MH_dev_291

The phosphatidylinositol 3 - kinase / protein kinase B signaling pathway is activated by lipoteichoic acid and plays a role in Kupffer cell production of interleukin - 6 ( P05231 REA ) and P22301 REA . Sepsis caused by gram-positive bacteria lacking lipopolysaccharide ( LPS ) has become a major and increasing cause of mortality in intensive-care units . We have recently demonstrated that the gram-positive-specific bacterial cell wall component lipoteichoic acid ( P01374 REA ) stimulates the release of the proinflammatory cytokines in Kupffer cells in culture . In the present study , we have started to assess the signal transduction events by which P01374 REA induces the production of tumor necrosis factor alpha ( P01375 REA ) , interleukin - 6 ( P05231 REA ) , and the anti-inflammatory cytokine P22301 REA in rat Kupffer cells . P01374 REA was found to trigger phosphorylation of mitogen-activated protein kinases ( MAPK ) ( p38 MAPK and P29323 REA 1/2 ) and protein kinase B ( P31749 REA ) . Compared to LPS , P01374 REA was more potent in inducing P31749 REA phosphorylation after 40 min , although we found that the cytokine responses were similar . For both bacterial molecules , blocking phosphatidylinositol 3 - kinase ( P19957 REA - K ; Ly294002 ) or O60674 REA ( O60674 REA ; AG490 ) particularly affected the induction of P05231 REA and P22301 REA release , whereas P01375 REA levels were strongly reduced by inhibition of Src family tyrosine kinases ( Q99463 ) . All three cytokines were reduced by inhibition of p38 MAPK ( SB202190 ) or the broad-range tyrosine kinase inhibitor genistein , whereas P05231 REA release was particularly blocked by inhibition of P29323 REA 1/2 ( PD98059 ) . Divergences in the regulatory pathways controlling P01375 REA , P22301 REA , and P05231 REA production in Kupffer cells following LPS or P01374 REA stimulation may create a basis for understanding how the balance between pro - and anti-inflammatory cytokines is regulated in the liver following infections by gram-positive or gram-negative bacteria .

Serotonin skews human macrophage polarization through P41595 REA and P34969 REA . Besides its role as a neurotransmitter , serotonin ( 5 - hydroxytryptamine , 5HT ) regulates inflammation and tissue repair via a set of receptors ( 5HT ( 1-7 ) ) whose pattern of expression varies among cell lineages . Considering the importance of macrophage polarization plasticity for inflammatory responses and tissue repair , we evaluated whether 5HT modulates human macrophage polarization . 5HT inhibited the LPS-induced release of proinflammatory cytokines without affecting P22301 REA production , upregulated the expression of M2 polarization-associated genes ( P05120 REA , P07996 REA , Q9NY15 , Q86Y22 REA ) , and reduced the expression of M1 - associated genes ( P08476 REA , P41597 REA , P39900 REA , P05121 REA , P29016 REA , O94788 REA ) . Whereas only 5HT ( 7 ) mediated the inhibitory action of 5HT on the release of proinflammatory cytokines , both 5HT ( 2B ) and 5HT ( 7 ) receptors mediated the pro-M 2 skewing effect of 5HT . In fact , blockade of both receptors during in vitro monocyte-to-macrophage differentiation preferentially modulated the acquisition of M2 polarization markers . 5HT ( 2B ) was found to be preferentially expressed by anti-inflammatory M2 ( P09603 REA ) macrophages and was detected in vivo in liver Kupffer cells and in tumor-associated macrophages . Therefore , 5HT modulates macrophage polarization and contributes to the maintenance of an anti-inflammatory state via 5HT ( 2B ) and 5HT ( 7 ) , whose identification as functionally relevant markers for anti-inflammatory / homeostatic human M2 macrophages suggests their potential therapeutic value in inflammatory pathologies .

Dopamine agonist-induced hypothermia and disruption of prepulse inhibition : evidence for a role of D3 receptors ? The dopamine D3 / D2 receptor agonists 7 - OH-DPAT , quinpirole , quinelorane , and PD128907 , the mixed dopamine agonist apomorphine , the D2 agonist bromocriptine , and the D1 / D5 agonist SKF 38393 were examined in models of hypothermia and prepulse inhibition ( PPI ) in Wistar rats . As dopamine agonist-induced hypothermia has been proposed as a model of D3 receptor function , and dopamine agonists are known to disrupt PPI , drug potencies to induce hypothermia were established and compared with doses necessary to disrupt PPI . 7 - OH-DPAT , quinpirole , quinelorane , PD128907 , and apomorphine , reduced body temperature and disrupted PPI with a similar rank order of potency ( quinelorane > quinpirole = 7 - OH-DPAT > PD128907 = apomorphine ) . DB01200 SUB and SKF 38393 were ineffective in both models . In a separate study , the dopamine reuptake inhibitors cocaine and GBR 12909 had no effect on PPI . In a final set of studies , the D2 / D3 antagonist raclopride blocked both 7 - OH-DPAT-induced hypothermia and 7 - OH-DPAT-induced PPI disruption . The P08908 REA antagonist WAY 100,135 , and the peripheral D2 - like antagonist domperidone had no effect . These findings suggest that the hypothermia and PPI disruptions seen with some of these dopamine agonists may be mediated by central D3 receptors ; however , only studies using more selective dopamine receptor ligands can definitively rule out effects at the D2 or D4 receptors .

Neurological impairment in experimental antiphospholipid syndrome is associated with increased ligand binding to hippocampal and cortical serotonergic P08908 REA receptors . The antiphospholipid syndrome ( APS ) is an autoimmune disease where the presence of high titers of circulating autoantibodies causes thrombosis with consecutive infarcts . In experimental APS ( eAPS ) , a mouse model of APS , behavioral abnormalities develop in the absence of vessel occlusion or infarcts . Using brain hemispheres of control and eAPS mice with documented neurological and cognitive deficits , we checked for lymphocytic infiltration , activation of glia and macrophages , as well as alterations of ligand binding densities of various neurotransmitter receptors to unravel the molecular basis of this abnormal behavior . Lymphocytic infiltrates were immunohistochemically characterized using antibodies against CD3 , P01730 REA , CD8 and forkhead box P09131 ( Foxp 3 ) , respectively . P14136 REA , Iba 1 and P34810 REA - immunohistochemistry was performed , to check for activation of astrocytes , microglia and macrophages . Ligand binding densities of DB01221 , AMPA , GABAA and P08908 REA receptors were analyzed by in vitro receptor autoradiography . No significant inflammatory reaction occurred in eAPS mice . There was neither activation of astrocytes or microglia nor accumulation of macrophages . Binding values of excitatory and inhibitory neurotransmitter receptors were largely unchanged . However , ligand binding densities of the modulatory serotonergic P08908 REA receptors in the hippocampus and in the primary somatosensory cortex of eAPS mice were significantly upregulated which is suggested to induce the behavioral abnormalities observed .

P06401 REA modulator DB05366 MEN induces extracellular matrix metalloproteinase inducer in cultured human uterine leiomyoma cells . Effects of progesterone receptor modulator DB05366 MEN on the expression of the extracellular matrix ( Q13201 REA ) components were examined in cultured human uterine leiomyoma and myometrial cells . Q13201 REA metalloproteinase inducer ( P35613 REA ) , matrix metalloproteinases ( MMPs ) , tissue inhibitors of MMP ( TIMPs ) and collagen levels were assessed by Western blot analysis , MMP activity assay and real-time RT-PCR . RNA interference ( RNAi ) of P35613 REA was performed using small interfering mRNA . In cultured leiomyoma cells , DB05366 MEN treatment at concentrations greater than or equal to 10 (-8 ) M significantly increased P35613 REA , P03956 REA and P22894 REA protein contents and P03956 REA , P08253 REA , P08254 REA and P14780 REA mRNA levels , and activity of P03956 REA , P08253 REA , P08254 REA and P14780 REA in the medium . P01033 REA and P16035 REA were significantly decreased at mRNA and protein levels by DB05366 MEN treatment at concentrations > or = 10 ( - 7 ) M in these cells . DB05366 MEN treatment decreased types I and III collagen protein contents . However , DB05366 MEN treatment did not affect the Q13201 REA component expression in cultured myometrial cells . RNAi of P35613 REA abrogated DB05366 MEN - mediated both induction of MMPs and reduction of TIMPs and collagens in cultured leiomyoma cells . These results suggest that DB05366 MEN modulates the expression of P35613 REA , MMPs , TIMPs and collagens in cultured leiomyoma cells without comparable effects on myometrial cells .

DB01411 MEN inhibits NF-kappaB activation and Q02817 REA gene expression in cultured human epithelial cells . DB01411 MEN is a selective cysteinyl leukotriene ( 1 ) ( cysLT ( 1 ) ) receptor antagonist , and is now widely used in the treatment of asthma . The anti-asthmatic effect of pranlukast may be rendered not only by antileukotriene activity , but also by other pharmacological activity . This study was designed to investigate whether pranlukast had inhibitory effects on nuclear factor-kappaB ( NF-kappaB ) activation and mucin gene expression in cultured human epithelial cells . Luciferase assay was mainly used for analysis . Cultured epithelial cells were transfected with NF-kappaB luciferase vector , Q02817 REA or P98088 REA luciferase vectors . Lipopolysaccharide ( LPS ) significantly increased NF-kappaB activation in NCI-H 292 cells , which was inhibited by the pretreatment by pranlukast in a dose-dependent manner . Either LTD ( 4 ) or pranlukast alone did not increase NF-kappaB activation in NCI-H 292 cells . DB01411 MEN also inhibited NF-kappaB activation induced by phorbol 12 - myristate 13 - acetate ( PMA ) . DB01411 MEN also significantly inhibited LPS-induced Q02817 REA mRNA expression by reverse transcription-polymerase chain reaction ( RT-PCR ) analysis in NCI-H 292 cells . DB01411 MEN also inhibited LPS-induced Q02817 REA gene expression in HM3 - Q02817 REA cells . However , pranlukast did not inhibit P98088 REA gene transcription activity induced by lipoteichoic acid ( P01374 REA ) in NCI-H 292 cells . These results suggest that pranlukast may inhibit NF-kappaB activation and Q02817 REA gene transcription through pathways distinct from cysLT ( 1 ) receptor antagonism in cultured human epithelial cells .

Control of phenylalanine and tyrosine metabolism by phosphorylation mechanisms . A system for the parallel determination of enzyme phosphorylation and expressed activity in rat liver cells , and its application to studies of phenylalanine hydroxylase and tyrosine aminotransferase , is described . DB00120 MEN hydroxylase is phosphorylated by agents which stimulate cyclic AMP - and Ca2 + - dependent protein kinase activity . The phosphorylation site ( s ) appear to be the same for both kinases . Phosphorylation is accompanied by increased metabolic flux at low , physiologically relevant , substrate concentrations . P01308 REA and spermine both inhibit the phosphorylation of the enzyme , possibly by increasing dephosphorylation . P17735 REA is phosphorylated in liver cell incubations but the rate is slow and insensitive to additions to the medium . No parallel changes in flux could be detected . Both enzymes are subject to complex regulatory mechanisms , short - and long-term . Their activities may be coordinated in vivo by control exerted at the level of the plasma membrane where both amino acids share the same transport processes . Determination of the control coefficients for the several components indicates that membrane transport may be a major limitation on flux .

Prospection of genomic regions divergently selected in racing line of Quarter Horses in relation to cutting line . Selection of Quarter Horses for different purposes has led to the formation of lines , including racing and cutting horses . The objective of this study was to identify genomic regions divergently selected in racing line of Quarter Horses in relation to cutting line applying relative extended haplotype homozygosity ( REHH ) analysis , an extension of extended haplotype homozygosity ( EHH ) analysis , and the fixation index ( F ST ) statistic . A total of 188 horses of both sexes , born between 1985 and 2009 and registered at the Brazilian Association of Quarter Horse Breeders , including 120 of the racing line and 68 of the cutting line , were genotyped using single nucleotide polymorphism arrays . On the basis of 27 genomic regions identified as selection signatures by REHH and F ST statistics , functional annotations of genes were made in order to identify those that could have been important during formation of the racing line and that could be used subsequently for the development of selection tools . Genes involved in muscle growth ( n = 8 ) , skeletal growth ( n = 10 ) , muscle energy metabolism ( n = 15 ) , cardiovascular system ( n = 14 ) and nervous system ( n = 23 ) were identified , including the O75072 , P06213 REA , P13807 REA , P35523 REA , Q15746 REA , P43405 REA , P03950 , P26992 REA and P41595 REA .

The presence and function of dopamine type 2 receptors in boar sperm : a possible role for dopamine in viability , capacitation , and modulation of sperm motility . Several studies have shown that dopamine and other catecholamines are present in oviduct luminal fluid . We recently reported that dopamine type 2 receptors ( P14416 REA ) are present in a wide range of mammalian sperm , suggesting a role for dopaminergic signaling in events such as fertilization , capacitation , and sperm motility . In the present study , we used Western blot analysis to show that boar sperm express P14416 REA and that their activation with dopamine ( 100 nM ) has a positive effect on cell viability that can be correlated with AKT / P31749 REA phosphorylation . DB01200 SUB ( 100 nM ) and dopamine ( 100 nM and 10 muM ) increased tyrosine phosphorylation during the capacitation period . Immunofluorescence analysis indicated that P14416 REA localization is dynamic and depends on the capacitation stage , colocalizing with tyrosine phosphorylated proteins in the acrosome and midpiece region of capacitated boar sperm . This association was confirmed by coimmunoprecipitation analysis . We also showed that bromocriptine ( 100 nM ) and low-concentration dopamine ( 100 nM and 10 muM ) increased total and progressive motility of sperm . However , high concentrations of dopamine ( 1 mM ) decreased tyrosine phosphorylation and motility in in vitro sperm capacitation assays . This can be explained by the presence of the dopamine transporters ( Q01959 REA , official symbol Q01959 REA ) in sperm , as demonstrated by Western blot analysis and immunocytochemistry . Taken together , our results support the idea that dopamine may have a fundamental role during sperm capacitation and motility in situ in the female upper reproductive tract .

Optimizing management of treatment-naïve and treatment-experienced HIV + patients : the role of maraviroc . DB04835 MEN is the first P51681 REA antagonist approved for the treatment of HIV - 1 infection . It specifically inhibits the replication of R5 viruses by blocking viral entry . HIV - 1 tropism can be estimated accurately and predict viral response to maraviroc . Genotypic tools are increasingly replacing phenotypic assays in most places . The favorable pharmacokinetic properties and the good safety profile of maraviroc may support an earlier use of the drug in HIV - 1 infection , as well as favor its consideration as part of switch strategies in patients under suppressive antiret-roviral regimens containing less-well-tolerated drugs . Moreover , a particular immune benefit of maraviroc might encourage its use as part of intensification strategies in HIV-infected patients with impaired P01730 REA gains despite prolonged suppression of HIV replication with antiretroviral therapy . However , the long-term consequences of using maraviroc must be carefully checked , given its particular mechanism of action , blocking a physiologic cell receptor .

Dissociable fronto-striatal effects of dopamine D2 receptor stimulation on cognitive versus motor flexibility . Genetic and pharmacological studies suggest an important role of the dopamine D2 receptor ( P14416 REA ) in flexible behavioral adaptation , mostly shown in reward-based learning paradigms . Recent evidence from imaging genetics indicates that also intentional cognitive flexibility , associated with lateral frontal cortex , is affected by variations in P14416 REA signaling . In the present functional magnetic resonance imaging ( Q9BWK5 ) study , we tested the effects of a direct pharmacological manipulation of P14416 REA stimulation on intentional flexibility in a task-switching context , requiring switches between cognitive task rules and between response hands . In a double blind , counterbalanced design , participants received either a low dose of the P14416 REA agonist bromocriptine or a placebo in two separate sessions . DB01200 SUB modulated the blood-oxygen-level-dependent ( BOLD ) signal during rule switching : rule-switching-related activity in the left posterior lateral frontal cortex and in the striatum was increased compared to placebo , at comparable performance levels . Fronto-striatal connectivity under bromocriptine was slightly increased for rule switches compared to rule repetitions . Hand-switching-related activity , in contrast , was reduced under bromocriptine in sensorimotor regions . Our results provide converging evidence for an involvement of P14416 REA signaling in fronto-striatal mechanisms underlying intentional flexibility , and indicate that the neural mechanisms underlying different types of flexibility ( cognitive vs motor ) are affected differently by increased dopaminergic stimulation .

Neuronal ablation of p-Akt at Ser 473 leads to altered P08908 REA / 2A receptor function . The serotonergic system regulates a wide range of behavior , including mood and impulsivity , and its dysregulation has been associated with mood disorders , autism spectrum disorder , and addiction . Diabetes is a risk factor for these conditions . P01308 REA resistance in the brain is specifically associated with susceptibility to psychostimulant abuse . Here , we examined whether phosphorylation of Akt , a key regulator of the insulin signaling pathway , controls serotonin ( 5 - HT ) signaling . To explore how impairment in Akt function regulates 5 - HT homeostasis , we used a brain-specific rictor knockout ( KO ) mouse model of impaired neuronal phosphorylation of Akt at Ser 473 . Cortical P08908 REA and 5 - Q13049 REA receptor binding was significantly elevated in rictor KO mice . Concomitant with this elevated receptor expression , the P08908 REA receptor agonist 8 - Hydroxy - 2 - ( di-n-propylamino ) tetralin ( 8 - OH-DPAT ) led to an increased hypothermic response in rictor KO mice . The increased cortical P08908 REA receptor density was associated with higher P08908 REA receptor levels on the cortical cell surface . In contrast , rictor KO mice displayed significantly reduced head-twitch response ( HTR ) to the 5 - Q13049 REA / C agonist 2,5- dimethoxy - 4 - iodoamphetamine ( DOI ) , with evidence of impaired 5 - Q13049 REA / C receptor signaling . In vitro , pharmacological inhibition of Akt significantly increased P08908 REA receptor expression and attenuated DOI-induced 5 - Q13049 REA receptor signaling , thereby lending credence to the observed in vivo cross-talk between neuronal Akt signaling and 5 - HT receptor regulation . These data reveal that defective central Akt function alters 5 - HT signaling as well as 5 - HT-associated behaviors , demonstrating a novel role for Akt in maintaining neuronal 5 - HT receptor function .

Inhibition of Akt / P31749 REA by a P35354 REA inhibitor induces apoptosis in gastric cancer cells . BACKGROUND / AIM : Inhibition of cyclooxygenase - 2 has been proposed to be a potential mechanism for the chemoprevention of gastrointestinal tumors by nonsteroidal anti-inflammatory drugs . This study investigates the mechanisms by which the cyclooxygenase - 2 inhibitor SC236 induces apoptosis of gastric cancer cell lines and its downstream signaling pathway . METHODS : Two gastric cancer cell lines , AGS and MKN 28 , were treated with SC236 and assessed for cell growth and apoptosis . The involvement of mitogen-activated protein kinase and Akt kinase / protein kinase B ( Akt / P31749 REA ) pathways and their downstream signalings were studied in the AGS cell line . RESULTS : SC236 treatment induced apoptosis in gastric cancer cells and caused activation of p38 and stress-activated protein kinase / jun kinase , but down-regulated Akt / P31749 REA . The specific p38 inhibitor SB203580 and the dominant-negative stress-activated protein kinase / jun kinase both failed , while the constitutively active form of Akt / P31749 REA was able to block SC236 - induced apoptosis . SC236 - induced apoptosis was coupled with release of cytochrome c and activation of caspases . CONCLUSION : One of the pathways involved in SC - 236 - induced apoptosis in gastric cancer cells is through downregulation of Akt and then release of cytochrome c .

A 3 - D model for P08908 REA - receptor agonists based on stereoselective methyl-substituted and conformationally restricted analogues of 8 - hydroxy - 2 - ( dipropylamino ) tetralin . The enantiomers of cis - and trans -1,2 , 3,4 , 4a , 5,10 , 10a - octahydro - 9 - hydroxy - 1 - propylbenzo [ g ] quinolines ( 10 and 11 , respectively ) and the enantiomers of trans -1,2 , 3,4 , 4a , 5,6 , 10b - octahydro - 10 - hydroxy - 4 - propylbenzo [ f ] quinoline ( 12 ) have been synthesized and their stereochemical and conformational characteristics have been studied by use of X-ray crystallography and molecular mechanics ( P08253 REA ) calculations . The compounds , which are conformationally restricted analogues of the potent 5 - hydroxytryptamine ( 5 - HT ) receptor agonist 8 - hydroxy - 2 - ( dipropylamino ) tetralin ( 8 - OH-DPAT ; 1 ) have been evaluated for central 5 - HT and dopamine receptor stimulating activity by use of biochemical and behavioral tests in rats . In addition , we have evaluated the ability of these compounds and a number of previously reported analogues to displace [ 3H ] - 8 - OH-DPAT from P08908 REA - binding sites . The enantiomers of 12 behave as potent P08908 REA - receptor agonists , whereas the octahydrobenzo [ g ] quinoline derivatives are much less potent or inactive . In general , the affinities of the compounds correlate well with their agonist potencies . The set of compounds under study is accommodated by a novel computer-graphics-derived model for P08908 REA - receptor agonism . The model consists of a flexible pharmacophore and a partial receptor-excluded volume .

Agonism at P41595 REA receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5 - hydroxytryptamine 2B ( P41595 REA ) receptors . To evaluate whether agonism at P41595 REA receptors is a phenomenon of the class of the ergolines , we studied P41595 REA receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC 50 8.42 and 8.72 ) . DB01200 SUB acted as a partial agonist ( pEC 50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5 - HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 REA receptors seems not to be a class effect of the ergolines .

P01308 REA action on H292 bronchial carcinoma cells as compared to normal bronchial epithelial cells . DB00030 MENMAX DB00030 MEN may contribute to bronchial carcinoma due to P08069 REA activation by high local concentrations . Therefore , effects of insulin and P05019 REA on human bronchial carcinoma cells ( H292 ) and normal bronchial epithelium cells ( P02100 REA ) were studied . TGF-β was included since it also influences carcinoma progression . H292 and P02100 REA cells expressed both the insulin receptor and the P08069 REA ; in H292 cells an additional , shorter , splicing variant ( IR-A ) of the insulin receptor was present . P06213 REA expression was around four to five times higher in H292 than in P02100 REA cells at mRNA and protein levels . P01308 REA and TGF-β exerted contrary actions on proliferation and gene expression in H292 cells . Genes regulated by insulin , P05019 REA , and TGF-β were linked to inflammation , cell adhesion , muscle contraction and differentiation . P01308 REA and P05019 REA also suppressed DNA repair genes . EC ( 50 ) for insulin-induced proliferation was around 5 nM in H292 and around 30 nM P02100 REA cells . The EC ( 50 ) values for gene expression ranged from 9 to 90 nM in both cell types , dependent on the gene studied . In H292 cells , the proliferative response was much stronger if TGF-β was present . In P02100 REA cells this interaction of insulin and TGF-β was not observed , and changes in gene expression were mostly lower by at least 10 - fold as compared to H292 . All in all , the insulin effects in H292 were generally much stronger than in P02100 REA cells and - with regard to proliferation - occurred at lower concentrations . Thus , insulin will hardly induce cancer from normal bronchial cells but may favour progression of pre-existing tumours .

Antimetastatic potential of P05121 REA - specific RNA aptamers . The serine protease inhibitor plasminogen activator inhibitor - 1 ( P05121 REA ) is increased in several cancers , including breast , where it is associated with a poor outcome . Metastatic breast cancer has a dismal prognosis , as evidenced by treatment goals that are no longer curative but are largely palliative in nature . P05121 REA competes with integrins and the urokinase plasminogen activator receptor on the surface of breast cancer cells for binding to vitronectin . This results in the detachment of tumor cells from the extracellular matrix , which is critical to the metastatic process . For this reason , we sought to isolate RNA aptamers that disrupt the interaction between P05121 REA and vitronectin . Through utilization of combinatorial chemistry techniques , aptamers have been selected that bind to P05121 REA with high affinity and specificity . We identified two aptamers , WT - 15 and Q9GZT9 , that disrupt the interactions between P05121 REA and heparin , as well as P05121 REA and vitronectin , without affecting the antiprotease activity of P05121 REA . Furthermore , Q9GZT9 prevented the detachment of breast cancer cells ( MDA-MB - 231 ) from vitronectin in the presence of P05121 REA , resulting in an increase in cellular adhesion . Therefore , the P05121 REA aptamer Q9GZT9 demonstrates therapeutic potential as an antimetastatic agent and could possibly be used as an adjuvant to traditional chemotherapy for breast cancer .

A selenosemicarbazone complex with copper efficiently down-regulates the 90 - kDa heat shock protein P07900 REA and its client proteins in cancer cells . BACKGROUND : The 90 - kDa heat shock protein P07900 REA is critical for the stability of several proteins that are important for tumor progression and thus , is a promising target for cancer therapy . Selenosemicarbazone metal complexes have been shown to possess anticancer activity through an unknown molecular mechanism . METHODS : The MTT assay , fluorescence-activated cell sorting , and fluorescent microscopy were used to analyze the mechanism of the anti-cancer activity of the selenosemicarbazone metal complexes . Additionally , RNA-seq was applied to identify transcriptional gene changes , and in turn , the signaling pathways involved in the process of 2-24 a / Cu-induced cell death . Last , the expression of P07900 REA , P0DMV8 , P11309 REA , and AKT proteins in 2-24 a / Cu-treated cells were investigated by western blot analysis . RESULTS : A novel selenosemicarbazone copper complex ( 2-24 a / Cu ) efficiently induced G2 / M arrest and was cytotoxic in cancer cells . 2-24 a / Cu significantly induced oxidative stress in cancer cells . Interestingly , although RNA-seq revealed that the transcription of P07900 REA was increased in 2-24 a / Cu-treated cells , western blotting showed that the expression of P07900 REA protein was significantly decreased in these cells . Furthermore , down-regulation of P07900 REA led to the degradation of its client proteins ( P11309 REA and P31749 REA ) , which are also cancer therapy targets . CONCLUSION : Our results showed that 2-24 a / Cu efficiently generates oxidative stress and down-regulates P07900 REA and its client proteins ( P11309 REA , P31749 REA ) in U2os and HeLa cells . These results demonstrate the potential application of this novel copper complex in cancer therapy .

Co-regulation between cyclo-oxygenase - 2 and inducible nitric oxide synthase expression in the time-course of murine inflammation . Many in vitro studies have used cell cultures to focus on the relationships between cyclo-oxygenase - 2 ( P35354 REA ) and inducible nitric oxide synthase ( P35228 REA ) isoforms . We have investigated the time-course of regulation and the role of P35354 REA and P35228 REA in a model of experimental inflammation in mice , the air pouch injected with zymosan . This study demonstrates that there is an early acute phase ( 4 h ) mediated mainly by eicosanoids , with high levels of prostaglandin E2 ( DB00917 ) produced by cyclo-oxygenase - 1 . In addition , in the later phase ( from 12 h ) there is a participation of nitric oxide ( NO ) and DB00917 accompanied by co-induction of both P35228 REA and P35354 REA . These enzymes were detected in migrating leukocytes as well as in macrophages lining the air pouch . Administration of NS398 or indomethacin inhibited DB00917 levels and P36551 REA activity , but also nitrite levels and P35228 REA activity , which was accompanied by a reduction in P35228 REA expression . DB02533 MEN inhibited nitrite levels and P35228 REA activity in addition to exerting inhibitory effects on the P36551 REA pathway . Treatment of animals with dexamethasone reduced nitrite and DB00917 concentrations in air pouch exudates , as well as P35228 REA and P35354 REA expression in migrating cells . Our results indicate that DB00917 and NO may play in vivo mutual modulatory roles in the inflammatory response caused by zymosan injection into the mouse air pouch , a suitable model to study drugs acting on those pathways .

Development of peptidomimetic ligands of Pro - DB00149 - DB00145 - NH ( 2 ) as allosteric modulators of the dopamine D ( 2 ) receptor . A variety of stable , small-molecule peptidomimetic ligands have been developed to elucidate the mechanism by which the neuropeptide Pro - DB00149 - DB00145 - NH ( 2 ) ( P00747 REA ) modulates dopaminergic neurotransmission . Photoaffinity labeling ligands based upon P00747 REA peptidomimetics have been used to establish that P00747 REA binds to the P14416 REA at a site that is different from the orthosteric site , thus making P00747 REA and its peptidomimetics allosteric modulators of the dopamine receptor . Through the design , synthesis and pharmacological evaluation of conformationally constrained peptidomimetics containing lactam , bicyclic , and spiro-bicyclic scaffolds , support was provided for the hypothesis that the bioactive conformation of P00747 REA is a type II β-turn . In addition , studies with peptidomimetics designed to mimic either a type VI β-turn or polyproline II helix conformation yielded molecules that were able to modulate dopamine receptors because of their ability to place the carboxamide NH ( 2 ) pharmacophore in the same topological space as that seen in the type II β-turn . Extensive studies with the spiro-bicyclic P00747 REA peptidomimetics also established that both positive and negative modes of modulation were possible for the same series of peptidomimetics simply as a result of minor differences in the stereochemistry about the bridgehead carbon within the scaffold . This information was used to transform existing positive modulators into negative modulators , which demonstrated that small structural changes in the spiro-bicyclic dopamine receptor modulators are capable of causing major changes in the modulatory activity of P00747 REA peptidomimetics .

[ DB02424 MEN administration reduces the number of P07900 REA - positive germ cells in the mouse embryo : preliminary results ] . 5 mg of DB02424 MEN , an inhibitor of stress protein P07900 REA which express on mammalian germ cells , were administered to E8 pregnant mice . E17 embryos were removed , and a quantitative analysis of HSP 90 - immunoreactive cells in the gonad was performed , in comparison to control embryos . First , we observed that the number of germ cells is lower in male than in female embryos , as well in control and experimental embryos . External features of experimental and control embryos did not display any difference . Embryos exposed to geldanamycin exhibit a significant decrease of immunoreactive germ cells . In two embryos , we observed a group of ectopic immunoreactive cells in the pelvic area . We conclude that geldanamycin inhibits germ cells migration , and suggest that this inhibition can lead to ectopic germ cell populations , similar to teratomas .

Protease inhibitors prevent plasminogen-mediated , but not pemphigus vulgaris-induced , acantholysis in human epidermis . Pemphigus is an autoimmune blistering disease of the skin and mucous membranes . It is caused by autoantibodies directed against desmosomes , which are the principal adhesion structures between epidermal keratinocytes . Binding of autoantibodies leads to the destruction of desmosomes resulting in the loss of cell-cell adhesion ( acantholysis ) and epidermal blisters . The plasminogen activator system has been implicated as a proteolytic effector in pemphigus . We have tested inhibitors of the plasminogen activator system with regard to their potential to prevent pemphigus-induced cutaneous pathology . In a human split skin culture system , IgG preparations of sera from pemphigus vulgaris patients caused histopathologic changes ( acantholysis ) similar to those observed in the original pemphigus disease . All inhibitors that were tested ( active site inhibitors directed against uPA , tPA , and / or plasmin ; antibodies neutralizing the enzymatic activity of uPA or tPA ; substances interfering with the binding of uPA to its specific cell surface receptor Q03405 REA ) failed to prevent pemphigus vulgaris IgG-mediated acantholysis . P00747 REA - mediated acantholysis , however , was effectively antagonized by the synthetic active site serine protease inhibitor DB05476 MEN or by p-aminomethylbenzoic acid . Our data argue against applying anti-plasminogen activator / anti-plasmin strategies in the management of pemphigus .

DB00126 MEN is dispensable for oxygen sensing in vivo . Prolyl - 4 - hydroxylation is necessary for proper structural assembly of collagens and oxygen-dependent protein stability of hypoxia-inducible transcription factors ( HIFs ) . In vitro function of HIF prolyl - 4 - hydroxylase domain ( P20941 ) enzymes requires oxygen and 2 - oxoglutarate as cosubstrates with iron ( II ) and vitamin C serving as cofactors . Although vitamin C deficiency is known to cause the collagen-disassembly disease scurvy , it is unclear whether cellular oxygen sensing is similarly affected . Here , we report that vitamin C-deprived Gulo ( - / - ) knockout mice show normal HIF-dependent gene expression . The systemic response of Gulo ( - / - ) animals to inspiratory hypoxia , as measured by plasma erythropoietin levels , was similar to that of animals supplemented with vitamin C . Hypoxic HIF induction was also essentially normal under serum - and vitamin C-free cell-culture conditions , suggesting that vitamin C is not required for oxygen sensing in vivo . Glutathione was found to fully substitute for vitamin C requirement of all 3 P20941 isoforms in vitro . Consistently , glutathione also reduced HIF - 1α protein levels , transactivation activity , and endogenous target gene expression in cells exposed to CoCl ( 2 ) . A Cys 201Ser mutation in Q9GZT9 increased basal hydroxylation rates and conferred resistance to oxidative damage in vitro , suggesting that this surface-accessible Q9GZT9 cysteine residue is a target of antioxidative protection by vitamin C and glutathione .