interacts_with DB01267?
Agents that block fibronectin fragment-mediated cartilage damage also promote repair . OBJECTIVE AND DESIGN : The objective was to determine if agents that suppress catabolism might also enhance repair of irreversibly damaged cartilage . MATERIAL : Articular cartilage from bovine metacarpophalangeal joints was studied in explant culture . TREATMENT : P02751 fragments or P01583 , which potently cause proteoglycan ( PG ) loss from cartilage , were added to cultures also containing the catabolism-blocking agents : insulin-like growth factor-1 , or DB06151 , or DB00125 - DB00145 - DB00128 - DB00133 peptide , and the effects of these agents on blocking PG loss determined . To test for repair or restoration of PG , cartilage was first damaged , damage agents removed and inhibitory agents added . METHODS : Each mean and SD value for cartilage PG content was determined by assays of papain digests of cartilage from three similar cultures . RESULTS : The agents either partially or fully blocked PG loss and promoted repair . CONCLUSIONS : Normally irreversible cartilage damage was reversed by slowing ongoing catabolic processes during attempted repair . Thus , catabolic inhibitors have reparative potential . Inhibition of human brain and RBC acetylcholinesterase ( P22303 ) by heptylphysostigmine ( HPTL ) . Heptylphysostigmine ( HPTL ) , a derivative of the P22303 inhibitor physostigmine ( PHY ) , is under investigation as a therapeutic agent in Alzheimer 's disease . HPTL is active against human RBC P22303 both in vitro and in vivo . Activity of HPTL against human brain has not been documented . We have developed an in vitro assay system using particulate membrane fractions which permits comparison of inhibition and recovery kinetics of human RBC ( primarily globular dimer ) and brain ( primarily globular tetramer ) membrane-bound forms . Under these conditions the HPTLIC50 is similar for the two forms . RBC P22303 inhibition spontaneously reverses in 24 h , as occurs in vivo . In striking contrast , activity of inhibited brain enzyme does not recover on overnight incubation . DDVP-induced inhibition , but not HPTL inhibition , can be reversed by the oxime DB00733 . Some recovery of HPTL inhibition , but not to the level seen with RBC P22303 , occurs on addition of heat-stable fractions from serum or P04141 . Brain enzyme recovers rapidly from PHY in this system . Responses of brain and RBC P22303 to HPTL indicate that these forms are functionally as well as structurally distinct . Since brain inhibition apparently does not spontaneously reverse like RBC inhibition , peripheral measurements of patient responses should be assessed with caution during treatment with HPTL . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . Partial characterization of a neurotransmitter pathway regulating the in vivo release of prolactin . Nicotinic cholinergic , opiate and serotonergic agonists as well as dopaminergic antagonists induce the release of pituitary prolactin . The purposes of the present studies were to determine if nicotine , morphine and the serotonin1A ( P08908 ) agonist 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) utilize a common synaptic pathway to release prolactin and , if so , to establish the serial order of the receptors involved . We also sought to determine whether the pathway under investigation leads to the secretion of prolactin via a mechanism involving dopamine , the prolactin inhibitory factor . Male rats with indwelling jugular catheters were pretreated with saline , mecamylamine , naltrexone , methysergide or bromocriptine . In the saline-treated animals , administration of nicotine , morphine , 8-OH-DPAT and haloperidol resulted in significant increases in plasma prolactin levels . DB00657 pretreatment prevented the prolactin response to nicotine only . Naltrexone blocked the stimulation of prolactin release by morphine and by nicotine . Methysergide inhibited the effects of 8-OH-DPAT , morphine and nicotine but not haloperidol . DB01200 blocked the prolactin secretion induced by haloperidol as well as by each of the above agonists . Also , in dual-immunocytochemically stained sections , tyrosine hydroxylase-immunoreactive cells and serotonin-immunoreactive processes were detected in close anatomical proximity in the dorsomedial arcuate nucleus . These data indicate that nicotine , morphine and 8-OH-DPAT act to release prolactin via a common synaptic pathway expressing nicotinic cholinergic , opiate , and P08908 receptors at synapses arranged serially in that functional order . Furthermore , the data indicate that the in vivo secretion of prolactin via this pathway may ultimately occur through the inhibition of dopamine release . DB08888 for vitreoretinal diseases . P02751 and laminin are clinically relevant plasmin receptors in the eye . Located at the vitreoretinal interface , they are cleaved by ocriplasmin ( DB05028 , ThromboGenics , Iselin , NJ ) , a novel ophthalmic medication . A series of clinical trials to study ocriplasmin for the treatment of vitreoretinal diseases such as vitreomacular traction , macular hole , and exudative age-related macular degeneration are underway . The results are promising and may impact patient care . Lack of biological relevance of platelet cyclooxygenase-2 dependent thromboxane A2 production . INTRODUCTION : There is emerging evidence of a considerable variability of the impact of aspirin on clinical outcome and laboratory findings . Persistent TxA2 production seems to be the most likely reason . Aim of this study was to determine whether the mechanism responsible for TxA2 persistent production is , at least partially , dependent upon aspirin-insensitive platelet P35354 enzymatic pathway . METHODS AND RESULTS : In 100 consecutive patients , under chronic aspirin anti-platelet treatment ( 100-160 mg/day ) selected on the basis of detectable plasma salicylate levels , serum and DB04557 ( AA ) -induced platelet TxA2 production , immunoblot analysis of platelet P23219 / P35354 expression and P35354 activity were studied . Immunoblot revealed P35354 expression in 46 % patients , in an amount that was markedly lower than P23219 . In 10 P35354 positive patients with TxA2 levels over the median , AA-induced TxA2 production performed in vitro in the presence of the P35354 inhibitor CAY10404 and aspirin demonstrated that P35354 dependent TxA2 production is less than 2 % . CONCLUSION : Our data demonstrate that the inter-individual variability of platelet sensitivity to aspirin is due to a reduced efficacy of aspirin on platelet P23219 despite ascertained patient compliance . We suggest that serum TxA2 assay might be performed in future clinical studies to improve our knowledge on the residual TxA2 production in aspirin-treated patients . Critical role of Q08499 in beta2-adrenoceptor-dependent DB02527 signaling in mouse embryonic fibroblasts . One of the defining properties of beta2-adrenergic receptor ( beta(2)AR ) signaling is the transient and rapidly reversed accumulation of DB02527 . Here we have investigated the contribution of different DB05876 proteins to the generation of this transient response . To this aim , mouse embryonic fibroblasts deficient in P27815 , Q07343 , or Q08499 were generated , and the regulation of PDE activity , the accumulation of DB02527 , and CREB phosphorylation in response to isoproterenol were monitored . Ablation of Q08499 , but not P27815 or Q07343 , had a major effect on the beta-agonist-induced PDE activation , with only a minimal increase in PDE activity being retained in Q08499 knock-out ( KO ) cells . Accumulation of DB02527 was markedly enhanced , and the kinetics of DB02527 accumulation were altered in their properties in PDE4DKO but not PDE4BKO cells . Modest effects were observed in PDE4AKO mouse embryonic fibroblasts . The return to basal levels of both DB02527 accumulation and CREB phosphorylation was greatly delayed in the PDE4DKO cells , suggesting that Q08499 is critical for dissipation of the beta2AR stimulus . This effect of Q08499 ablation was in large part due to inactivation of a negative feedback mechanism consisting of the PKA-mediated activation of Q08499 in response to elevated DB02527 levels , as indicated by experiments using the DB02527 -dependent protein kinase inhibitors H89 and PKI . Finally , Q08499 ablation affected the kinetics of beta2AR desensitization as well as the interaction of the receptor with Galphai . These findings demonstrate that Q08499 plays a major role in shaping the beta2AR signal . Human chromosome 3 and pig chromosome 13 show complete synteny conservation but extensive gene-order differences . A comparative map of human chromosome 3 ( HSA 3 ) and pig chromosome 13 ( SSC 13 ) was constructed using physically assigned pig sequence-tagged sites ( STSs ) . Pig STSs representing 11 HSA 3 genes , including v- P04049 murine leukemia viral oncogene homolog 1 ( P04049 ) , retinoic acid beta receptor ( P10826 ) , cholecystokinin ( CCK ) , pituitary transcription factor 1 ( P28069 ) , ceruloplasmin ( CP ) , guanine nucleotide binding protein , alpha-inhibiting polypeptide 2 ( P04899 ) , sucrase-isomaltase ( SI ) , rhodopsin ( P08100 ) , dopamine receptor D3 ( P35462 ) , growth-associated protein 43 ( P17677 ) , and somatostatin ( P61278 ) , were developed . Ten pig STSs were regionally mapped using a somatic cell hybrid panel ( SCHP ) to SSC 13 with 80-100 % concordance . Large-insert probes were obtained by screening a pig yeast artificial chromosome ( YAC ) library with primers for each STS . Several YACs were identified for P35462 , P17677 , P28069 , P08100 , SI , and P61278 for fluorescence in situ hybridization ( Q5TCZ1 ) mapping . Single gene and bi-color Q5TCZ1 with each pairwise combination were used to further define the gene order on SSC 13 . While these data confirm chromosome painting results showing that HSA 3 probes hybridize to a major portion of SSC 13 , they also demonstrate extensive gene-order differences between man and pig within this large conserved synteny group . Interestingly , several conserved chromosomal regions have been detected between pig and mouse that are not conserved between man and mouse , suggesting that the SSC 13 gene arrangement may be the closest to that of the ancestral eutherian chromosome . Transcriptional modulation of monoaminergic neurotransmission genes by the histone deacetylase inhibitor trichostatin A in neuroblastoma cells . Histone deacetylase inhibitors are promising anti-tumor agents partly due to their ability to disrupt the hypoxic signaling pathway in human malignancies . However , little is known about any effects of these drugs on the central nervous system . The aim of the present study was to analyze the effects of trichostatin A ( P32119 ) -- a broad-spectrum histone deacetylase inhibitor -- on the transcriptional regulation of several genes involved in dopamine- and serotonergic neurotransmission . To this end , short-term parallel cultures of SK-NF-I neuroblastoma cells were treated with P32119 either alone or in combination with hypoxia , and mRNA levels of dopamine receptor D3 ( P35462 ) and D4 ( P21917 ) , dopamine transporter ( Q01959 ) , dopamine hydroxylase ( P09172 ) , dopamine receptor regulating factor ( DRRF ) , catechol-O-methyltransferase ( P21964 ) , serotonin receptor 1A ( P08908 ) , monoamino oxidase A ( P21397 ) , serotonin transporter ( P31645 ) and tryptophan hydroxylase 2 ( Q8IWU9 ) were determined by quantitative PCR . We found that P32119 did not antagonize the hypoxia-induced activation of D3 and D4 dopamine receptor genes , implying that induction of these genes is not mediated directly by hypoxia inducible factor-1alpha . On the other hand , P32119 dramatically upregulated the expression of Q01959 and P31645 ( 45-fold and 15-fold , respectively ) , while transcript levels of P21397 and P21964 were significantly reduced ( by 70 % and by more than 90 % , respectively ) . Induction of Q01959 protein expression was detected by western blotting . These results suggest that inhibition of histone deacetylases might help restore presynaptic monoamine pools via suppression of catecholamine breakdown and facilitation of monoamine reuptake in neurons . Perfusion-independent effect of bradykinin and fosinoprilate on glucose transport in Langendorff rat hearts . P12821 ( P12821 ) inhibitor-stimulated glucose metabolism and perfusion in muscle tissue seem to be , at least in part , mediated by kinins . However , the relative contribution of direct metabolic or secondary hemodynamically induced effects is unclear . It was the aim of this study to characterize the effects of P12821 inhibition and bradykinin on glucose transport while changes in cardiocoronary function that might influence glucose transport were minimized . Hearts from Wistar rats were perfused by a Langendorff preparation and a set of functional parameters were simultaneously measured . Bradykinin ( 10[-11] M ) and fosinoprilate ( 10[-7] M ) were administered at concentrations that did not affect coronary flow . P01308 was employed as reference at half-maximal concentration . The nonmetabolizable glucose analog 3-O-[14C]methyl-D-glucose and the nontransportable tracer L-[3H]glucose were coperfused for the calculation of glucose transport . Using a 2-compartment mathematical model we found that the glucose transport rate , which was doubled with insulin , was increased almost 3-fold by either bradykinin or fosinoprilate . In the presence of the P30411 antagonist DB06196 ( D- DB00125 [Hyp3,Thi5,D-Tic7,Oic8]-bradykinin ; icatibant ) , the effect of both agents was completely abolished . Both agents also induced minor changes in contractility/relaxation parameters that again were completely neutralized with icatibant . A perfusion-independent but B2-kinin receptor-dependent stimulating effect on glucose transport by either bradykinin or fosinoprilate is concluded . This effect could , in analogy to insulin be due to increased glucose transporter translocation , increased endothelium-derived nitric oxide formation , or -- despite constant coronary flow conditions -- secondary to altered cardiac function . Characterization of the interaction of ingenol 3-angelate with protein kinase C . DB05013 ( I3A ) is one of the active ingredients in Euphorbia peplus , which has been used in traditional medicine . Here , we report the initial characterization of I3A as a protein kinase C ( PKC ) ligand . I3A bound to P17252 in the presence of phosphatidylserine with high affinity ; however , under these assay conditions , little PKC isoform selectivity was observed . PKC isoforms did show different sensitivity and selectivity for down-regulation by I3A and phorbol 12-myristate 13-acetate ( PMA ) in WEHI-231 , Q9BPY8 -92 , and Colo-205 cells . In all of the three cell types , I3A inhibited cell proliferation with somewhat lower potency than did PMA . In intact CHO- P04264 cells , I3A was able to translocate different green fluorescent protein-tagged PKC isoforms , visualized by confocal microscopy , with equal or higher potency than PMA . PKC-delta in particular showed a different pattern of translocation in response to I3A and PMA . I3A induced a higher level of secretion of the inflammatory cytokine interleukin 6 compared with PMA in the WEHI-231 cells and displayed a marked biphasic dose-response curve for the induction . I3A was unable to cause the same extent of association of the C1b domain of PKC-delta with lipids , compared with PMA or the physiological regulator diacylglycerol , and was able to partially block the association induced by these agents , measured by surface plasmon resonance . The in vitro kinase activity of P17252 induced by I3A was lower than that induced by PMA . The novel pattern of behavior of I3A makes it of great interest for further evaluation . Nonlinkage of bipolar illness to tyrosine hydroxylase , tyrosinase , and D2 and D4 dopamine receptor genes on chromosome 11 . OBJECTIVE : Previous linkage and allelic association studies using DNA polymorphisms , cosegregation of cytogenetic abnormalities with psychiatric illness , and assignment of genes involved in neutotransmitter metabolism suggested that chromosome 11 may harbor a gene predisposing to bipolar illness . The authors examined linkage in the families of 14 probands with bipolar illness , with the candidate genes tyrosine hydroxylase ( TH ) , D4 dopamine receptor ( P21917 ) at 11p15 , tyrosinase ( P14679 ) at 11q14-q21 , and D2 dopamine receptor ( P14416 ) at 11q22-q23 , as well as with the c-Harvey-ras oncogene ( P01112 ) and insulin gene ( P01308 ) , both located at 11p15 , a region that previously showed linkage to bipolar illness . METHOD : The genetic data were analyzed with both lod score analysis ( parametric ) and affected-sib-pair analysis ( nonparametric ) ; both narrow and broad definitions of the clinical phenotype were used . Further influences of diagnostic uncertainties were accounted for by using diagnostic probability classes weighing the stability of each phenotype . RESULTS : Two-point linkage results excluded close linkage of bipolar illness to each candidate gene ; negative results were also obtained when the narrow definition of the clinical phenotype was used . Moreover , multipoint linkage analysis of P01112 and P01308 excluded the 11p15 region encompassing both P21917 and TH . In agreement with the negative linkage results , affected-sib-pair analysis did not show preferential sharing of marker alleles at any of the candidate genes . CONCLUSIONS : The negative results obtained under different genetic models exclude a frequent role for P21917 , TH , P14679 , and P14416 in the pathogenesis of bipolar illness . Functional analysis of the human D2 dopamine receptor missense variants . The human dopamine D2 receptor gene ( P14416 ) has three polymorphic variants that predict the amino acid substitutions Val96 --> Ala , Pro310 --> DB00133 , and Ser311 --> DB00151 in the receptor protein . We have investigated the ligand binding and signal transduction properties of these human D2 receptor variants by stably expressing them in cultured mammalian cells . The Cys311 and Ser310 variants of the human D2 receptor , which involve substitutions located in the third cytoplasmic loop , were markedly less effective in inhibiting DB02527 synthesis than the most prevalent form ( Pro310 , Ser311 ) . Despite this difference , the Cys311 and Ser310 variants couple to G proteins in CHO- P04264 ( Chinese hamster ovary ) cells . The impairment of the Cys311 and Ser310 variants to inhibit DB02527 levels thus appears to result from a reduced ability of those variant receptors to activate the appropriate Gi-like protein . The demonstration of substantial functional differences between P14416 gene variants found in the human population might have important pharmacological implications given the widespread use of D2 receptor blocking drugs in the treatment of schizophrenia and other psychiatric disorders . ZNF804a regulates expression of the schizophrenia-associated genes Q9NQE7 , P21964 , Q07343 , and P14416 . ZNF804a was identified by a genome-wide association study ( GWAS ) in which a single nucleotide polymorphism ( SNP rs1344706 ) in ZNF804a reached genome-wide statistical significance for association with a combined diagnosis of schizophrenia ( SZ ) and bipolar disorder . Although the molecular function of ZNF804a is unknown , the amino acid sequence is predicted to contain a C2H2-type zinc-finger domain and suggests ZNF804a plays a role in DNA binding and transcription . Here , we confirm that ZNF804a directly contributes to transcriptional control by regulating the expression of several SZ associated genes and directly interacts with chromatin proximal to the promoter regions of Q9NQE7 and P21964 , the two genes we find upregulated by ZNF804a . Using immunochemistry we establish that ZNF804a is localized to the nucleus of rat neural progenitor cells in culture and in vivo . We demonstrate that expression of ZNF804a results in a significant increase in transcript levels of Q9NQE7 and P21964 , relative to GFP transfected controls , and a statistically significant decrease in transcript levels of Q07343 and P14416 . Furthermore , we show using chromatin immunoprecipitation assays ( ChIP ) that both epitope-tagged and endogenous ZNF804a directly interacts with the promoter regions of Q9NQE7 and P21964 , suggesting a direct upregulation of transcription by ZNF804a on the expression of these genes . These results are the first to confirm that ZNF804a regulates transcription levels of four SZ associated genes , and binds to chromatin proximal to promoters of two SZ genes . These results suggest a model where ZNF804a may modulate a transcriptional network of SZ associated genes . Characterization of the pattern of the nongenomic signaling pathway through which TCDD-induces early inflammatory responses in U937 human macrophages . 2,3,7,8-Tetrachlorodibenzo(p)dioxin ( TCDD ) has been known to induce inflammatory signaling in a number of cell types and tissues . We found that in U937 macrophages TCDD causes rapid activation of cytosolic phospholipase A2 ( P47712 ) within 30min as judged by the increase in the serine 505 phosphorylated form of P47712 protein and the increased cellular release of free arachidonic acid . This initial action of TCDD is accompanied with the up-regulation of an important inflammation marker , P35354 mRNA expression within 1h , and by 3h , several other markers become up-regulated . These effects appear to be dependent on the initial increase in the intracellular concentration of Ca(2+) , and activation of P47712 and P35354 . A comparative study among three different human cell lines showed that activation of P35354 within 1h of action of TCDD is a common feature exhibited by all cell lines . On the other hand , the U937 macrophage line appears to be unique among them with respect to its ability to activate P01375 and P10145 mRNA expressions , and not requiring Src kinase in propagating the initial signaling of P47712 . Based on the rapidity of activation of P47712 and P35354 , which occurs within 1h of cell exposure to TCDD , when no change in mRNA expression of P04798 has been observed , it is apparent that this unique action of TCDD is carried out through a distinct " nongenomic " pathway which , is clearly discernable from the classical , " genomic " action pathway of the P35869 by not requiring the participation of P27540 . Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples . An analysis of the effects of retinoic acid and other retinoids on the development of adrenergic cells from the avian neural crest . In the present work , we have investigated the role of all-trans-retinoic acid ( all-trans RA ) , and several other natural and synthetic retinoids , in the development of adrenergic cells in quail neural crest cultures . Dose response studies using all-trans RA and 13-cis RA revealed a dose-dependent increase in the number of adrenergic cells in neural crest cultures . Similar dose response studies using RA isomers and other natural retinoids did not result in the same increases . In order to determine the receptor mediating the effects of all-trans RA in the neural crest , we tested several synthetic analogs which specifically bind to a particular RA receptor ( RAR ) subtype . We found that the compound AM 580 , which activates the P10276 , produced an increase in adrenergic cells similar to that seen with all-trans RA . The compound DB02877 , which activates all RAR subtypes , also resulted in an increase in adrenergic cells . We conclude that the increase in adrenergic cells seen with all-trans RA is mediated by P10276 and possibly P10826 . To further define the actions of all-trans RA on the neural crest we incubated cultures with 5-bromo-2'-deoxyuridine ( BrdU ) to determine whether all-trans RA could affect the rate of proliferation . The results show that while all-trans RA did not increase the fraction of cells incorporating BrdU into their nuclei at early time points ( 24 h ) , it did increase BrdU incorporation by tyrosine hydroxylase ( TH ) positive cells at 5 days in culture . These findings demonstrate that the increase in adrenergic cells seen with all-trans RA in neural crest cultures is likely due to an increase in the proliferation of cells already expressing TH . Quantification of raf antisense oligonucleotide ( rafAON ) in biological matrices by LC-MS/MS to support pharmacokinetics of a liposome-entrapped rafAON formulation . An LC-MS/MS method was developed to quantify an antisense oligonucleotide against P04049 expression ( rafAON ) in monkey and mouse plasma and in mouse tissue homogenates from animals dosed with a liposome-entrapped rafAON easy-to-use formulation ( DB04973 -ETU ) intended for use as an antineoplastic agent . RafAON was extracted from mouse and monkey plasma using solid-phase extraction . Tissues were homogenized and sample cleanup was achieved by protein precipitation . RafAON and the internal standard ( IS ) were separated on a Hamilton PRP-1 column and quantified by tandem mass spectrometry using an electrospray source in negative ion mode . The total run time was 4.0 min . The peak areas of two rafAON transitions were summed and plotted against the peak area of an IS transition to generate the standard curve . In monkey plasma the linear range was 50-10,000 ng/mL , and in mouse plasma it was 25-5000 ng/mL . The lower limit of quantification was 500 ng/mL ( 10 microg/g tissue ) in heart , kidney , liver , lung and spleen homogenates , and the standard curve was linear up to 10,000 ng/mL . Accuracy , precision and stability were evaluated and found to be acceptable in all three matrices . The assay was used to support pharmacokinetics and tissue distribution studies of DB04973 -ETU in mice and monkeys .