MH_dev_303

Preliminary X-ray crystallographic studies of UDP-glucose - 4 - epimerase from Aspergillus nidulans . UDP-glucose - 4 - epimerase ( Q14376 REA ) from Aspergillus nidulans was overexpressed in Escherichia coli , purified via DB00117 - tag affinity chromatography and cocrystallized with DB03501 MEN using the microbatch method . The crystals diffracted to 2.4 Å resolution using synchrotron radiation on the Canadian Light Source 08ID - 1 beamline . Examination of the data with d * O95069 REA revealed nonmerohedral twinning , from which a single lattice was ultimately extracted for processing . The final space group was found to be P06681 REA , with unit-cell parameters a = 66.13 , b = 119.15 , c = 161.42 Å , β = 98.48 ° . An initial structure solution has been obtained via molecular replacement employing human Q14376 REA ( PDB entry 1hzj ) as a template model .

Microarray analysis revealed different gene expression patterns in HepG 2 cells treated with low and high concentrations of the extracts of Anacardium occidentale shoots . In this study , the effects of low and high concentrations of the Anacardium occidentale shoot extracts on gene expression in liver HepG 2 cells were investigated . From MTT assays , the concentration of the shoot extracts that maintained 50 % cell viability ( IC ( 50 ) ) was 1.7 mg / ml . Cell viability was kept above 90 % at both 0.4 mg / ml and 0.6 mg / ml of the extracts . The three concentrations were subsequently used for the gene expression analysis using Affymetrix Human Genome 1.0 S . T arrays . The microarray data were validated using real-time qRT-PCR . A total of 246 , 696 and 4503 genes were significantly regulated ( P < 0.01 ) by at least 1.5- fold in response to 0.4 , 0.6 and 1.7 mg / ml of the extracts , respectively . Mutually regulated genes in response to the three concentrations included CDKN 3 , LOC 10028961 2 , P00374 REA , Q99986 REA , Q99741 REA , Q96GD4 and P78334 . Genes like Q07973 REA , P38398 REA , O14965 REA , P06493 REA , P24941 REA , P11802 REA and P06213 REA were significantly regulated at 0.6 mg / ml and 1.7 mg but not at 0.4 mg / ml . However , the expression of genes including O75473 REA , P17936 REA , P06400 REA , P14735 REA , P01130 REA , P55157 REA , P04114 REA , MTIX , P04179 REA and P08294 REA were exclusively regulated at the IC ( 50 ) concentration . In conclusion , low concentrations of the extracts were able to significantly regulate a sizable number of genes . The type of genes that were expressed was highly dependent on the concentration of the extracts used .

DB00203 SUB induces angiogenic response in human coronary arteriolar endothelial cells through the expression of thioredoxin , hemeoxygenase and vascular endothelial growth factor . This study was undertaken to investigate the effect of phosphodiesterase - 5 ( O76074 REA ) inhibitor , sildenafil , on angiogenic response in human coronary arteriolar endothelial cells ( HCAEC ) . The cells exposed to sildenafil ( 1-20 microM ) demonstrated significantly accelerated tubular morphogenesis with the induction of thioredoxin - 1 ( P10599 REA - 1 ) , hemeoxygenase - 1 ( P09601 REA ) and P15692 REA . DB00203 SUB induced P15692 REA and angiopoietin specific receptors such as P35968 REA , Tie - 1 and Tie - 2 . This angiogenic response was repressed by tinprotoporphyrin IX ( SnPP ) , an inhibitor of P09601 REA enzyme activity . DB00203 SUB below 1 muM has no angiogenic effect as evidenced by reduced tuborogenesis . DB00203 SUB along with SnPP inhibited both P15692 REA and Q15389 REA ( Ang - 1 ) protein expression . Therefore our results demonstrated for the first time that sildenafil is a very potent pro-angiogenic factor .

DB00203 SUB attenuates inflammation and oxidative stress in pelvic ganglia neurons after bilateral cavernosal nerve damage . Erectile dysfunction is a common complication for patients undergoing surgeries for prostate , bladder , and colorectal cancers , due to damage of the nerves associated with the major pelvic ganglia ( P29372 REA ) . Functional re-innervation of target organs depends on the capacity of the neurons to survive and switch towards a regenerative phenotype . O76074 REA inhibitors ( PDE 5i ) have been successfully used in promoting the recovery of erectile function after cavernosal nerve damage ( BCNR ) by up-regulating the expression of neurotrophic factors in P29372 REA . However , little is known about the effects of PDE 5i on markers of neuronal damage and oxidative stress after BCNR . This study aimed to investigate the changes in gene and protein expression profiles of inflammatory , anti-inflammatory cytokines and oxidative stress related-pathways in P29372 REA neurons after BCNR and subsequent treatment with sildenafil . Our results showed that BCNR in Fisher - 344 rats promoted up-regulation of cytokines ( interleukin - 1 ( IL - 1 ) β , P05231 REA , P22301 REA , transforming growth factor β 1 ( TGFβ 1 ) , and oxidative stress factors ( DB02701 adenine dinucleotide phosphate ( NADPH ) oxidase , P05164 REA ( P05164 REA ) , inducible nitric oxide synthase ( P35228 REA ) , P01375 REA receptor superfamily member 5 ( P25942 REA ) that were normalized by sildenafil treatment given in the drinking water . In summary , PDE 5i can attenuate the production of damaging factors and can up-regulate the expression of beneficial factors in the P29372 REA that may ameliorate neuropathic pain , promote neuroprotection , and favor nerve regeneration .

Hsp 27 regulates epithelial mesenchymal transition , metastasis , and circulating tumor cells in prostate cancer . Defining the mechanisms underlying metastatic progression of prostate cancer may lead to insights into how to decrease morbidity and mortality in this disease . An important determinant of metastasis is epithelial-to-mesenchymal transition ( EMT ) , and the mechanisms that control the process of EMT in cancer cells are still emerging . Here , we report that the molecular chaperone Hsp 27 ( P04792 REA ) drives EMT in prostate cancer , whereas its attenuation reverses EMT and decreases cell migration , invasion , and matrix metalloproteinase activity . Mechanistically , silencing Hsp 27 decreased P05231 REA - dependent P40763 REA phosphorylation , nuclear translocation , and P40763 REA binding to the Twist promoter , suggesting that Hsp 27 is required for P05231 REA - mediated EMT via modulation of P40763 REA / Twist signaling . We observed a correlation between Hsp 27 and Twist in patients with prostate cancer , with Hsp 27 and Twist expression each elevated in high-grade prostate cancer tumors . Hsp 27 inhibition by DB06094 MEN , an antisense therapy currently in phase II trials , reduced tumor metastasis in a murine model of prostate cancer . More importantly , DB06094 MEN treatment decreased the number of circulating tumor cells in patients with metastatic castration-resistant prostate cancer in a phase I clinical trial . Overall , this study defines Hsp 27 as a critical regulator of P05231 REA - dependent and P05231 REA - independent EMT , validating this chaperone as a therapeutic target to treat metastatic prostate cancer .

The P05230 REA - specific single-chain antibody scFv 1C9 effectively inhibits breast cancer tumour growth and metastasis . Immunotherapy mediated by recombinant antibodies is an effective therapeutic strategy for a variety of cancers . In a previous study , we demonstrated that the fibroblast growth factor 1 ( P05230 REA ) - specific recombinant antibody scFv 1C9 arrests the cell cycle at the G0 / P55008 transition by blocking the intracrine P05230 REA pathway in breast cancer cells . Here , we further show that the overexpression of scFv 1C9 in MCF - 7 and MDA-MB - 231 breast cancer cells by lentiviral infection resulted in decreased tumourigenicity , tumour growth and lung metastasis through P05230 REA neutralization . We found that scFv 1C9 resulted in the up-regulation of P38936 REA , which in turn inhibited the expression of P24941 REA and blocked cell cycle progression . To explore the potential role of scFv 1C9 in vivo , we delivered the gene into solid tumours by electroporation , which resulted in significant inhibition of tumour growth . In tumour tissue sections , immunohistochemical staining of the cellular proliferation marker Ki - 67 and the microvessel marker CD31 showed a reduction in the proliferative index and microvessel density , respectively , upon expression of scFv 1C9 compared with the appropriate controls . Thus , our data indicate a central role for scFv 1C9 in blocking the intracrine pathway of P05230 REA , therefore , scFv 1C9 could be developed in an effective therapeutic for breast cancer .

Phosphodiesterase - 5 inhibitor sildenafil prevents neuroinflammation , lowers beta-amyloid levels and improves cognitive performance in P05067 REA / P49768 REA transgenic mice . Memory deficit is a marker of Alzheimer ' s disease ( AD ) that has been highly associated with the dysfunction of cyclic GMP ( cGMP ) signaling and an ongoing inflammatory process . Phosphodiesterase - 5 ( O76074 REA ) inhibitors prevent the breakdown of cGMP and are currently studied as a possible target for cognitive enhancement . However , it is still unknown whether inhibition of O76074 REA reversed β-amyloid peptide ( Aβ ) - induced neuroinflammation in P05067 REA / P49768 REA transgenic ( Tg P05067 REA / P49768 REA ) mice . The present study evaluated the cognitive behaviors , inflammatory mediators , and cGMP / PKG / pCREB signaling in 15 - month-old Tg P05067 REA / P49768 REA mice and age-matched wild-type ( WT ) mice that were treated with O76074 REA inhibitor sildenafil and the inhibitor of cGMP-dependent protein kinase Rp - 8 - Br-PET-cGMPS . In comparison with WT mice , Tg P05067 REA / P49768 REA mice were characterized by impaired cognitive ability , neuroinflammatory response , and down-regulated cGMP signaling . DB00203 SUB reversed these memory deficits and cGMP / PKG / pCREB signaling dysfunction ; it also reduced both the soluble Aβ1 - 40 and Aβ1 - 42 levels in the hippocampus . These effects of sildenafil were prevented by intra-hippocampal infusion of the Rp - 8 - Br-PET-cGMPS . These results suggest that sildenafil could restore cognitive deficits in Tg P05067 REA / P49768 REA mice by the regulation of PKG / pCREB signaling , anti-inflammatory response and reduction of Aβ levels .

OSU - 03012 and Viagra Treatment Inhibits the Activity of Multiple Chaperone Proteins and Disrupts the Blood-Brain Barrier : Implications for Anti-Cancer Therapies . We examined the interaction between OSU - 03012 ( also called AR - 12 ) with phosphodiesterase 5 ( O76074 REA ) inhibitors to determine the role of the chaperone glucose-regulated protein ( P11021 REA ) / P11021 REA / P11021 REA in the cellular response . DB00203 SUB ( Viagra ) interacted in a greater than additive fashion with OSU - 03012 to kill stem-like GBM cells . Treatment of cells with OSU - 03012 / sildenafil : abolished the expression of multiple oncogenic growth factor receptors and plasma membrane drug efflux pumps and caused a rapid degradation of P11021 REA and other HSP 70 and HSP 90 family chaperone proteins . Decreased expression of plasma membrane receptors and drug efflux pumps was dependent upon enhanced Q9NZJ5 - eIF 2α - P18848 REA - P35638 REA signaling and was blocked by P11021 REA over-expression . In vivo OSU - 03012 / sildenafil was more efficacious than treatment with celecoxib and sildenafil at killing tumor cells without damaging normal tissues and in parallel reduced expression of P08183 REA and Q9UNQ0 in the normal brain . The combination of OSU - 03012 / sildenafil synergized with low concentrations of sorafenib to kill tumor cells , and with lapatinib to kill P00533 REA over-expressing tumor cells . In multiplex assays on plasma and human tumor tissue from an OSU - 03012 / sildenafil treated mouse , we noted a profound reduction in uPA signaling and identified FGF and P23458 REA / 2 as response biomarkers for potentially suppressing the killing response . Inhibition of FGFR signaling and to a lesser extent P23458 REA / 2 signaling profoundly enhanced OSU - 03012 / sildenafil lethality .

Sulfasalazine inhibits the growth of primary brain tumors independent of nuclear factor-kappaB . Nuclear factor-kappaB ( NF-kappaB ) is a pleiotropic transcription factor that generally enhances cellular resistance to apoptotic cell death . It has been shown to be constitutively active in some cancers and is being pursued as potential anticancer target . Sulfasalazine which is used clinically to treat Crohn ' s disease has emerged as a potential inhibitor of NF-kappaB and has shown promising results in two pre-clinical studies to target primary brain tumors , gliomas . Once digested , sulfasalazine is cleaved into sulfapyridine and DB00244 MEN ( DB00244 MEN ; mesalamine ) by colonic bacteria , and the latter , too , is reported to suppress NF-kappaB activity . We now show that glioma cells obtained from patient biopsies or glioma cell lines do not show significant constitutive NF-kappaB activation , unless exposed to inflammatory cytokines . This does not change when gliomas are implanted into the cerebrum of severe combined immun-deficient mice . Nevertheless , sulfasalazine but not its cleaved form DB00244 MEN caused a dose-dependent inhibition of glioma growth . This effect was entirely attributable to the inhibition of cystine uptake via the system x ( c ) ( - ) cystine-glutamate transporter . It could be mimicked by S - 4 - carboxy-phenylglycine ( S - 4 - CPG ) a more specific system x ( c ) ( - ) inhibitor , and lentiviral expression of a constitutively active form of O15111 REA b was unable to overcome the growth retarding effects of sulfasalazine or S - 4 - CPG . Both drugs inhibited cystine uptake causing a chronic depletion of intracellular DB00143 and consequently compromised cellular redox defense which stymied tumor growth . This data suggests that system x ( c ) ( - ) is a promising therapeutic target in gliomas and possibly other cancers and that it can be pharmacologically inhibited by Sulfasalazine , an FDA-approved drug .

Activity , pharmacological inhibition and biological regulation of 3 - hydroxy - 3 - methylglutaryl coenzyme A reductase in Trypanosoma brucei . Activity of hydroxymethylglutaryl-coenzyme A ( HMG - DB01992 ) reductase , the key enzyme in the biosynthesis of steroids and polyisoprenoids in mammalian cells , has been detected in both the bloodstream form and the culture-adapted procyclic form of Trypanosoma brucei ( 3.7 + / - 0.6 and 12.7 + / - 1.8 pmol mevalonate produced min - 1 ( mg cell protein ) - 1 , respectively ) . The enzyme activity is enriched 6 - fold in microsomal fractions . Several competitive inhibitors of mammalian P04035 REA , including synvinolin ( simvastatin ) , inhibit the multiplication of both forms of trypanosome in vitro ( IC50 , approx . 25-50 microM after 2-3 days ) . This growth inhibition is potentiated by agents interfering with the exogenous supply of cholesterol , such as antibodies blocking the low-density lipoprotein ( LDL ) receptor , or 5 microM chloroquine . Conversely , growth inhibition by synvinolin can be largely reverted either by 300 nM LDL or by products of the mevalonate pathway , such as 20 mM mevalonate and in procyclics by 100 microM squalene or cholesterol . In procyclics , low concentrations of synvinolin selectively inhibit the incorporation of [ 14C ] acetate into sterols , but not into fatty acids . These results argue for a critical role in trypanosomes of a mevalonate pathway , that is involved in the biosynthesis of sterol and probably of other metabolites . The P04035 REA activity is decreased 2 - fold in procyclics incubated with 4 mM mevalonate and increased 2 - fold in the presence of 2.5 microM synvinolin . DB00641 MEN also upregulates LDL binding up to 4 - fold . These data suggest that P04035 REA and P01130 REA expression are regulated in T . brucei as in mammalian cells , to ensure sterol homeostasis .

Alzheimer ' s disease and modern lifestyle : what is the role of stress ? This Editorial highlights a study by Baglietto-Vargas et al . 2015 published in this issue of J . Neurochem . Stress is one of the environmental factors that can contribute to Alzheimer ' s disease pathogenesis . However , the role of modern-life stress has not been investigated yet . The authors reveal that modern-life stress reduces the number of dendritic spines in the hippocampus of Alzheimer ' s disease transgenic mice . The mechanism underlying such effect involves an increase in corticotropin-releasing hormone ( P06850 REA ) release that stimulates the amyloid precursor protein ( P05067 REA ) processing and fosters the generation of Amyloid-β , which negatively affects dendritic spines .

Dimerization effect of sucrose octasulfate on rat P05230 REA . Fibroblast growth factors ( FGFs ) constitute a family of at least 23 structurally related heparin-binding proteins that are involved in regulation of cell growth , survival , differentiation and migration . DB01901 MEN ( SOS ) , a chemical analogue of heparin , has been demonstrated to activate FGF signalling pathways . The structure of rat P05230 REA crystallized in the presence of SOS has been determined at 2.2 A resolution . SOS-mediated dimerization of P05230 REA was observed , which was further supported by gel-filtration experiments . The major contributors to the sulfate-binding sites in rat P05230 REA are Lys 113 , Lys 118 , Arg 122 and Lys 128 . An arginine at position 116 is a consensus residue in mammalian FGF molecules ; however , it is a serine in rat P05230 REA . This difference may be important for SOS-mediated P05230 REA dimerization in rat .

Enhancement of auranofin-induced apoptosis in MCF - 7 human breast cells by selenocystine , a synergistic inhibitor of thioredoxin reductase . P10599 REA system plays an important role in regulation of intracellular redox balance and various signaling pathways . P30044 REA ( TrxR ) is overexpressed in many cancer cells and has been identified as a potential target of anticancer drugs . DB00995 MEN ( AF ) is potent TrxR inhibitor with novel in vitro and in vivo anticancer activities . Selenocystine ( SeC ) is a nutritionally available selenoamino acid with selective anticancer effects through induction of apoptosis . In the present study , we demonstrated the synergistic effects and the underlying molecular mechanisms of SeC in combination with AF on MCF - 7 human breast cancer cells . The results showed that SeC and AF synergistically inhibited the cancer cell growth through induction of ROS-dependent apoptosis with the involvement of mitochondrial dysfunction . DNA damage-mediated p53 phosphorylation and down-regulation of phosphorylated AKT and P29323 REA also contributed to cell apoptosis . Moreover , we demonstrated the important role of TrxR activity in the synergistic action of SeC and AF . Taken together , our results suggest the strategy to use SeC and AF in combination could be a highly efficient way to achieve anticancer synergism by targeting TrxR .

Roxithromycin is an inhibitor of human coronary artery smooth muscle cells proliferation : a potential ability to prevent coronary heart disease . Roxithromycin ( RXM ) , a macrolide antibiotic , is used in clinical trials to address secondary prevention of coronary heart disease . However , the effects of RXM on human coronary artery smooth muscle cells ( CASMC ) proliferation remain unclear . Human CASMC were stimulated with growth medium containing 5 % fetal bovine serum and growth factors . RXM at 1 or 10 microg / ml , which are relevant to the therapeutic plasma levels , significantly suppressed mitogen-induced CASMC proliferation , assessed by WST - 1 assay and cell counting . Flow cytometry analysis demonstrated that RXM suppressed mitogen-induced P55008 to S progression on cell cycle . Western blot showed that RXM inhibited phosphorylation of retinoblastoma gene products , reduced protein levels of cyclin D1 and A , and restored downregulation of cyclin-dependent kinase ( CDK ) inhibitor p27kip1 . The activities of P11802 REA and P24941 REA were suppressed by RXM without affecting their protein levels . When transfected with both O15111 REA alpha and beta constructs as nuclear factor-kappa B ( NF-kappaB ) activator , CASMC entered S phase at 24 h , and RXM inhibited it . Electrophoretic mobility shift assay and immunostaining of NF-kappaB p65 demonstrated that RXM inhibited mitogen-induced NF-kappaB activation . These results indicate that RXM is an inhibitor of human CASMC proliferation through modulating cell cycle regulatory proteins and inhibiting NF-kappaB signaling pathway .

Disrupted coordinate regulation of farnesoid X receptor target genes in a patient with cerebrotendinous xanthomatosis . Cerebrotendinous xanthomatosis ( CTX ) , sterol 27 - hydroxylase ( Q02318 REA ) deficiency , is associated with markedly reduced chenodeoxycholic acid ( DB06777 MEN ) , the most powerful activating ligand for farnesoid X receptor ( Q96RI1 ) . We investigated the effects of reduced DB06777 MEN on Q96RI1 target genes in humans . Liver specimens from an untreated CTX patient and 10 control subjects were studied . In the patient , hepatic DB06777 MEN concentration was markedly reduced but the bile alcohol level exceeded DB06777 MEN levels in control subjects ( 73.5 vs . 37.8 + / - 6.2 nmol / g liver ) . DB04540 7alpha - hydroxylase ( P22680 REA ) and Na + / taurocholate-cotransporting polypeptide ( Q14973 REA ) were upregulated 84 - and 8 - fold , respectively . However , small heterodimer partner ( Q15466 REA ) and bile salt export pump were normally expressed . Marked P22680 REA induction with normal Q15466 REA expression was not explained by the regulation of liver X receptor alpha ( LXRalpha ) or pregnane X receptor . However , another nuclear receptor , hepatocyte nuclear factor 4alpha ( HNF 4alpha ) , was induced 2.9- fold in CTX , which was associated with enhanced mRNA levels of HNF 4alpha target genes , P22680 REA , 7alpha - hydroxy - 4 - cholesten - 3 - one 12alpha - hydroxylase , Q02318 REA , and Q14973 REA . In conclusion , the coordinate regulation of Q96RI1 target genes was lost in CTX . The mechanism of the disruption may be explained by a normally stimulated Q96RI1 pathway attributable to markedly increased bile alcohols with activation of HNF 4alpha caused by reduced bile acids in CTX liver .

Inhibition of neuronal nitric oxide reduces anxiety-like responses to pair housing . Many psychological disorders are characterized by anxiety and alterations in social interactions . Recent studies demonstrate that the chemical messenger nitric oxide ( NO ) can regulate both anxiety and social behaviours . We tested whether an enzyme that produces NO in the brain , neuronal nitric oxide synthase ( P29475 REA ) , serves as an interface between social interactions and anxiety-like behaviour . Several investigators have observed that mice increase anxiety-like responses in the elevated plus-maze after pair housing . P29475 REA gene deletion and DB01997 MEN were used to inhibit the production of neuronal NO . Similar to previous studies , pair housing reduced open arm exploration in the elevated plus-maze . Pair housing also increased corticotropin-releasing hormone ( P06850 REA ) immunoreactive cells in the paraventricular nucleus ( PVN ) of the hypothalamus . Inhibition of NO production increased open arm exploration in pair-housed mice but decreased open arm exploration in individually housed mice . These results suggest that the effect of P29475 REA inhibition on anxiety-like responses is context dependent and that behavioural responses to social housing are altered after P29475 REA inhibition . This research suggests that NO may play an important role in mediating the effect social interactions have on anxiety .

P30532 REA gene D398N polymorphism in Japanese lung adenocarcinoma . BACKGROUND : Recently , to identify genetic factors that modify lung cancer risk , P30532 REA non-synonymous variant amino acid position 398 ( D398N ) was identified . The site was a highly conserved in the second cellular loop of the nicotinic acetylcholine receptor subunit protein . MATERIALS AND METHODS : We have investigated P30532 REA gene polymorphism status in 302 surgically treated lung adenocarcinoma cases from Nagoya City University Hospital . The presence or absence of P30532 REA polymorphism was analyzed by direct sequences . P00533 REA mutations status was already investigated and reported . RESULTS : We detected nine cases ( 2.98 % ) of P30532 REA polymorphism ( D398N ) in our cohort . Total P00533 REA mutations were present in 129 patients ( 42.7 % ) . The polymorphism statuses were not correlated with gender ( women ; 2.1 % versus men ; 3.7 % , P = 0.5119 ) , smoking status ( never smoker ; 2.0 % versus smoker ; 4.0 % , P = 0.3339 ) , pathological stages ( stage I ; 2.6 % versus stage II-IV ; 3.8 % , P = 0.7246 ) , and P00533 REA mutation status of the lung adenocarcinomas ( mutation ; 2.3 % versus wild type ; 3.7 % , P = 0.7373 ) . In this analysis , P30532 REA polymorphism ( D398N ) patients had significantly worse prognosis ( 5/9 were dead ; mean survival = 27.1 mo ) than the patients with P30532 REA wild type ( 74/293 were dead ; mean survival = 113.9 mo ) ( log-rank test ; P = 0.0146 ) . CONCLUSION : Although P30532 REA polymorphism is rare from Japanese lung cancer , polymorphism status might be correlated with shorter survival .

Partial agonists of the α3β4 * neuronal nicotinic acetylcholine receptor reduce ethanol consumption and seeking in rats . DB00898 MENMAX DB00898 MEN use disorders ( AUDs ) impact millions of individuals and there remain few effective treatment strategies . Despite evidence that neuronal nicotinic acetylcholine receptors ( nAChRs ) have a role in AUDs , it has not been established which subtypes of the nAChR are involved . Recent human genetic association studies have implicated the gene cluster P32297 REA - P30532 REA - P30926 REA encoding the α3 , α5 , and β4 subunits of the nAChR in susceptibility to develop nicotine and alcohol dependence ; however , their role in ethanol-mediated behaviors is unknown due to the lack of suitable and selective research tools . To determine the role of the α3 , and β4 subunits of the nAChR in ethanol self-administration , we developed and characterized high-affinity partial agonists at α3β4 nAChRs , CP - 601932 , and PF - 4575180 . Both CP - 601932 and PF - 4575180 selectively decrease ethanol but not sucrose consumption and operant self-administration following long-term exposure . We show that the functional potencies of CP - 601932 and PF - 4575180 at α3β4 nAChRs correlate with their unbound rat brain concentrations , suggesting that the effects on ethanol self-administration are mediated via interaction with α3β4 nAChRs . Also varenicline , an approved smoking cessation aid previously shown to decrease ethanol consumption and seeking in rats and mice , reduces ethanol intake at unbound brain concentrations that allow functional interactions with α3β4 nAChRs . Furthermore , the selective α4β2 ( * ) nAChR antagonist , DHβE , did not reduce ethanol intake . Together , these data provide further support for the human genetic association studies , implicating P32297 REA and P30926 REA genes in ethanol-mediated behaviors . CP - 601932 has been shown to be safe in humans and may represent a potential novel treatment for AUDs .

Differential expression of cholesterol hydroxylases in Alzheimer ' s disease . DB04540 is eliminated from neurons by oxidization , which generates oxysterols . DB04540 oxidation is mediated by the enzymes cholesterol 24 - hydroxylase ( Q9Y6A2 REA ) and cholesterol 27 - hydroxylase ( Q02318 REA ) . Immunocytochemical studies show that Q9Y6A2 REA and Q02318 REA are expressed in neurons and some astrocytes in the normal brain , and Q02318 REA is present in oligodendrocytes . In Alzheimer ' s disease ( AD ) , Q9Y6A2 REA shows prominent expression in astrocytes and around amyloid plaques , whereas Q02318 REA expression decreases in neurons and is not apparent around amyloid plaques but increases in oligodendrocytes . Although previous studies have examined the effects of synthetic oxysterols on the processing of amyloid precursor protein ( P05067 REA ) , the actions of the naturally occurring oxysterols have yet to be examined . To understand the role of cholesterol oxidation in AD , we compared the effects of 24 ( S ) - and 27 - hydroxycholesterol on the processing of P05067 REA and analyzed the cell-specific expression patterns of the two cholesterol hydroxylases in the human brain . Both oxysterols inhibited production of Abeta in neurons , but 24 ( S ) - hydroxycholesterol was approximately 1000 - fold more potent than 27 - hydroxycholesterol . The IC ( 50 ) of 24 ( S ) - hydroxycholesterol for inhibiting Abeta secretion was approximately 1 nm . Both oxysterols induced O95477 REA expression with IC ( 50 ) values similar to that for inhibition of A beta secretion , suggesting the involvement of liver X receptor . Oxysterols also inhibited protein kinase C activity and P05067 REA secretion following stimulation of protein kinase C . The selective expression of Q9Y6A2 REA around neuritic plaques and the potent inhibition of P05067 REA processing in neurons by 24 ( S ) - hydroxycholesterol suggests that Q9Y6A2 REA affects the pathophysiology of AD and provides insight into how polymorphisms in the Q9Y6A2 REA gene might influence the pathophysiology of this prevalent disease .

DB00203 SUB inhibits calcineurin / Q13469 REA - 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 REA ) 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 REA expressions were determined by Western blot . P24941 REA ( P24941 REA ) 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 REA 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 REA protein by siRNA transfection abolished serotonin-induced cyclin A expression and consequent P24941 REA activation and DNA synthesis . We further found that pretreatment of cells with sildenafil suppressed serotonin-triggered activation of calcineurin / Q13469 REA signaling pathway and resultant cyclin A expression , P24941 REA 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 REA cascade-mediated cyclin A expression , P24941 REA activation and PASMC proliferation to contribute to the overall effects of sildenafil in the treatment of pulmonary hypertension .

Significance of water molecules in the inhibition of cylin-dependent kinase 2 and 5 complexes . Interest in P24941 REA and Q00535 REA has stemmed mainly from their association with cancer and neuronal migration or differentiation related diseases and the need to design selective inhibitors for these kinases . In the present paper , eight Molecular Dynamics ( MD ) simulations are carried out to examine the importance of structure and dynamics of water in the active site of both P24941 REA and Q00535 REA complexes with roscovitine and indirubin analogues . Together with previous results , the current work shows a highly conserved water-involved hydrogen bonding ( HB ) network in both P24941 REA - and Q00535 REA - indirubin combinations to complete information from the X-ray crystallography . The simulations suggest the importance of such a network for combining the inhibitor to the host protein as well as the significance of using an activated CDK as a template when designing new inhibitors . Different binding patterns of roscovitine in P24941 REA and Q00535 REA are detected during the simulations because of the different binding conformations of the group on the P06681 REA side chain , which might offer a clue toward finding highly selective inhibitors with regards to P24941 REA and Q00535 REA .