interacts_with DB01656?
Q9NRI5 -binding proteins in neural development , signalling and schizophrenia . In the decade since Disrupted in Schizophrenia 1 ( Q9NRI5 ) was first identified it has become one of the most convincing risk genes for major mental illness . As a multi-functional scaffold protein , Q9NRI5 has multiple identified protein interaction partners that highlight pathologically relevant molecular pathways with potential for pharmaceutical intervention . Amongst these are proteins involved in neuronal migration ( e.g. P05067 , Dixdc1 , P43034 , Q9NXR1 , Q9GZM8 ) , neural progenitor proliferation ( GSK3β ) , neurosignalling ( Q3V6T2 , GSK3β , DB05876 ) and synaptic function ( Kal7 , Q9UKE5 ) . Furthermore , emerging evidence of genetic association ( Q9GZM8 , PCM1 , Q07343 ) and copy number variation ( Q9NXR1 ) implicate several Q9NRI5 -binding partners as risk factors for schizophrenia in their own right . Thus , a picture begins to emerge of Q9NRI5 as a key hub for multiple critical developmental pathways within the brain , disruption of which can lead to a variety of psychiatric illness phenotypes . Phosphodiesterase 4B mediates extracellular signal-regulated kinase-dependent up-regulation of mucin P98088 protein by Streptococcus pneumoniae by inhibiting DB02527 -protein kinase A-dependent P28562 phosphatase pathway . Otitis media ( OM ) is the most common childhood bacterial infection and the major cause of conductive hearing loss in children . Mucus overproduction is a hallmark of OM . Streptococcus pneumoniae is the most common gram-positive bacterial pathogen causing OM . Among many mucin genes , P98088 has been found to be greatly up-regulated in the middle ear mucosa of human patients with OM . We previously reported that S. pneumoniae up-regulates P98088 expression in a MAPK P29323 -dependent manner . We also found that MAPK phosphatase-1 ( P28562 ) negatively regulates S. pneumoniae-induced P29323 -dependent P98088 up-regulation . Therapeutic strategies for up-regulating the expression of negative regulators such as P28562 may have significant therapeutic potential for treating mucus overproduction in OM . However , the underlying molecular mechanism by which P28562 expression is negatively regulated during S. pneumoniae infection is unknown . In this study we show that phosphodiesterase 4B ( Q07343 ) mediates S. pneumoniae-induced P98088 up-regulation by inhibiting the expression of a negative regulator P28562 , which in turn leads to enhanced MAPK P29323 activation and subsequent up-regulation of P98088 . Q07343 inhibits P28562 expression in a DB02527 -PKA-dependent manner . DB05876 -specific inhibitor rolipram inhibits S. pneumoniae-induced P98088 up-regulation both in vitro and in vivo . Moreover , we show that Q07343 plays a critical role in P98088 induction . Finally , topical and post-infection administration of rolipram into the middle ear potently inhibited S. pneumoniae-induced P98088 up-regulation . Collectively , these data demonstrate that Q07343 mediates P29323 -dependent up-regulation of mucin P98088 by S. pneumoniae by inhibiting DB02527 -PKA-dependent P28562 pathway . This study may lead to novel therapeutic strategy for inhibiting mucus overproduction . Bronchodilators modulate inflammation in chronic obstructive pulmonary disease subjects . Chronic obstructive pulmonary disease ( P48444 ) is characterized by neutrophilic airway inflammation and oxidative stress . Leukotriene B₄ ( LTB₄ ) , a potent proinflammatory mediator , is synthesized by P09917 ( P09917 ) , which is activated by the presence of lipid hydroperoxides resulting from oxidative stress on biological membranes . We proposed to evaluate the effect of a four week treatment with two different bronchodilators of common practice in P48444 treatment , on the production of reactive oxygen species ( ROS ) , in particular superoxide anions , and of LTB₄ by peripheral blood neutrophils obtained from P48444 subjects . 24 subjects among the P48444 outpatients were enrolled , and randomized to receive either formoterol ( 12 μg bid ) or tiotropium ( 18 μg od ) . Peripheral blood neutrophils were obtained at the start and at the end of the treatment , and production of superoxide anions and of LTB₄ were evaluated as previously published . The results obtained showed a decrease in the unstimulated production of superoxide by isolated neutrophils in both groups , but tiotropium only was effective in modulating the production of LTB₄ , while formoterol caused an increased production of superoxide in response to fMLP , when compared to values obtained before treatment . In conclusion , tiotropium showed a better antiinflammatory activity profile when compared to formoterol in a clinical setting , reducing superoxide and LTB₄ production by peripheral neutrophils obtained from P48444 subjects . Biochemical properties of V91G calmodulin : A calmodulin point mutation that deregulates muscle contraction in Drosophila . A mutation ( Cam7 ) to the single endogenous calmodulin gene of Drosophila generates a mutant protein with valine 91 changed to glycine ( V91G D- P62158 ) . This mutation produces a unique pupal lethal phenotype distinct from that of a null mutation . Genetic studies indicate that the phenotype reflects deregulation of calcium fluxes within the larval muscles , leading to hypercontraction followed by muscle failure . We investigated the biochemical properties of V91G D- P62158 . The effects of the mutation on free P62158 are minor : DB01373 binding , and overall secondary and tertiary structure are indistinguishable from those of wild type . A slight destabilization of the C-terminal domain is detectable in the calcium-free ( apo- ) form , and the calcium-bound ( holo- ) form has a somewhat lower surface hydrophobicity . These findings reinforce the indications from the in vivo work that interaction with a specific P62158 target(s) underlies the mutant defects . In particular , defective regulation of ryanodine receptor ( RyR ) channels was indicated by genetic interaction analysis . Studies described here establish that the putative P62158 binding region of the Drosophila RyR ( D-RyR ) binds wild-type D- P62158 comparably to the equivalent P62158 -RyR interactions seen for the mammalian skeletal muscle RyR channel isoform ( P21817 ) . The V91G mutation weakens the interaction of both apo- and holo-D- P62158 with this binding region , and decreases the enhancement of the calcium-binding affinity of P62158 that is detectable in the presence of the RyR target peptide . The predicted functional consequences of these changes are consonant with the in vivo phenotype , and indicate that D-RyR is one , if not the major , target affected by the V91G mutation in P62158 . Cross-talk between PKA-Cβ and p65 mediates synergistic induction of Q07343 by roflumilast and NTHi . Phosphodiesterase 4B ( Q07343 ) plays a key role in regulating inflammation . DB01656 , a phosphodiesterase (PDE)4-selective inhibitor , has recently been approved for treating severe chronic obstructive pulmonary disease ( P48444 ) patients with exacerbation . However , there is also clinical evidence suggesting the development of tachyphylaxis or tolerance on repeated dosing of roflumilast and the possible contribution of Q07343 up-regulation , which could be counterproductive for suppressing inflammation . Thus , understanding how Q07343 is up-regulated in the context of the complex pathogenesis and medications of P48444 may help improve the efficacy and possibly ameliorate the tolerance of roflumilast . Here we show that roflumilast synergizes with nontypeable Haemophilus influenzae ( NTHi ) , a major bacterial cause of P48444 exacerbation , to up-regulate PDE4B2 expression in human airway epithelial cells in vitro and in vivo . Up-regulated PDE4B2 contributes to the induction of certain important chemokines in both enzymatic activity-dependent and activity-independent manners . We also found that protein kinase A catalytic subunit β ( PKA-Cβ ) and nuclear factor-κB ( NF-κB ) p65 subunit were required for the synergistic induction of PDE4B2 . PKA-Cβ phosphorylates p65 in a DB02527 -dependent manner . Moreover , Ser276 of p65 is critical for mediating the PKA-Cβ-induced p65 phosphorylation and the synergistic induction of PDE4B2 . Collectively , our data unveil a previously unidentified mechanism underlying synergistic up-regulation of PDE4B2 via a cross-talk between PKA-Cβ and p65 and may help develop new therapeutic strategies to improve the efficacy of DB05876 inhibitor . Q13639 receptor agonists increase sAPPalpha levels in the cortex and hippocampus of male C57BL/6j mice . BACKGROUND AND PURPOSE : A strategy to treat Alzheimer 's disease ( AD ) is to increase the soluble form of amyloid precursor protein ( sAPPalpha ) , a promnesic protein , in the brain . Because strong evidence supports beneficial effects of 5-hydroxytryptamine 5-HT(4) receptor agonists in memory and learning , we investigated the role of 5-HT(4) receptors on P05067 processing in 8 weeks-old male C57BL/6j mice . EXPERIMENTAL APPROACH : Mice were given , subcutaneously , prucalopride or ML 10302 ( s.c. ) , two highly selective 5-HT(4) receptor agonists and , up to 240 min later , the hippocampus and cortex were analysed by Western blot for sAPPalpha determination . KEY RESULTS : DB06480 ( 5 or 10 mg kg(-1) ) significantly increased sAPPalpha levels in the hippocampus and cortex , but did not modify the expression level of P05067 mRNA as detected by quantitative RT-PCR . A selective 5-HT(4) receptor antagonist , GR125487 ( 1 mg kg(-1) , s.c. ) inhibited prucalopride induced- increase in sAPPalpha levels . In addition , levels of sAPPalpha were increased by ML10302 only at 20 mg kg(-1) and was limited to the cortex . Also , prucalopride increased sAPPalpha levels in the cortex of a transgenic mouse model of AD , expressing the London mutation of P05067 . Furthermore , the combined injection of a selective acetylcholinesterase inhibitor , donepezil and prucalopride induced a synergic increase in sAPPalpha levels in the cortex and hippocampus . CONCLUSIONS AND IMPLICATIONS : Our results demonstrate that the 5-HT(4) receptor plays a key role in the non-amyloidogenic pathway of P05067 metabolism in vivo and give support to the beneficial use of 5-HT(4) agonists for AD treatment . Identification of the fused bicyclic 4-amino-2-phenylpyrimidine derivatives as novel and potent DB05876 inhibitors . 2-Phenyl-4-piperidinyl-6,7-dihydrothieno[3,4-d]pyrimidine derivative ( 2 ) was found to be a new DB05876 inhibitor with moderate Q07343 activity ( IC50=150 nM ) . A number of derivatives with a variety of 4-amino substituents and fused bicyclic pyrimidines were synthesized . Among these , 5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine derivative ( 18 ) showed potent Q07343 inhibitory activity ( IC50=25 nM ) . Finally , N-propylacetamide derivative ( 31b ) was determined as a potent inhibitor for both Q07343 ( IC50=7.5 nM ) and P01375 -α production in mouse splenocytes ( IC50=9.8 nM ) and showed good in vivo anti-inflammatory activity in the LPS-induced lung inflammation model in mice ( ID50=18 mg/kg ) . The binding mode of the new inhibitor ( 31e ) in the catalytic site of Q07343 is presented based on an X-ray crystal structure of the ligand-enzyme complex . PEGylation of growth hormone-releasing hormone ( P01286 ) analogues . Synthetically produced GRF1-29 ( DB00010 ) has an amino acid composition identical to the N-terminal 29 amino acids sequence of the natural hypothalamic GHRH1-44 ( Figure 1 ) . It maintains bioactivity in vitro and is almost equally effective in eliciting secretion of endogenous growth hormone in vivo . The main drawbacks associated with the pharmaceutical use of hGRF1-29 relate to its short half-life in plasma , about 10-20 min in humans , which is caused mostly by renal ultrafiltration and enzymatic degradation at the N terminus . PEGylation has been considered as one valid approach to obtain more stable forms of the peptide , with a longer in vivo half-life and ultimately with increased pharmacodynamic response along the somatotropic axis ( endogenous GH , DB01277 levels ) . Different PEGylated P01286 conjugates were obtained and their bioactivity was tested in vitro and in vivo by monitoring endogenous growth hormone ( GH ) serum levels after intravenous ( i.v. ) injection in rats , and intravenous and subcutaneous ( s.c. ) injection in pigs . It was found that P01286 -PEG conjugates are able to bind and activate the human Q02643 , although with different potency . The effect of PEG molecular weight , number of PEG chains bound and position of PEGylation site on P01286 activity were investigated . Mono-PEGylated isomers with a PEG5000 polymer chain linked to Lys 12 or Lys 21 residues , showed high biological activity in vitro , which is similar to that of hGRF1-29 , and a higher pharmacodynamic response as compared to unmodified P01286 molecule . Development of pH-responsive fluorescent false neurotransmitters . We introduce pH-responsive fluorescent false neurotransmitters ( pH-responsive FFNs ) as novel probes that act as vesicular monoamine transporter ( VMAT ) substrates and ratiometric fluorescent pH sensors . The development of these agents was achieved by systematic molecular design that integrated several structural elements , including the aminoethyl group ( VMAT recognition ) , halogenated hydroxy-coumarin core ( ratiometric optical pH sensing in the desired pH range ) , and N- or C-alkylation ( modulation of lipophilicity ) . Of 14 compounds that were synthesized , the probe Mini202 was selected based on the highest uptake in Q05940 -transfected P29320 cells and desirable optical properties . Using Mini202 , we measured the pH of catecholamine secretory vesicles in PC-12 cells ( pH approximately 5.9 ) via two-photon fluorescence microscopy . Incubation with methamphetamine led to an increase in vesicular pH ( pH approximately 6.4 ) , consistent with a proposed mechanism of action of this psychostimulant , and eventually to redistribution of vesicular content ( including Mini202 ) from vesicles to cytoplasm . Mini202 is sufficiently bright , photostable , and suitable for two-photon microscopy . This probe will enable fundamental neuroscience and neuroendocrine research as well as drug screening efforts . Genetic analysis of 56 polymorphisms in 17 genes involved in methionine metabolism in patients with abdominal aortic aneurysm . BACKGROUND : Previous studies suggested an association between abdominal aortic aneurysm ( AAA ) and hyperhomocysteinaemia , a complex trait determined by genetic and environmental factors . Our hypothesis was that polymorphisms in genes directly or indirectly involved in methionine metabolism may contribute to AAA susceptibility . METHOD : We studied 56 polymorphisms in P42898 , Q99707 , Q9UBK8 , P35520 , P11586 , P41440 , P40261 , P20062 , P23526 , Q93088 , Q9H2M3 , Q04609 , P04818 , Q7L5Y1 , P34896 , P27169 , Q15165 genes according to their demonstrated/putative function , localisation in promoter or regulatory and coding regions and/or heterozygosity values > 0.300 . Polymorphisms were evaluated by using a primer extension based microarray technology in 423 AAA patients and 423 matched controls . RESULTS : All polymorphisms resulted in Hardy-Weinberg equilibrium in patients and controls . At the multiple logistic regression analysis adjusted for traditional cardiovascular risk factors ( sex , age , hypertension , smoking habit , dyslipidaemia , diabetes ) and chronic obstructive pulmonary disease ( P48444 ) , rs8003379 P11586 ( odds ratio ( OR ) 0.41 , 95 % confidence interval ( CI ) 0.26 to 0.65 ) and rs326118 Q9UBK8 ( OR 0.47 , 95 % CI 0.29 to 0.77 ) polymorphisms resulted in independent susceptibility factor for AAA . CONCLUSIONS : After haplotype reconstruction , logistic regression analyses adjusted for traditional risk factors and P48444 showed a significant association among AAA and P23526 , Q04609 , P11586 , Q99707 , P40261 , P27169 and P04818 haplotypes . Our findings offer new insights into the pathogenesis of AAA . Inhibition of angiogenic and non-angiogenic targets by sorafenib in renal cell carcinoma ( RCC ) in a RCC xenograft model . BACKGROUND : It is widely recognised that sorafenib inhibits a range of molecular targets in renal cell carcinoma ( RCC ) . In this study , we aim to use patient-derived RCC xenografts to delineate the angiogenic and non-angiogenic molecular targets of sorafenib therapy for advanced RCC ( aRCC ) . METHODS : We successfully generated three patient RCC-derived xenografts in severe combined immunodeficient mice , consisting of three different RCC histological subtypes : conventional clear cell , poorly differentiated clear cell RCC with sarcomatoid changes , and papillary RCC . This study also used clear cell RCC cells ( 786-0/EV ) harbouring mutant P40337 to investigate the clonogenic survival of cells transfected with survivin sense and antisense oligonucleotides . RESULTS : All three xenografts retain their original histological characteristics . We reported that sorafenib inhibited all three RCC xenograft lines regardless of histological subtypes in a dose-dependant manner . DB00398 -induced growth suppression was associated with not only inhibition of angiogenic targets p- P09619 -β , p- P35968 , and their downstream signalling pathways p-Akt and p- P29323 , cell cycle , and anti-apoptotic proteins that include cyclin D1 , cyclin B1 , and survivin but also upregulation of proapoptotic Bim . Survivin knockdown by survivin-specific antisense-oligonucleotides inhibited colony formation and induced cell death in clear cell RCC cells . CONCLUSION : This study has shed light on the molecular mechanisms of sorafenib in RCC . Inhibition of non-angiogenic molecules by sorafenib could contribute in part to its anti-tumour activities observed in vivo , in addition to its anti-angiogenic effects . Serotonin neurotoxins -- past and present . Autoxidation pathways and redox reactions of dihydroxytryptamines ( 5,6- and 5,7- DB02901 ) and of 6-hydroxydopamine ( 6-OH-DA ) are illustrated , and their potential role in aminergic neurotoxicity is discussed . It is proposed that certain aspects of the cytotoxicity of 6-OH-DA and of the DHTs , namely redox cycling of their quinone- and quinoneimine-intermediates as a source of free radicals , may also apply to quinoidal reactive intermediates and to glutathionyl- or cysteinyl conjugates ( " thioether adducts " ) of o-dihydroxylated ( catechol-like ) metabolites of certain substituted amphetamines ( of methylenedioxymethamphetamine ( DB01454 ) and of methylenedioxyamphetamine ( MDA ) ) . Despite similarities in their primary interaction with the plasmalemmal ( serotonergic transporter/dopamine transporter , P31645 / Q01959 ) and vesicular monoamine transporters ( Q05940 ) , DB01454 and fenfluramine ( N-ethyl-meta-trifluoromethamphetamine , Fen ) differ substantially in many aspects of their metabolism , pharmacokinetics , pharmacology , and neurotoxicology profile ; the consequences of these differences for neuronal response patterns and long-term survival prospects are not yet fully understood . However , sustained hyperthermia appears to be a critical factor in these differences . Methodological requirements for adequate detection and description of pre- and postsynaptic forms of drug-induced neurotoxicity are exemplified using recently published accounts . The inclusion of microglial markers into research strategies has widened contemporary pathogenetic concepts on methamphetamine ( MA ) -induced neurotoxicity as an example of inflammatory neurodegeneration , thus complementing the traditional ROS and RNS-dependent stress models . Amphetamine-type neurotoxicity studies may assist in elaborating of preventive strategies for human neurodegenerative disorders . DB00131 cyclase 1 ( Q08828 ) mutations cause recessive hearing impairment in humans and defects in hair cell function and hearing in zebrafish . DB02527 ( DB02527 ) production , which is important for mechanotransduction within the inner ear , is catalyzed by adenylate cyclases ( AC ) . However , knowledge of the role of ACs in hearing is limited . Previously , a novel autosomal recessive non-syndromic hearing impairment locus DFNB44 was mapped to chromosome 7p14.1-q11.22 in a consanguineous family from Pakistan . Through whole-exome sequencing of DNA samples from hearing-impaired family members , a nonsense mutation c.3112C > T ( p.Arg1038* ) within adenylate cyclase 1 ( Q08828 ) was identified . This stop-gained mutation segregated with hearing impairment within the family and was not identified in ethnically matched controls or within variant databases . This mutation is predicted to cause the loss of 82 amino acids from the carboxyl tail , including highly conserved residues within the catalytic domain , plus a calmodulin-stimulation defect , both of which are expected to decrease enzymatic efficiency . Individuals who are homozygous for this mutation had symmetric , mild-to-moderate mixed hearing impairment . Zebrafish adcy1b morphants had no FM1-43 dye uptake and lacked startle response , indicating hair cell dysfunction and gross hearing impairment . In the mouse , Adcy1 expression was observed throughout inner ear development and maturation . Q08828 was localized to the cytoplasm of supporting cells and hair cells of the cochlea and vestibule and also to cochlear hair cell nuclei and stereocilia . Ex vivo studies in COS-7 cells suggest that the carboxyl tail of Q08828 is essential for localization to actin-based microvilli . These results demonstrate that Q08828 has an evolutionarily conserved role in hearing and that DB02527 signaling is important to hair cell function within the inner ear . P01286 as an agonist of the ghrelin receptor GHS-R1a . Ghrelin synergizes with growth hormone-releasing hormone ( P01286 ) to potentiate growth hormone ( GH ) response through a mechanism not yet fully characterized . This study was conducted to analyze the role of P01286 as a potential ligand of the ghrelin receptor , GHS-R1a . The results show that hGHRH(1-29)NH(2) ( P01286 ) induces a dose-dependent calcium mobilization in P29320 293 cells stably transfected with GHS-R1a an effect not observed in wild-type P29320 293 cells . This calcium rise is also observed using the Q02643 agonists JI-34 and JI-36 . Radioligand binding and cross-linking studies revealed that calcium response to P01286 is mediated by the ghrelin receptor GHS-R1a . P01286 activates the signaling route of inositol phosphate and potentiates the maximal response to ghrelin measured in inositol phosphate turnover . The presence of P01286 increases the binding capacity of (125)I-ghrelin in a dose dependent-fashion showing a positive binding cooperativity . In addition , confocal microscopy in CHO cells transfected with GHS-R1a tagged with enhanced green fluorescent protein shows that P01286 activates the GHS-R1a endocytosis . Furthermore , the selective P01286 -R antagonists , JV-1-42 and JMR-132 , act also as antagonists of the ghrelin receptor GHS-R1a . Our findings suggest that P01286 interacts with ghrelin receptor GHS-R1a , and , in consequence , modifies the ghrelin-associated intracellular signaling pathway . This interaction may represent a form of regulation , which could play a putative role in the physiology of GH regulation and appetite control . Phosphodiesterase-4 influences the PKA phosphorylation status and membrane translocation of G-protein receptor kinase 2 ( P25098 ) in P29320 -293beta2 cells and cardiac myocytes . Membrane-recruitment of P25098 ( G-protein receptor kinase 2 ) provides a fundamental step in the desensitization process controlling GPCRs ( G-protein-coupled receptors ) , such as the beta2AR ( beta2-adrenergic receptor ) . In the present paper , we show that challenge of P29320 -293beta2 [ human embryonic kidney cells stably overexpressing the FLAG-tagged beta2AR-GFP ( green fluorescent protein ) ] cells with the beta-adrenoceptor agonist , isoprenaline , causes P25098 to become phosphorylated by PKA ( DB02527 -dependent protein kinase ) . This action is facilitated when DB02527 -specific DB05876 ( phosphodiesterase-4 ) activity is selectively inactivated , either chemically with rolipram or by siRNA ( small interfering RNA ) -mediated knockdown of Q07343 and Q08499 . DB05876 -selective inhibition by rolipram facilitates the isoprenaline-induced membrane translocation of P25098 , phosphorylation of the beta2AR by P25098 , membrane translocation of beta-arrestin and internalization of beta2ARs . DB05876 -selective inhibition also enhances the ability of isoprenaline to trigger the PKA phosphorylation of P25098 in cardiac myocytes . In the absence of isoprenaline , rolipram-induced inhibition of DB05876 activity in P29320 -293beta2 cells acts to stimulate PKA phosphorylation of P25098 , with consequential effects on P25098 membrane recruitment and P25098 -mediated phosphorylation of the beta2AR . We propose that a key role for DB05876 enzymes is : ( i ) to gate the action of PKA on P25098 , influencing the rate of P25098 phosphorylation of the beta2AR and consequential recruitment of beta-arrestin subsequent to beta-adrenoceptor agonist challenge , and ( ii ) to protect P25098 from inappropriate membrane recruitment in unstimulated cells through its phosphorylation by PKA in response to fluctuations in basal levels of DB02527 . Genomic structure and chromosome location of the murine Q01064 phosphodiesterase gene . Cyclic nucleotide phosphodiesterases ( PDEs ) catalyze the hydrolysis of DB02527 and cGMP , thereby participating in regulation of the intracellular concentrations of these second messengers . The PDE1 family is defined by regulation of activity by calcium and calmodulin . We have cloned and characterized the mouse Q01064 gene , which encodes the 63-kDa calcium/calmodulin-dependent PDE ( P62158 -PDE ) , an isozyme that is expressed in the CNS in the olfactory tract , dentate gyrus , and striatum and may participate in learning , memory , and regulation of phosphorylation of Q9UD71 in dopaminergic neurons . We screened an I-129/SvJ mouse genomic library and identified exons 2-13 of the Q01064 gene that span 8.4 kb of genomic DNA . Exons range from 67 to 205 nucleotides and introns from 91 to 2250 nucleotides in length . Exon 1 was not present in the 3 kb of genomic DNA 5' to exon 2 in our clones . The mouse Q01064 gene shares many similar or identical exon boundaries as well as considerable sequence identity with the rat Q07343 and Q08499 genes and the Drosophila dunce DB02527 -specific PDE gene dnc , suggesting that these genes all arose from a common ancestor . Using fluorescence in situ hybridization , we localized the Q01064 gene to the distal tip of mouse Chromosome ( Chr ) 15 . Effects of DB00482 and DB00744 on liver metastasis and lipidperoxidation in pancreatic cancer in Syrian hamsters . Selective inhibition of eicosanoid synthesis is thought to have effects on carcinogenesis in lung and colon cancer . However , it is still unknown whether pancreatic cancer might also be influenced . Therefore we evaluated the impact of selective cyclooxygenase-2 inhibitor DB00482 and selective P09917 inhibitor DB00744 on liver metastasis in a solid model of pancreatic adenocarcinoma in Syrian hamster . In week 33 , the animals were sacrificed and incidence of pancreatic carcinomas and number and size of liver metastases were determined . Activities of antioxidative enzymes ( GSHPX/SOD ) and concentrations of products of lipidperoxidation were measured in liver metastases and non-metastatic hepatic tissue . The incidence ( 54.5 vs. 100 % ) , number ( 3.17 +/- 0.98 vs. 6.75 +/- 0.71 ) and size ( 2.67 +/- 1.97 vs. 11.75 +/- 1.98 mm2 ) of liver metastases were decreased by combined therapy of DB00744 and DB00482 ( P < 0.05 ) . Furthermore , activities of GSHPX ( [ 73.77 +/- 5.67 ] *10(5) vs. [ 15.49 +/- 4.02 ] *10(5) U/mg prot. ; P < 0.05 ) and SOD ( 474.92 +/- 108.8 vs. 127.89 +/- 38.75 U/mg prot. ; P < 0.05 ) were increased , while lipidperoxidation ( 0.31 +/- 0.08 nmol/mg prot. vs. 1.54 +/- 0.55 nmol/mg prot. ; P < 0.05 ) was decreased by combination therapy , in non-metastatic hepatic tissue . Moreover , combined therapy increased lipidperoxidation in liver metastases ( 0.47 +/- 0.09 vs. 1.95 +/- 0.12 nmol/mg prot. ; P < 0.05 ) . Thus , a combination of DB00482 and DB00744 might be a new concept to decrease tumour growth in liver metastases in advanced pancreatic cancer . Trichinella spiralis and Trichinella pseudospiralis : developmental patterns of enzymes involved in thymidylate biosynthesis and pyrimidine salvage . P04818 , dihydrofolate reductase and P33316 specific activities were found to remain at a high and constant level in crude extracts from adult worms of Trichinella spiralis , as well as from muscle larvae of both Trichinella spiralis ( isolated 1-24 months after infection ) and Trichinella pseudospiralis ( isolated 5.5-13 months after infection ) . The results obtained with Trichinella pseudospiralis muscle larvae isolated with the use of pepsin did not differ from those obtained when pepsin was not used . No thymidine kinase activity could be detected in muscle larvae of either species and thymidine phosphorylase could be found only in T. pseudospiralis larvae isolated without the use of pepsin . Muscle larvae of both species contained orotidylate phosphoribosyl transferase activity , pointing to a possibility of 5-fluorouracil activation . DB02745 phosphorylase , another enzyme involved in 5-fluorouracil anabolism , was also present in T. pseudospiralis muscle larvae . Results of comparative studies on inhibition of purified T. spiralis and rat thymidylate synthases by substrate ( 4-thio-5-fluoro- DB03800 , 2-thio-5-fluoro-dCMP and N4-hydroxy-dCMP ) and cofactor ( ZD 9331 ) analogues indicated only DB03800 analogues to show feeble selectivity towards the parasite enzyme . A hypothesis is discussed , assuming high expression of thymidylate synthase in muscle larvae to be connected with their cells being arrested in the cell cycle . Characterization of the human adenylyl cyclase gene family : cDNA , gene structure , and tissue distribution of the nine isoforms . The membrane-bound adenylyl cyclases ( ACs ) represent one of the major families of effector enzymes for G protein-coupled receptors . Eight human AC isoforms , encoded by separate genes , have been identified up to now . However , in several cases only partial cDNA sequences are available ( Q08828 ,2,5 ) . A ninth expected isoform , the human ortholog of rat Q8NFM4 , has not been described yet . Using the high inter-species homology of mammalian AC isoforms , we searched the human genome and we succeeded to identify full-length coding sequences for all enzymes . Where required , missing sequence information was provided experimentally . Analysis of genomic sequences from the Celera database also allowed us to determine the exon-intron boundaries for Q08828 -9 and to establish the gene structures . We found that human AC genes comprise 11 to 26 exons , which are distributed over 16 to 430kb . We further report expression profiles for the nine ACs in a panel of 16 human tissues and in human embryonic kidney ( P29320 ) cells . Phosphodiesterase 4 inhibitor cilomilast inhibits fibroblast-mediated collagen gel degradation induced by tumor necrosis factor-alpha and neutrophil elastase . Tissue destruction , resulting in emphysema , can be a consequence of several pathologic processes . The current study evaluated the effects of the phosphodiesterase (PDE)4 inhibitor , cilomilast , and other PDE inhibitors on the ability of fibroblasts to degrade extracellular matrix . Using the three-dimensional collagen gel culture system , fibroblasts ( HFL-1 ) were cultured with tumor necrosis factor ( P01375 ) -alpha , known to induce matrix metalloproteinase ( MMP ) release , and/or neutrophil elastase ( NE ) , which can induce MMP activation . On Day 4 , gels containing P01375 and NE were significantly degraded ( 20.8 +/- 2.9 % of original collagen content ) . DB03849 ( 10 micro M ) inhibited this degradation ( 84.4 +/- 8.4 % ) . DB01427 , a PDE3 inhibitor , and zaprinast , a O76074 inhibitor , had no effect . Gelatin zymography and immunoblotting revealed that fibroblasts cultured with P01375 released increased amounts of latent P03956 and -9 . The addition of NE resulted in the conversion of P03956 and -9 to their active forms , indicative of collagen degradation . DB03849 inhibited the release of P03956 and -9 , as well as conversion of P03956 to its active form . Using real-time PCR analysis , cilomilast 's effect on P03956 release was not associated with the proteinase 's mRNA expression , suggesting that the inhibition of release is regulated at the post-transcriptional level . These results suggest that cilomilast may be a potentially effective therapeutic agent in diseases characterized by excessive tissue destruction , such as emphysema .