group ) or without ( n = 8 ; conventional group ) cinacalcet were examined immunohistochemically with a specific antibody against P41180 . The expression level of P41180 was analyzed semiquantitatively . RESULTS : Compared with normal glands , the immunohistochemical expression of P41180 was decreased significantly in both the cinacalcet and conventional groups . In the cinacalcet group , the expression of P41180 was increased significantly compared with that in the conventional group ( 1.83 ± 0.14 vs. 0.87 ± 0.15 , p < 0.001 ) , even though the proportion of patients using vitamin D sterols and the mean administered dose of calcitriol equivalents were not significantly different between the two groups . The expression of P41180 was significantly decreased in the larger glands ( > 500 mg ) compared with that in the smaller glands ( < 500 mg ) in both groups ; furthermore , it was markedly decreased in areas of nodular hyperplasia compared with diffuse hyperplasia in the cinacalcet group . CONCLUSIONS : Our results indicate that cinacalcet upregulates the depressed expression of P41180 in hemodialysis patients with SHPT , and that insufficient expression of P41180 , especially in larger glands with advanced nodular hyperplasia , underlies the pathogenesis of SHPT in patients who are resistant to cinacalcet . Pharmacogenomics of methadone maintenance treatment . DB00333 is the major opioid substitution therapy for opioid dependence . Dosage is highly variable and is often controlled by the patient and prescriber according to local and national policy and guidelines . Nevertheless many genetic factors have been investigated including those affecting its metabolism ( P20813 -consistent results ) , efflux transport ( P-gp-inconsistent results ) , target μ-opioid receptor ( μ-opioid receptor-inconsistent results ) and a host of other receptors ( P14416 ) and signaling elements ( P48051 and P32121 ; not replicated ) . None by themselves have been able to substantially explain dosage variation ( the major but not sole end point ) . When multiple genes have been combined such as P08183 , P20813 , P35372 and P14416 a greater contribution to dosage variation was found but not as yet replicated . As stabilization of dosage needs to be made rapidly , it is imperative that larger internationally based studies be instigated so that genetic contribution to dosage can be properly assessed , which may or may not tailor to different ethnic groups and each country 's policy towards an outcome that benefits all . Novel strategies in drug discovery of the calcium-sensing receptor based on biased signaling . A hallmark of chronic kidney disease is hyperphosphatemia due to renal phosphate retention . Prolonged parathyroid gland exposure to hyperphosphatemia leads to secondary hyperparathyroidism characterized by hyperplasia of the glands and excessive secretion of parathyroid hormone ( PTH ) , which causes renal osteodystrophy . PTH secretion from the parathyroid glands is controlled by the calcium-sensing receptor ( P41180 ) that senses extracellular calcium . High extracellular calcium activates the P41180 causing inhibition of PTH secretion through multiple signaling pathways . DB01012 is the first drug targeting the P41180 and can be used to effectively control and reduce PTH secretion in PTH-related diseases . DB01012 is a positive allosteric modulator of the P41180 and affects PTH secretion from parathyroid glands by shifting the calcium-PTH concentration-response curve to the left . One major disadvantage of cinacalcet is its hypocalcemic side effect , which may be caused by increased P41180 -mediated calcitonin secretion from the thyroid gland . However , multiple studies indicate that PTH and calcitonin secretion are stimulated by different signaling pathways , and therefore it might be possible to develop a P41180 activating drug that selectively activates signaling pathways that inhibit PTH secretion while having no effect on signaling pathways involved in calcitonin secretion . Such a drug would have the same therapeutic value as cinacalcet in lowering PTH secretion while eliminating the side effect of hypocalcemia by virtue of it not affecting calcitonin secretion . The present review will focus on recent advancements in understanding signaling and biased signaling of the P41180 , and how that may be utilized to discover new and smarter drugs targeting the P41180 . Gq-mediated Akt translocation to the membrane : a novel PIP3-independent mechanism in platelets . Akt is an important signaling molecule regulating platelet aggregation . Akt is phosphorylated after translocation to the membrane through Gi signaling pathways by a phosphatidylinositol-3,4,5-trisphosphate ( PIP3 ) -dependent mechanism . However , Akt is more robustly phosphorylated by thrombin compared with adenosine 5'-diphosphate in platelets . This study investigated the mechanisms of Akt translocation as a possible explanation for this difference . Stimulation of washed human platelets with protease-activated receptor agonists caused translocation of Akt to the membrane rapidly , whereas phosphorylation occurred later . The translocation of Akt was abolished in the presence of a Gq-selective inhibitor or in Gq-deficient murine platelets , indicating that Akt translocation is regulated downstream of Gq pathways . Interestingly , phosphatidylinositol 3-kinase ( PI3K ) inhibitors or Q9H244 antagonist abolished Akt phosphorylation without affecting Akt translocation to the membrane , suggesting that Akt translocation occurs through a PI3K/PIP3/Gi-independent mechanism . An Akt scaffolding protein , P38936 -activated kinase ( PAK ) , translocates to the membrane after stimulation with protease-activated receptor agonists in a Gq-dependent manner , with the kinetics of translocation similar to that of Akt . Coimmunoprecipitation studies showed constitutive association of PAK and Akt , suggesting a possible role of PAK in Akt translocation . These results show , for the first time , an important role of the Gq pathway in mediating Akt translocation to the membrane in a novel Gi/PI3K/PIP3-independent mechanism . [ Medical treatment of primary hyperparathyroidism : role of calcimimetics ] . Primary hyperparathyroidism ( PHPT ) is a common endocrinological process , characterized by chronic elevation of serum concentrations of calcium and parathyroid hormone ( PTH ) . The only intervention able to cure the disease is parathyroidectomy . However , there are few valid medical alternatives for patients whose PHPT is unresolved by surgery , or in those with contraindications for surgery or who refuse the procedure . The discovery of calcium-sensing receptors ( CaSRs ) , which regulate PTH secretion according to extracellular calcium concentrations , has allowed specific anti-parathyroid drugs called calcimimetics to be designed . DB01012 is an allosteric modulator of P41180 that has demonstrated safety and efficacy in controlling serum calcium values and in reducing PTH levels in patients with PHPT . The exact role of calcimimetics in the overall management of PHPT is promising and should be considered in future clinical practice guidelines . Genetics of physical activity and physical inactivity in humans . Emerging evidence suggests that physical activity and sedentary behavior [ reflected in physical inactivity ( PI ) ] , might be two different phenotypes that may have distinct underlying physiological mechanisms . The purpose of this review is to summarize the existing literature on the genetic determinants of PA and PI phenotypes in humans , considering them as distinct behaviors . Completed in January 2011 , this review includes family studies , twin studies , association studies , genome-wide linkage studies and genome-wide association scan ( GWAs ) reporting different physical activity/inactivity-related phenotypes . In regards to PA , familial aggregation studies resulted in heritability estimates ranging from 0 to 60 % , and twin studies yielded heritability estimates ( a(2) ) and shared environment ( c(2) ) scores for PA phenotypes ranging from 0.00 to 0.85 and 0.00 to 0.84 , respectively . Unique environmental ( e(2) ) results are well dispersed from 0.12 to 0.72 . Suggestive linkages were found with markers nearby different activity-related genes : P24530 , P32245 , P25874 , P12104 , P41180 , Q8IVB4 . Significant associations with PA phenotypes were found for Ace , Gln223ARrg , P32245 and P14416 genes . We found one GWAs that reported novel SNPs in the O95340 gene on chromosome 10q23.2 and in two intergenic regions on chromosomes 2q33.1 and 18p11.32 . Heritability estimates for PI ranged from 25 to 60 % and linkage studies recorded higher LOD scores for PI versus PA . The P12821 genotype was strongly associated with PI . There are potentially different genetic influences on PA versus PI phenotypes . Future studies should focus on the different genetic influences on PA and PI to improve our understanding of underlying determinants of these behaviors . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . [ Vascular Calcification - Pathological Mechanism and Clinical Application - . The effect of cinacalcet on vascular calcification ] . DB01012 acts on calcium receptors ( CaR ) expressed on chief cells of the parathyroid gland to inhibit the secretion of parathyroid hormone ( PTH ) . This drug inhibits PTH secretion without causing an elevation of serum calcium and phosphorus , unlike active vitamin D . Several experimental studies demonstrated an inhibitory effect of calcimimetics on the progression of vascular calcification in animals with chronic kidney disease ( CKD ) , in keeping with the expression of the calcium-sensing receptor ( P41180 ) in vascular tissue . The EVOLVE , evaluated in patients with CKD 5D the effects of the cinacalcet on the progression of vascular calcification and hard cardiovascular outcomes , respectively . The EVOLVE trials missed their respective primary end point by intent-to-treat analysis . However , recently , in order to define the frequency of fatal and nonfatal cardiovascular events attributable to atherosclerotic and nonatherosclerotic mechanisms , risk factors for these events , and the effects of cinacalcet , post hoc analysis using adjudicated data collected during the EVOLVE Trial were perfomed . In this trial , combining fatal and nonfatal cardiovascular events , randomization to cinacalcet reduced the rates of sudden death and heart failure . Patients randomized to cinacalcet experienced fewer nonatherosclerotic cardiovascular events , while the effect of cinacalcet on atherosclerotic events did not reach statistical significance . DB01012 Treatment in an Adolescent With Concurrent 22q11.2 Deletion Syndrome and Familial Hypocalciuric Hypercalcemia Type 3 Caused by P53680 Mutation . CONTEXT : The 22q11.2 deletion syndrome ( DS ) is a common multiple anomaly syndrome in which typical features include congenital heart defects , facial dysmorphism , and palatal anomalies . Hypocalcemia due to hypoparathyroidism is a common endocrine manifestation resulting from variable parathyroid hypoplasia , but hypercalcemia has not previously been reported in 22q11.2 DS . CASE DESCRIPTION : Our patient is a 16-year-old adolescent male with dysmorphic facial features and delayed motor and speech development . At 2 years of age , 22q11.2 DS was confirmed by fluorescence in situ hybridization . In contrast to hypoparathyroidism that is usually seen in 22q11.2 DS , this patient had early childhood-onset hypercalcemia with inappropriately high PTH levels and hypocalciuria . Genomic DNA was obtained from the proband and screened for calcium-sensing receptor ( P41180 ) mutations with negative results . No parathyroid tissue could be localized by imaging or surgical exploration . As a result of symptomatic hypercalcemia , the patient was treated with a calcimimetic ( cinacalcet ) . During the treatment , plasma calcium normalized with mild symptoms of hypocalcemia . After discontinuation of cinacalcet , calcium returned to high pretreatment levels . Further DNA analysis of adaptor protein-2 σ subunit ( P53680 ) showed a heterozygous missense mutation c.44 G > T , resulting in a p.R15L substitution ; the mutation was absent in the healthy parents and two siblings . CONCLUSIONS : Hypercalcemia in our patient with 22q11.2 DS could be explained by the de novo mutation in P53680 . Identification of a genetic cause for hypercalcemia is helpful in guiding management and avoiding unnecessary treatment . Behind the curtain : cellular mechanisms for allosteric modulation of calcium-sensing receptors . DB01373 -sensing receptors ( P41180 ) are integral to regulation of systemic Ca(2+) homeostasis . Altered expression levels or mutations in P41180 cause Ca(2+) handling diseases . P41180 is regulated by both endogenous allosteric modulators and allosteric drugs , including the first Food and Drug Administration-approved allosteric agonist , DB01012 HCl ( Sensipar® ) . Recent studies suggest that allosteric modulators not only alter function of plasma membrane-localized P41180 , but regulate P41180 stability at the endoplasmic reticulum . This brief review summarizes our current understanding of the role of membrane-permeant allosteric agonists in cotranslational stabilization of P41180 , and highlights additional , indirect , signalling-dependent role(s) for membrane-impermeant allosteric drugs . Overall , these studies suggest that allosteric drugs act at multiple cellular organelles to control receptor abundance and hence function , and that drug hydrophobicity can bias the relative contributions of plasma membrane and intracellular organelles to P41180 abundance and signalling . Q16739 -100 , a novel galectin-3 antagonist , modulates Q8WXI8 -1 , Q13794 , and cell cycle to induce myeloma cell death . Q16739 -100 is a galectin-3 antagonist with an acceptable human safety profile that has been demonstrated to have an antimyeloma effect in the context of bortezomib resistance . In the present study , the mechanisms of action of Q16739 -100 are elucidated in myeloma cell lines and primary tumor cells . Q16739 -100 induced inhibition of proliferation , accumulation of cells in sub-G(1) and G(1) phases , and apoptosis with activation of both caspase-8 and -9 pathways . Dose- and time-dependent decreases in Q8WXI8 -1 and BCL-X(L) levels also occurred , accompanied by a rapid induction of Q13794 protein , whereas BCL-2 , Q07812 , Q16611 , O43521 , Q92934 , P55957 , and PUMA remained unchanged . The cell-cycle inhibitor P38936 (Cip1) was up-regulated by Q16739 -100 , whereas the procycling proteins CYCLIN E2 , CYCLIN D2 , and Q00534 were all reduced . Reduction in signal transduction was associated with lower levels of activated P25963 , O15111 , and AKT as well as lack of P25963 and AKT activation after appropriate cytokine stimulation ( insulin-like growth factor-1 , tumor necrosis factor-alpha ) . Primary myeloma cells showed a direct reduction in proliferation and viability . These data demonstrate that the novel therapeutic molecule , Q16739 -100 , is a potent modifier of myeloma cell biology targeting apoptosis , cell cycle , and intracellular signaling and has potential for myeloma therapy . PPARgamma activation abolishes LDL-induced proliferation of human aortic smooth muscle cells via SOD-mediated down-regulation of superoxide . Native LDL would be a mitogenic and chemotactic stimulus of VSMC proliferation and differentiation in the atherosclerotic lesion where endothelial disruption occurred . In previous studies , our group investigated the molecular mechanisms by which LDL induces P10145 production and by which PPARalpha activation abolishes LDL effects in human aortic SMCs ( hAoSMCs ) . Herein is the first report of PPARgamma activation by troglitazone ( TG ) exerting its inhibitory effects on LDL-induced cell proliferation via generation not of H(2)O(2) , but of O2(.-) , and the subsequent activation of Erk1/2 in hAoSMCs . Moreover , in this study TG abolished the LDL-accelerated G(1)-S progression to control levels via down-regulation of active cyclinD1/ P11802 and cyclinE/ P24941 complexes and up-regulation of P38936 (Cip1) expression . TG exerted its anti-proliferative effects through the up-regulation of basal superoxide dismutase ( SOD ) expression . This data suggests that the regulation of O2(.-) is located at the crossroads between LDL signaling and cell proliferation . P38936 -activated kinase-aberrant activation and translocation in Alzheimer disease pathogenesis . Defects in dendritic spines and synapses contribute to cognitive deficits in mental retardation syndromes and , potentially , Alzheimer disease . P38936 -activated kinases ( PAKs ) regulate actin filaments and morphogenesis of dendritic spines regulated by the Rho family GTPases Rac and Cdc42 . We previously reported that active PAK was markedly reduced in Alzheimer disease cytosol , accompanied by downstream loss of the spine actin-regulatory protein Q16643 . beta-Amyloid ( Abeta ) oligomer was implicated in PAK defects . Here we demonstrate that PAK is aberrantly activated and translocated from cytosol to membrane in Alzheimer disease brain and in 22-month-old Tg2576 transgenic mice with Alzheimer disease . This active PAK coimmunoprecipitated with the small GTPase Rac and both translocated to granules . Abeta42 oligomer treatment of cultured hippocampal neurons induced similar effects , accompanied by reduction of dendrites that were protected by kinase-active but not kinase-dead PAK . Abeta42 oligomer treatment also significantly reduced N-methyl-d-aspartic acid receptor subunit Q13224 phosphotyrosine labeling . The Src family tyrosine kinase inhibitor Q99463 significantly blocked the PAK/Rac translocation but not the loss of p- Q13224 in Abeta42 oligomer-treated neurons . Src family kinases are known to phosphorylate the Rac activator Tiam1 , which has recently been shown to be Abeta-responsive . In addition , anti-oligomer curcumin comparatively suppressed PAK translocation in aged Tg2576 transgenic mice with Alzheimer amyloid pathology and in Abeta42 oligomer-treated cultured hippocampal neurons . Our results implicate aberrant PAK in Abeta oligomer-induced signaling and synaptic deficits in Alzheimer disease . Xaliproden ( SR57746A ) induces P08908 receptor-mediated Q96HU1 kinase activation in PC12 cells . Neurotrophic growth factors are involved in cell survival . However , natural growth factors have a very limited therapeutic use because of their short half-life . In the present study , we investigated the mechanism of action of a non-peptidic neurotrophic drug , Xaliproden , a potential molecule for the treatment of motoneuron diseases , since the transduction pathways of this synthetic P08908 agonist are very poorly understood . Xaliproden does not activate the Trk receptor but causes a rapid increase in the activities of the P27361 and P28482 isoforms of Q96HU1 kinase , which then rapidly decrease to the basal level . We demonstrate that isoforms of the P29353 adapter protein are phosphorylated independently of each other and are probably not the source of the Xaliproden-induced Q96HU1 kinases activation . The inhibitor of Ras farnesylation , FPT-1 , and the protein kinase C inhibitors , GF 109203X and chelerythrine , inhibited the Xaliproden-induced Q96HU1 kinase activation , suggesting p21Ras and PKC involvement . Moreover , the observations that the P08908 antagonist , pindobind , and pertussis toxin abolished the Xaliproden-induced P29323 stimulation suggested that Xaliproden activates the Q96HU1 kinase pathways by stimulating the G protein-coupled receptor , P08908 . These results demonstrate clearly that the non-peptidic compound , Xaliproden , exerts its neurotrophic effects through a mechanism of action differing from that of neurotrophins . These findings suggest that this compound does not involve MAPK activation by TrkA receptor stimulation but acts by Q96HU1 kinase pathway by a pertussis toxin-sensitive mechanism involving P08908 receptors , P38936 Ras and MEK-1 and by PKC and Akt pathways . Overexpression of SnoN/SkiL , amplified at the 3q26.2 locus , in ovarian cancers : a role in ovarian pathogenesis . High-resolution array comparative genomic hybridization of 235 serous epithelial ovarian cancers demonstrated a regional increase at 3q26.2 encompassing SnoN/SkiL , a coregulator of SMAD/TGFbeta signaling . SnoN RNA transcripts were elevated in approximately 80 % of advanced stage serous epithelial ovarian cancers . In both immortalized normal ( TIOSE ) and ovarian carcinoma cell lines ( OVCA ) , SnoN RNA levels were increased by TGFbeta stimulation and altered by LY294002 and JNK II inhibitor treatment suggesting that the PI3K and JNK signaling pathways may regulate TGFbeta-induced increases in SnoN RNA . In TIOSE , SnoN protein levels were reduced 15min post TGFbeta-stimulation , likely by proteosome-mediated degradation . In contrast , in OVCA , SnoN levels were elevated 3h post-stimulation potentially as a result of inhibition of the proteosome . To elucidate the role of SnoN in ovarian tumorigenesis , we explored the effects of both increasing and decreasing SnoN levels . In both TIOSE and OVCA , SnoN siRNA decreased cell growth between 20 and 50 % concurrent with increased P38936 levels . In TIOSE , transient expression of SnoN repressed TGFbeta induction of P05121 promoters with little effect on the P38936 promoter or resultant cell growth . In contrast to the effects of transient expression , stable expression of SnoN in TIOSE led to growth arrest through induction of senescence . Collectively , these results implicate SnoN levels in multiple roles during ovarian carcinogenesis : promoting cellular proliferation in ovarian cancer cells and as a positive mediator of cell cycle arrest and senescence in non-transformed ovarian epithelial cells . [ Prominent features of management strategies in acute coronary syndromes with the new oral antiplatelet agents ] . The novel oral Q9H244 inhibitors ( prasugrel and ticagrelor ) have been incorporated into the recently updated acute coronary syndrome ( ACS ) guidelines , as an adjunct antiplatelet treatment to aspirin . The studies involving the use of new oral antiplatelet agents that are more potent , predictable and faster platelet inhibitors than clopidogrel have demonstrated superiority with respect to the primary composite endpoint ( cardiovascular death , non-lethal myocardial infarction , stroke ) for both prasugrel and ticagrelor compared to clopidogrel . The subgroup analysis of the relevant studies showed that these new agents differ in their level of efficacy in different ACS patient subgroups : ( 1 ) Mortality was reduced with ticagrelor ; ( 2 ) DB08816 is especially more effective in intermediate-and high-risk non-ST elevation ACS patients in whom early invasive strategy is selected ; ( 3 ) Prasugrel should be especially preferred in patients with acute ST elevation myocardial infarction undergoing percutaneous coronary intervention ( P05154 ) after diagnostic angiography ; and ( 4 ) Prasugrel is more effective in diabetic patients . While clopidogrel is recommended for ACS patients who are followed with a non-invasive strategy or who have not undergone percutaneous revascularization , it is the last line choice or an alternative to the Q9H244 inhibitor therapy for patients undergoing invasive strategy . Agonists of the Q9H244 -receptor activate Q96HU1 kinase by a ras-independent pathway in rat P13671 glioma . We have previously shown that an ecto-NPPase modulates the DB00171 - and ADP-mediated Q9H244 -receptor activation in rat P13671 glioma . In the present study , 2MeSADP and Ap(3)A induced no detectable PI turnover and were identified as specific agonists of the Q9H244 -receptor with EC(50) values of 250 +/- 37 pM and 1 +/- 0.5 microM , respectively . Q9H244 -receptor stimulation increased Q96HU1 kinase ( P27361 /2 ) activation that returned to the basal level 4 h after stimulation and was correlated with a gradual desensitization of the Q9H244 -purinoceptor . The purinoceptor antagonists DIDS and Q08999 blocked Q96HU1 kinase activation . An IP(3)-independent Ca(2+)-influx was observed after Q9H244 -receptor activation . Inhibition of this influx by Ca(2+)-chelation , did not affect Q96HU1 kinase activation . Pertussis toxin , toxin B , selective PKC-inhibitors and a specific MEK-inhibitor inhibited the 2MeSADP- and Ap(3)A-induced Q96HU1 kinase activation . In addition , transfection with dominant negative RhoA(Asn19) rendered P13671 cells insensitive to Q9H244 -receptor-mediated Q96HU1 kinase activation whereas dominant negative ras was without effect . Immunoprecipitation experiments indicated a significant increase in the phosphorylation of raf-1 after Q9H244 -receptor activation . We may conclude that Q9H244 -purinoceptor agonists activate Q96HU1 kinase through a G(i)-RhoA-PKC-raf-MEK-dependent , but ras- and Ca(2+)-independent cascade . Pharmacology of the calcium sensing receptor . DB01373 sensing receptor ( P41180 ) is a G-protein couple receptor which plays a key role in calcium homeostasis in vertebrates . Its extracellular domain is sensitive to divalent cations , aminoacids and polyamines . In parathyroid glands , P41180 activation causes parathyroid hormone ( PTH ) reduction and subsequently a decrease in blood calcium concentration . In PTH-dependent disorders , e.g. primary and secondary hyperparathyroidism ( Q9HD23 ) , the need for therapeutic options other than surgery led to the synthesis of various allosteric P41180 agonists ( calcimimetics ) , such as cinacalcet . DB01012 is the only calcimimetic approved for Q9HD23 secondary to chronic kidney disease ( CDK ) , parathyroid carcinoma , and , in some countries , primary Q9HD23 . Clinical trials showed that cinacalcet reduced PTH and calcemia both in CDK and primary Q9HD23 , lowering the risk of bone fractures , surgery , and cardiovascular complications in the former patients . Long-term safety and pharmacoeconomics have to be fully tested yet . Few both in vitro and in vivo studies showed an association between Arg990Gly- P41180 polymorphism and cinacalcet sensitivity , though in patients with severe P41180 inactivating mutations the drug substantially retained its positive clinical effects . Recently , a new class of allosteric antagonists of P41180 , i.e. calcilytics , has been synthesized . Calcilytics are structurally similar to calcimimetics , but exert their effects acting on a different allosteric site . Infusion of calcilytics was followed by transient rise in PTH and calcium . One of these compounds , DB05255 , was able to increase femur BMD in post menopausal women , but with induction of mild hyperparathyroidism . In the future , calcilytics may contribute to the osteoporosis treatment choice . Successful use of bisphosphonate and calcimimetic in neonatal severe primary hyperparathyroidism . Neonatal primary hyperparathyroidism ( NPHT ) is associated with an inactivating homozygous mutation of the calcium sensing receptor ( P41180 ) . The P41180 is expressed most abundantly in the parathyroid glands and the kidney and regulates calcium homeostasis through its ability to modulate parathormone secretion and renal calcium reabsorption . NPHT leads to life threatening hypercalcemia , nephrocalcinosis , bone demineralization , and neurologic disabilities . Surgery is the treatment of choice . While waiting for surgery , bisphosphonates offer a good alternative to deal with hypercalcemia . DB01012 is a class II calcimimetic that increases P41180 affinity for calcium , leading to parathormone suppression and increased calcium renal excretion . At present , there is little evidence as to whether cinacalcet could improve the function of mutant P41180 in NPHT . We report a case of NPHT , treated successfully with bisphosphonates and cinacalcet after surgery failure . To our knowledge , it is the first time cinacalcet has been used for NPHT . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . P35372 and P20813 gene variants as risk factors in methadone-related deaths . DB00333 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 and micro-opioid receptor ( P35372 ) 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 *4,*9 , and *6 alleles and the P35372 A118G variant was determined by SNP genotyping . P20813 *4 , *9 , and *6 alleles were found to be associated with higher postmortem methadone concentrations in blood ( P < or = 0.05 ) . P35372 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 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 *6 and A118G . Clinical utility of calcimimetics targeting the extracellular calcium-sensing receptor ( P41180 ) . Calcimimetics , which activate the extracellular calcium ( Ca(o)(2+) ) -sensing receptor in the parathyroid and other tissues participating in Ca(o)(2+) homeostasis , were the first described allosteric activators of a G-protein-coupled receptor . DB01012 , the only calcimimetic currently approved for human use , is used clinically for treating secondary hyperparathyroidism ( e.g. , overactivity of parathyroid glands ) in patients being dialyzed for chronic kidney disease . By sensitizing the parathyroids to Ca(o)(2+) , cinacalcet lowers the circulating parathyroid hormone ( PTH ) level . It also reduces serum calcium and phosphate , changes increasing the percentage of patients achieving the guidelines recommended by the National Kidney Foundation ( NKF ) for these minerals . Studies are underway addressing whether better adherence to these guidelines in patients receiving cinacalcet reduces cardiovascular disease and related mortality , which are both common is the dialysis population . The second approved use of cinacalcet is for treating hypercalcemia in patients with inoperable parathyroid carcinoma . In this setting , it provides the first medical therapy chronically lowering serum calcium concentration in this condition , albeit not to normal in most patients . Its effect on the long-term prognosis of these patients , if any , is presently unclear . " Off-label " administration of cinacalcet [ i.e. , not yet approved by the US Food and Drug Administration ( FDA ) ] effectively lowers serum calcium and/or PTH in various other forms of hyperparathyroidism and increases serum phosphate in renal phosphate-wasting syndromes by reducing PTH-induced phosphaturia . In the future , the drug could conceivably be utilized to modulate the activity of the P41180 in other tissues ( i.e. , kidney , colon ) in therapeutically desirable ways . Calcimimetic and calcilytic drugs for treating bone and mineral-related disorders . The calcium-sensing receptor ( P41180 ) plays a pivotal role in regulating systemic Ca(2+) homeostasis and is a target for drugs designed to treat certain disorders of bone and mineral metabolism . Calcimimetics are agonists or positive allosteric modulators of the P41180 ; they inhibit parathyroid hormone ( PTH ) secretion and stimulate renal Ca(2+) excretion . The first calcimimetic drug is cinacalcet , a positive allosteric modulator of the P41180 that is approved for treating secondary hyperparathyroidism ( Q9HD23 ) in patients on renal replacement therapy and for some forms of primary Q9HD23 characterized by clinically significant hypercalcemia . DB01012 is also being investigated as a therapy for other hypercalcemic conditions and certain hypophosphatemic disorders . Calcilytics are P41180 inhibitors that stimulate the secretion of PTH and decrease renal excretion of Ca(2+) . Although calcilytics have failed thus far as anabolic therapies for osteoporosis , they are currently being evaluated as novel therapies for new indications involving hypocalcemia and/or hypercalciuria . Suppression of NF-kappaB activity by sulfasalazine is mediated by direct inhibition of IkappaB kinases alpha and beta . BACKGROUND & AIMS : Activation of NF-kappaB/Rel has been implicated in the pathogenesis of inflammatory bowel disease ( Q9UKU7 ) . Various drugs used in the treatment of Q9UKU7 , such as glucocorticoids , DB00244 , and sulfasalazine , interfere with NF-kappaB/Rel signaling . The aim of this study was to define the molecular mechanism by which sulfasalazine inhibits NF-kappaB activation . METHODS : The effects of sulfasalazine and its moieties on NF-kappaB signaling were evaluated using electromobility shift , transfection , and immune complex kinase assays . The direct effect of sulfasalazine on O15111 ( IKK ) activity was investigated using purified recombinant O15111 and -beta proteins . RESULTS : NF-kappaB/Rel activity induced by tumor necrosis factor alpha , 12-O-tetradecanoylphorbol-13-acetate , or overexpression of NF-kappaB-inducing kinase , O15111 , O14920 , or constitutively active O15111 and O14920 mutants was inhibited dose dependently by sulfasalazine . Sulfasalazine inhibited tumor necrosis factor alpha-induced activation of endogenous IKK in Jurkat T cells and SW620 colon cells , as well as the catalytic activity of purified O15111 and O14920 in vitro . In contrast , the moieties of sulfasalazine , DB00244 , and sulfapyridine or DB00233 had no effect . Activation of extracellular signal-related kinase ( P29323 ) 1 and 2 , c-Jun-N-terminal kinase ( JNK ) 1 , and p38 was unaffected by sulfasalazine . The decrease in substrate phosphorylation by O15111 and -beta is associated with a decrease in autophosphorylation of IKKs and can be antagonized by excess adenosine triphosphate . CONCLUSIONS : These data identify sulfasalazine as a direct inhibitor of O15111 and -beta by antagonizing adenosine triphosphate binding . The suppression of NF-kappaB activation by inhibition of the IKKs contributes to the well-known anti-inflammatory and immunosuppressive effects of sulfasalazine . Clinical utilization of cinacalcet in hypercalcemic conditions . INTRODUCTION : DB01012 has recently been introduced as a treatment for secondary hyperparathyroidism in dialysis patients and for parathyroid carcinoma . However , there has been an increasing interest in finding out whether cinacalcet can be used as a treatment for other parathyroid hormone ( PTH ) -dependent hypercalcemic conditions also . AREAS COVERED : The article reports the most relevant recent contributions dealing with calcium sensing receptor ( P41180 ) physiology as well as cinacalcet pharmacokinetics and pharmacodynamics . It also looks at the different hypercalcemic conditions where the use of cinacalcet has been proposed . This article was researched using clinical trials , case reports and outstanding basic research published in the last 3 years ( MEDLINE database up to 31 November 2010 ) . It provides the reader with an insight into the many unaddressed issues regarding cinacalcet that need to be resolved before it can be used in newly proposed fields . EXPERT OPINION : Since cinacalcet may not only have an effect on parathyroid P41180 but also on P41180 expressed at bone and renal levels , it can currently only be considered a good alternative to parathyroidectomy in PTH-dependent hypercalcemic conditions when surgical intervention is burdened by a high failure rate or when it can be considered a risky procedure . At present , cinacalcet can not be considered the first choice treatment in asymptomatic primary hyperparathyroidism or in mild-to-moderate forms of familial hypocalciuric hypocalcemia . Vitamin D mediates its action in human colon carcinoma cells in a calcium-sensing receptor-dependent manner : downregulates malignant cell behavior and the expression of thymidylate synthase and survivin and promotes cellular sensitivity to DB00544 . Vitamin D ( VD ) protects against colon carcinogenesis by mechanisms not fully understood . We had earlier reported on the similarity in the biologic action of VD and that of the calcium-sensing receptor ( P41180 ) in human colon carcinoma cells . At the molecular level , the P41180 gene contains 2 VD response elements and VD stimulates the expression of P41180 . In this study , we investigated on the relationship between VD action and P41180 function . We determined and compared the action of VD in human colon carcinoma cells ( P35520 , Moser , Caco-2 and HCT116 ) and their P41180 knocked-down counterparts . VD inhibited cellular proliferation , cellular invasion , and anchorage-independent growth and stimulated the expression of P38936 /Waf1 but not in P41180 knocked-down cells . These results demonstrate , for the first time , that the known tumor-suppressive function of VD requires functional P41180 and knocking down P41180 expression abrogated this function of VD . We recently reported that activation of P41180 in human colon carcinoma cells downregulated the expression of thymidylate synthase ( TS ) and survivin and promoted a significant increase in sensitivity to cytotoxic drugs . We now demonstrate , for the first time , that VD suppressed the expression of TS and survivin , TS and survivin gene transcriptional activities and promoted a cytotoxic response to DB00544 in a P41180 -dependent manner . Ectopic expression of wild-type P41180 in colon carcinoma cells also inhibited the expression of TS and survivin and enhanced cellular sensitivity to DB00544 . VD , however , could no longer enhance cellular sensitivity to DB00544 in cells overexpressing P41180 . DB00502 induces neurotoxicity by the DB01221 receptor downstream signaling pathway , alternative from glutamate excitotoxicity . The DB01221 receptor is believed to be important in a wide range of nervous system functions including neuronal migration , synapse formation , learning and memory . In addition , it is involved in excitotoxic neuronal cell death that occurs in a variety of acute and chronic neurological disorders . Besides of agonist/coagonist sites , other modulator sites , including butyrophenone site may regulate the N-methyl-D-aspartate receptor . It has been shown that haloperidol , an antipsychotic neuroleptic drug , interacts with the Q13224 subunit of DB01221 receptor and inhibits DB01221 response in neuronal cells . We found that DB01221 receptor was co-immunoprecipitated by anti-Ras antibody and this complex , beside NR2 subunit of DB01221 receptor contained haloperidol-binding proteins , P29475 and Ras- P01286 . Furthermore , we have shown that haloperidol induces neurotoxicity of neuronal cells via DB01221 receptor complex , accompanied by dissociation of Ras- P01286 from membranes and activation of c-Jun-kinase . Inclusion of insulin prevented relocalization of Ras- P01286 and subsequent neuronal death . DB00502 -induced dissociation of Ras- P01286 leads to inhibition of membrane-bound form of Ras protein and changes downstream regulators activity that results in the initiation of the apoptotic processes via the mitochondrial way . Our results suggest that haloperidol induces neuronal cell death by the interaction with DB01221 receptor , but through the alternative from glutamate excitotoxicity signaling pathway . DB01221 receptor modulation by the neuropeptide apelin : implications for excitotoxic injury . Excitotoxic neuronal damage via over-activation of the DB01221 receptor has been implicated in many neurodegenerative diseases . In vitro modeling of excitotoxic injury has shown that activation of G-protein coupled receptors ( GPCRs ) counteracts such injury through modulation of neuronal pro-survival pathways and/or DB01221 receptor signaling . We have previously demonstrated that the GPCR P35414 and its endogenous neuropeptide ligand apelin can protect neurons against excitotoxicity , but the mechanism(s) of this neuroprotection remain incompletely understood . We hypothesized that apelin can promote neuronal survival by activating pro-survival signaling as well as inhibiting DB01221 receptor-mediated excitotoxic signaling cascades . Our results demonstrate that ( i ) apelin activates pro-survival signaling via inositol trisphosphate ( IP(3) ) , protein kinase C ( PKC ) , mitogen-activated protein kinase kinase 1/2 ( Q02750 /2 ) , and extracellular signal-regulated kinase-1/2 ( P27361 /2 ) to protect against excitotoxicity , and ( ii ) apelin inhibits excitotoxic signaling by attenuating DB01221 receptor and calpain activity , and by modulating DB01221 receptor subunit Q13224 phosphorylation at serine 1480 . These studies delineate a novel apelinergic signaling pathway that concurrently promotes survival and limits DB01221 receptor-mediated injury to protect neurons against excitotoxicity . Defining apelin-mediated neuroprotection advances our understanding of neuroprotective pathways and will potentially improve our ability to develop therapeutics for excitotoxicity-associated neurodegenerative disorders . Expression of calcium-sensing receptor and characterization of intracellular signaling in human pituitary adenomas . Extracellular Ca(2+)-sensing receptor ( P41180 ) has been recently identified in rat and mouse pituitary and in AtT-20 cells . The aim of the study was to investigate the presence of P41180 in the human pituitary and its signaling pathway . Normal parathyroid biopsies , autoptic normal pituitaries , and seven nonfunctioning and six GH-secreting adenomas were studied . Southern blot analysis of the RT-PCR products from pituitary adenomas indicated that the PCR fragments obtained were products of specific amplification of P41180 messenger ribonucleic acid . Sequence analysis showed nucleotide identity of these products with the available human parathyroid P41180 . By immunoblotting analysis P41180 , was detected in normal and adenomatous pituitary tissues . In all tumors studied , extracellular Ca2+ ( 2.5 mmol/L ) induced a significant increase in intracellular Ca2+ , mainly due to Ca2+ mobilization ( from 82.7+/-11 to 148+/-36 nmol/L ; P < 0.001 ) . Similar results were obtained with the P41180 activators gadolinium and neomycin . Moreover , P41180 activators significantly increased DB02527 levels ; this effect was not mimicked by other agents able to increase intracellular Ca2+ , such as TRH . P41180 agonists did not increase resting GH secretion in any GH-secreting adenomas , but amplified the GH response to P01286 . In this study we first demonstrate P41180 expression in the human pituitary and provides evidence for an additional mechanism by which calcium might regulate pituitary cell function . Enhancement of the P11362 signaling in the P11362 - P08908 heteroreceptor complex in midbrain raphe 5-HT neuron systems . Relevance for neuroplasticity and depression . New findings show existence of P11362 - P08908 heteroreceptor complexes in 5-HT nerve cells of the dorsal and median raphe nuclei of the rat midbrain and hippocampus . Synergistic receptor-receptor interactions in these receptor complexes indicated their enhancing role in hippocampal plasticity . The existence of P11362 - P08908 heteroreceptor complexes also in midbrain raphe 5-HT nerve cells open up the possibility that antidepressant drugs by increasing extracellular 5-HT levels can cause an activation of the P09038 / P11362 mechanism in these nerve cells as well . Therefore , the agonist modulation of the P11362 - P08908 heteroreceptor complexes and their specific role is now determined in rat medullary raphe RN33B cells and in the caudal midline raphe area of the midbrain rich in 5-HT nerve cells . The combined i.c.v. treatment with P09038 and the P08908 agonist 8-OHDPAT synergistically increased P11362 and P27361 /2 phosphorylation in the raphe midline area of the midbrain and in the RN33B cells . Cotreatment with P09038 and the P08908 agonist induced RN33B cell differentiation as seen from development of an increased number and length of extensions per cell and their increased 5-HT immunoreactivity . These signaling and differentiation events were dependent on the receptor interface since they were blocked by incubation with TMV but not by TMII of the P08908 receptor . Taken together , the P08908 autoreceptors by being part of a P11362 - P08908 heteroreceptor complex in the midbrain raphe 5-HT nerve cells appears to have also a trophic role in the central 5-HT neuron systems besides playing a key role in reducing the firing of these neurons . The P38936 codon 31*C- and P14416 codon 313*T-related genotypes/alleles , but not P18887 codon 399 , hOGG1 codon 326 , and P21728 -48 polymorphisms , are correlated with the presence of leiomyoma . OBJECTIVE : To investigate whether the gene polymorphisms for P38936 , X-ray repair cross-complementing group 1 ( P18887 ) , human 8-oxoguanine glycosylase 1 ( hOGG1 ) , and dopamine D1 and D2 receptors ( P21728 , -2 ) are associated with leiomyoma susceptibility . DESIGN : Prospective study . SETTING : Departments of gynecology and genetics in a medical center . PATIENT(S) : Women were divided into two groups : leiomyoma ( n = 120 ) and nonleiomyoma ( n = 112 ) . INTERVENTION(S) : The P38936 codon 31 , P18887 codon 399 , hOGG1 codon 326 , P21728 -48 , and P14416 codon 313 polymorphisms were genotyped by polymerase chain reaction with restriction enzyme digestions ( Blp I , MspI , Fnu4HI , Dde I , and NcoI , respectively ) . MAIN OUTCOME MEASURE(S) : Genotypes and allelic frequencies . RESULT(S) : The P38936 codon 31(*)C- and P14416 codon 313(*)T-related genotypes/alleles were associated with the presence of leiomyomas . The proportions of P38936 (*)CC/CA/AA and P14416 (*)CC/CT/TT in both groups were 27.5/68.3/4.2 % and 12.5/51.7/35.8 % ( leiomyoma ) ; and 14.3/51.8/33.9 % and 33.9/40.2/25.9 % ( nonleiomyoma ) . P18887 , hOGG1 , and P21728 were not correlated with the presence of leiomyomas . P18887 (*)GG/GA/AA , hOGG1(*)TT/TA/AA , and P21728 (*)GG/GA/AA were 54.2/37.5/8.3 % , 36.7/44.2/19.1 % , and 3.3/25.8/70.8 % ( leiomyoma ) ; and 48.2/47.3/4.5 % , 43.6/41/15.4 % , and 3.6/25/71.4 % ( nonleiomyoma ) . CONCLUSION(S) : The P38936 codon 31(*)C- and P14416 codon 313(*)T-related genotypes/alleles were associated with the presence of leiomyoma . P18887 , hOGG1 , and P21728 were not correlated with leiomyoma development . DB00203 enhances neurogenesis and oligodendrogenesis in ischemic brain of middle-aged mouse . Adult neural stem cells give rise to neurons , oligodendrocytes and astrocytes . Aging reduces neural stem cells . Using an inducible nestin-CreER( P24752 )/R26R-yellow fluorescent protein ( YFP ) mouse , we investigated the effect of DB00203 , a phosphodiesterase type 5 ( O76074 ) inhibitor , on nestin lineage neural stem cells and their progeny in the ischemic brain of the middle-aged mouse . We showed that focal cerebral ischemia induced nestin lineage neural stem cells in the subventricular zone ( SVZ ) of the lateral ventricles and nestin expressing NeuN positive neurons and adenomatous polyposis coli ( P25054 ) positive mature oligodendrocytes in the ischemic striatum and corpus callosum in the aged mouse . Treatment of the ischemic middle-aged mouse with DB00203 increased nestin expressing neural stem cells , mature neurons , and oligodendrocytes by 33 , 75 , and 30 % , respectively , in the ischemic brain . These data indicate that DB00203 amplifies nestin expressing neural stem cells and their neuronal and oligodendrocyte progeny in the ischemic brain of the middle-aged mouse . DB01012 : pharmacological and clinical aspects . The calcium sensing receptor ( P41180 ) is expressed in cells secreting calcium-regulating hormones , in cells involved in calcium transport and in many other tissues , with an as yet not completely defined role . In parathyroid cells , the P41180 stimulation inhibits parathyroid hormone ( PTH ) secretion , synthesis and parathyroid cell proliferation . DB01012 belongs to calcimimetic type II compounds that can interact with P41180 , increasing its affinity for calcium . Clinical studies have proved cinacalcet to be effective in reducing calcium and PTH levels in primary hyperparathyroidism and in reducing PTH , calcium and phosphate in patients with secondary hyperparathyroidism owing to chronic renal failure , with a relatively safe profile , the only reported adverse events being hypocalcaemia and gastrointestinal symptoms . However , though calcimimetics do represent a real advancement in the field of the treatment of PTH secretion disturbances , there is a need for clinical trials , which should aim to demonstrate that a better control of biochemical parameters is also matched with better clinical outcomes . Extracellular signal-regulated kinase and the small GTP-binding protein , Rac , contribute to the effects of transforming growth factor-beta1 on gene expression . The kinases and regulatory proteins that convey signals initiated by transforming growth factor-beta ( TGF-beta ) to the nucleus are poorly characterized . To study the role of the extracellular signal-regulated kinase ( P29323 ) pathway in this process , we transiently transfected NIH 3T3 fibroblasts with TGF-beta-responsive luciferase reporter genes and expression vectors designed to interrupt this kinase cascade . Mitogen-activated protein ( Q96HU1 ) kinase phosphatase-1 and a dominant negative Q96HU1 / P29323 kinase 1 mutant reduced stimulation of plasminogen activator inhibitor-1 ( P05121 ) promoter activity by TGF-beta1 from 11.5- to 4-fold and 4.9-fold , respectively . Similar results were observed with the type I collagen promoters . TGF-beta1 increased P27361 activity 4.5-fold at 5 min and 3 . 1-fold at 3 h , while Jun kinase and p38 activity were not affected . Cotransfection of a dominant negative mutant of the small G protein , Rac , but not dominant negative Ras , Cdc42 , or Rho mutants , reduced the effects of TGF-beta1 on the P05121 promoter by approximately half . In support of a role for Rac in signaling by TGF-beta , GTP binding to Rac was increased 3.7-fold following exposure of NIH 3T3 cells to TGF-beta1 for 3 min . These findings indicate that TGF-beta1 modulates gene expression partly through P29323 and Rac in NIH 3T3 cells . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Absence of thyroid transcription factor-1 expression in human parathyroid and pituitary glands . Thyroid transcription factor-1 ( Q15669 -1 ) , a tissue-specific nuclear transcription factor involved in the embryogenesis and differentiation of human thyroid , lung and brain , has been recently identified in other rat tissues , including parafollicular C cells and parathyroid chief cells . Based on this distribution , a possible role for this factor in calcium homeostasis has been suggested . This study investigated the presence of Q15669 -1 transcripts and protein in human tissues expressing the calcium sensing receptor ( P41180 ) . Using a RT-PCR technique , complemented by Southern blot analysis , Q15669 -1 expression was detected in human C cells ( two medullary thyroid carcinomas ) , but not in normal and adenomatous ( four adenomas and three hyperplasia ) parathyroid , and normal and adenomatous ( six adenomas ) pituitary tissues . P41180 was expressed in all samples . The absence of expression was confirmed by Western blot . In contrast to previous studies in the rat , this study demonstrates the absence of Q15669 -1 transcripts in the human adult parathyroid and pituitary glands , although a role for this factor during the ontogeny of these organs can not be excluded . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Impact of clinically relevant mutations on the pharmacoregulation and signaling bias of the calcium-sensing receptor by positive and negative allosteric modulators . DB01012 is predominantly used to treat secondary hyperparathyroidism due to end-stage renal failure , but , more recently , its potential clinical efficacy in treating patients with loss-of-function mutations in the calcium-sensing receptor ( P41180 ) has been recognized . Many clinically relevant P41180 mutations are located in the heptahelical membrane spanning and extracellular loop regions of the receptor , where allosteric modulators are predicted to bind . The aim of the present study was to investigate the impact of such mutations on the pharmacoregulation of the P41180 by the positive and negative allosteric modulators , cinacalcet and P0C0P6 -2143 , respectively . Both cinacalcet and P0C0P6 -2143 effectively rescued mutants whose cell surface expression was substantially impaired , suggesting that both classes of drug can stabilize a receptor conformation that is trafficked more effectively to the cell surface . In addition , functional impairments in almost all mutant CaSRs were rescued by either cinacalcet or P0C0P6 -2143 via restoration of intracellular signaling . There was a significantly greater ability of both compounds to modulate agonist-stimulated intracellular Ca(2+) mobilization than P27361 /2 phosphorylation , indicating that the allosteric modulators engender bias in agonist-stimulated P41180 signaling to different pathways . Three mutations ( G(670)R , P(748)R , and L(773)R ) altered the binding affinity of allosteric modulators to the P41180 , and 3 mutations ( V(817)I , L(773)R , and E(767)K ) altered the cooperativity between the allosteric modulator and Ca(2+)(o) . These findings have important implications for the treatment of diseases associated with P41180 mutations using allosteric P41180 modulators and for analyzing the effects of mutations on the function and pharmacoregulation of the P41180 . Chronic inhibition of cyclic GMP phosphodiesterase 5A prevents and reverses cardiac hypertrophy . Sustained cardiac pressure overload induces hypertrophy and pathological remodeling , frequently leading to heart failure . Genetically engineered hyperstimulation of guanosine 3',5'-cyclic monophosphate ( cGMP ) synthesis counters this response . Here , we show that blocking the intrinsic catabolism of cGMP with an oral phosphodiesterase-5A ( O76074 ) inhibitor ( sildenafil ) suppresses chamber and myocyte hypertrophy , and improves in vivo heart function in mice exposed to chronic pressure overload induced by transverse aortic constriction . DB00203 also reverses pre-established hypertrophy induced by pressure load while restoring chamber function to normal . cGMP catabolism by O76074 increases in pressure-loaded hearts , leading to activation of cGMP-dependent protein kinase with inhibition of O76074 . O76074 inhibition deactivates multiple hypertrophy signaling pathways triggered by pressure load ( the calcineurin/NFAT , phosphoinositide-3 kinase ( PI3K ) /Akt , and P27361 /2 signaling pathways ) . But it does not suppress hypertrophy induced by overexpression of calcineurin in vitro or Akt in vivo , suggesting upstream targeting of these pathways . O76074 inhibition may provide a new treatment strategy for cardiac hypertrophy and remodeling .