MH_dev_315

Update on the genetics characterization of vitiligo . Vitiligo is an autoimmune skin disorder in which autoimmune-mediated destruction of melanocytes caused depigmentation of skin patches . The complex genetics of vitiligo involves multiple susceptibility loci , genetic heterogeneity and incomplete penetrance with gene-gene and gene-environment interactions . In order to clarify the genetic factors , two different principal approaches have applied for the identification of genomic regions or candidate genes that mediate susceptibility to vitiligo . First approach is the genome-wide linkage analyses , which is conducted by scanning of entire human genome for genomic regions that are linked to the development of vitiligo . The other approach is functional candidate gene association ( FCGA ) analyses that detect specific candidate genes , which are expected to involve in disease on the basis of their priori biological functions . Genomic-wide scans have provided a strong support for vitiligo susceptibility genes on chromosomes 4q13 - q21 , 1p31 , 7q22 , 8p 12 and 17p13 , while loci of interest at 6p , 6q , 14q , 9q , 13q , 19p and 22q required further follow-up . Whereas , FCGA studies have identified some candidate genes which are associated with vitiligo , such as HLA , O43918 , Q86XK2 , CAT , Q9UJU5 , P03372 REA , P21964 REA , Q9Y2R2 REA , Q9C000 REA , P16234 REA , MYG 1 , O75030 REA , CD117 , P17861 REA , FAS , P35354 REA , P05305 REA and P12821 REA , but few of them reports now appear to be false-positive . This review will provides an update on genetics of vitiligo based on the identification of novel candidate genes that represent , in my opinion as optimal utility for future therapeutic targets in the pathogenesis of vitiligo .

P26441 REA and interleukin - 6 differentially activate microglia . Studies have shown that cytokines released following CNS injury can affect the supportive or cytotoxic functions of microglia . P05231 REA ( P05231 REA ) - family cytokines are among the injury factors released . To understand how microglia respond to P05231 REA family cytokines , we examined the effects of ciliary neurotrophic factor ( P26441 REA ) and P05231 REA on primary cultures of rat microglia . To assess the functional state of the cells , we assayed the expression of tumor necrosis factor-alpha ( TNFalpha ) , interleukin - 1beta ( IL - 1beta ) , and cyclooxygenase 2 ( P35354 REA ) following stimulation . We show that P26441 REA reduces P35354 REA levels , whereas P05231 REA increases the expression of IL - 1beta , TNFalpha , and Cox - 2 . We also examined trophic factor expression and found that P26441 REA enhances glial cell-line derived neurotrophic factor ( P39905 REA ) mRNA and protein secretion , whereas P05231 REA has no effect . Correspondingly , conditioned media from P26441 REA - stimulated microglia promote motor neuron survival threefold beyond controls , whereas P05231 REA - stimulated microglia decrease neuronal survival twofold . To understand better the signaling mechanisms responsible for the opposite responses of these P05231 REA - family cytokines , we examined P35610 - 3 and P29323 REA phosphorylation in P26441 REA - and P05231 REA - stimulated microglia . P05231 REA markedly increases P35610 - 3 and P29323 REA phosphorylation after 20 min of treatment , whereas these signal transducers are weakly stimulated by P26441 REA across a range of doses . We conclude that P26441 REA modifies microglial activation to support neuronal survival and that P05231 REA enhances their capacity to do harm , as a result of different modes of intracellular signaling .

DB00472 SUB - induced proliferation and differentiation of neural progenitor cells isolated from rat postnatal cerebellum . Previous studies have shown that the serotonin-reuptake inhibitor ( SSRI ) fluoxetine affects neural progenitors derived from postnatal cerebellum or adult hippocampus and stimulates their proliferation . In the human cerebellum , the proliferation of cerebellar granule cells ( CGC ) continues until the 11th postnatal month and could be influenced in infants by breastfeeding-delivered SSRIs . Current information about fluoxetine effects on postnatal cerebellar neural progenitors is limited . Here we report the characterization of fluoxetine actions on rat postnatal cerebellar neural progenitors . RT-PCR and immunostaining revealed the expression of serotonin transporter ( P31645 REA ) , 5HT ( 1A ) receptors , tryptophan hydroxylase ( P17752 REA ) , and serotonin ( 5HT ) . Protracted in vitro fluoxetine treatment increased cell proliferation and differentiation . The proliferative effects of fluoxetine , 5HT , and the selective agonist of 5HT ( 1A ) receptors trans - 8 - hydroxy - 2 - ( N-n-propyl-N - 3 ' - iodo - 2 ' - propenyl ) aminotetralin ( 8 - OH-PIPAT ) were abolished by the selective antagonist of 5HT ( 1A ) receptors , N - [ 2 - [ 4 - ( 2 - methoxyphenyl ) - 1 - piperazinyl ] ethyl ] - N - ( 2 - pyridinyl ) cyclohexanecarboxamide trihydrochloride ( WAY - 100635 ) . Furthermore , fluoxetine-induced activation of both the DB02527 - response element-binding ( CREB ) protein and extracellular signal-regulated protein kinases ( P27361 REA / 2 ) , which was abolished by the selective inhibitor of Q96HU1 kinase kinase ( MEK ) 1,4- diamino -2,3- dicyano -1,4- bis ( 2 - aminophenylthio ) butadiene ( U0126 ) , and increased cyclin D1 expression . All these effects were prevented by WAY - 100635 . Collectively , our results demonstrate that rat postnatal cerebellum contains neural progenitors capable of proliferating and differentiating in response to fluoxetine exposure , possibly through the activation of 5HT ( 1A ) receptors . The relevance of these findings for possible SSRI effects on the developing postnatal / infant human cerebellum should be explored .

DB01221 receptor modulation by the neuropeptide apelin : implications for excitotoxic injury . Excitotoxic neuronal damage via over-activation of the DB01221 receptor has been implicated in many neurodegenerative diseases . In vitro modeling of excitotoxic injury has shown that activation of G-protein coupled receptors ( GPCRs ) counteracts such injury through modulation of neuronal pro-survival pathways and / or DB01221 receptor signaling . We have previously demonstrated that the GPCR P35414 REA and its endogenous neuropeptide ligand apelin can protect neurons against excitotoxicity , but the mechanism ( s ) of this neuroprotection remain incompletely understood . We hypothesized that apelin can promote neuronal survival by activating pro-survival signaling as well as inhibiting DB01221 receptor-mediated excitotoxic signaling cascades . Our results demonstrate that ( i ) apelin activates pro-survival signaling via inositol trisphosphate ( IP ( 3 ) ) , protein kinase C ( PKC ) , mitogen-activated protein kinase kinase 1/2 ( Q02750 REA / 2 ) , and extracellular signal-regulated kinase -1/2 ( P27361 REA / 2 ) to protect against excitotoxicity , and ( ii ) apelin inhibits excitotoxic signaling by attenuating DB01221 receptor and calpain activity , and by modulating DB01221 receptor subunit Q13224 REA phosphorylation at serine 1480 . These studies delineate a novel apelinergic signaling pathway that concurrently promotes survival and limits DB01221 receptor-mediated injury to protect neurons against excitotoxicity . Defining apelin-mediated neuroprotection advances our understanding of neuroprotective pathways and will potentially improve our ability to develop therapeutics for excitotoxicity-associated neurodegenerative disorders .

Pathogenesis of spinally mediated hyperalgesia in diabetes . Hyperalgesia to noxious stimuli is accompanied by increased spinal cyclooxygenase ( P36551 REA ) - 2 protein in diabetic rats . The present studies were initiated to establish causality between increased spinal P35354 REA activity and hyperalgesia during diabetes and to assess the potential involvement of polyol pathway activity in the pathogenesis of spinally mediated hyperalgesia . Rats with 1 , 2 , or 4 weeks of streptozotocin-induced diabetes exhibited significantly increased levels of spinal P35354 REA protein and activity , along with exaggerated paw flinching in response to 0.5 % paw formalin injection . Increased flinching of diabetic rats was attenuated by intrathecal pretreatment with a selective P35354 REA inhibitor immediately before formalin injection , confirming the involvement of P35354 REA activity in diabetic hyperalgesia . Chronic treatment with insulin or ICI 222155 , an aldose reductase inhibitor ( Q9Y4X5 REA ) previously shown to prevent spinal polyol accumulation and formalin-evoked hyperalgesia in diabetic rats , prevented elevated spinal P35354 REA protein and activity in diabetic rats . In contrast , the Q9Y4X5 REA IDD 676 had no effect on spinal polyol accumulation , elevated spinal P35354 REA , or hyperalgesia to paw formalin injection . In the spinal cord , aldose reductase immunoreactivity was present solely in oligodendrocytes , which also contained P35354 REA immunoreactivity . Polyol pathway flux in spinal oligodendrocytes provides a pathogenic mechanism linking hyperglycemia to hyperalgesia in diabetic rats .

Dual ligands targeting dopamine D2 and serotonin P08908 REA receptors as new antipsychotical or anti-Parkinsonian agents . Psychiatric disorders like schizophrenia and neurodegenerative diseases like Parkinson ' s disease are associated with poly-factorial pathogenic mechanisms , with several neurotransmitter systems closely involved . In addition to the cerebral dopaminergic ( DA ) system , the serotoninergic ( 5 - HT ) system also plays a crucial role in regulating psychoemotional , cognitive and motor functions in the central nervous system ( CNS ) . Among the large 5 - HT receptor family , accumulating data have revealed new insights into the therapeutic benefit of the P08908 REA receptor in treating various CNS disorders , especially schizophrenia and Parkinson ' s disease . The present review discusses the advance of dual agents with mixed actions at the dopamine D2 and serotonin P08908 REA receptors in the treatment of these diseases . Aripiprazole was the only marketed drug with dual D2 and P08908 REA profile . It is a partial D2 and P08908 REA receptor agonist and has been prescribed as an atypical antipsychotical drug . Two other drugs DB06016 and Pardoprunox are being investigated in clinic . Most of the other candidate compounds , including DB04888 MEN , Sarizotan , Mazapertine succinate , PF - 217830 , and Adoprazine were discontinued due to either non-optimal pharmacokinetic properties or insufficient therapeutical efficacy . Although much effort has been done to highlight the advantages of the P08908 REA and D2 dual approach , it has to be pointed out that many of these drugs showed poly-pharmacological profile by targeting many other receptors and / or transporters besides the D2 and P08908 REA receptors . In this regard , ' pure ' compounds exclusively acting on the D2 and P08908 REA receptors are highly needed to further validate this approach . Meanwhile , safety concerns and in vivo pharmacokinetic alerts should also be implanted to the drug design art early .

P13639 REA mutants deficient in diphthamide formation show temperature-sensitive cell growth . Protein synthesis elongation factor 2 ( P13639 REA ) from eukaryotes contains an unusual modified histidine residue , termed diphthamide . DB03223 MEN has been shown to be a site of ADP-ribosylation by bacterial toxins , but its function remains obscure . We expressed mutant genes of P13639 REA with substitutions of 19 other amino acids for DB00117 - 699 , which is modified to diphthamide , in yeast cells and found that they can be classified into three groups . In the first group ( Group 1 ) , replacement of DB00117 - 699 by the basic amino acid DB00125 or Lys showed not only loss of P13639 REA activity but also inhibitory effects on the growth of cells co-expressing wild-type P13639 REA . In the second group ( Group 2 ) , replacement with DB00145 , Pro , DB00133 , or DB00128 resulted in nonfunctional P13639 REA , but it did not affect the growth of cells co-expressing wild-type P13639 REA . In the third group ( Group 3 ) , replacement by one of the other 13 amino acids resulted in a functional P13639 REA . In the Group 3 mutants , P13639 REA was not ribosylated by diphtheria toxin , indicating that the mutant EF - 2s did not form diphthamide . However , the viable cells grew more slowly than cells expressing wild-type P13639 REA and showed temperature sensitivities . This result suggests that diphthamide may confer heat resistance on P13639 REA , although it still may be active without diphthamide at a normal temperature .

Q9Y2D1 REA polymorphisms influence P39905 REA function and response to treatment in children with childhood acute lymphoblastic leukemia . DB00023 is a standard and critical component in the therapy of childhood acute lymphoblastic leukemia . DB00174 MEN synthetase ( P08243 REA ) and the basic region leucine zipper activating transcription factor 5 ( Q9Y2D1 REA ) and arginosuccinate synthase 1 ( P00966 REA ) have been shown to mediate the antileukemic effect of asparaginase and to display variable expression between leukemia cells that are resistant and sensitive to treatment . Fourteen polymorphisms in the regulatory and coding regions of these genes were investigated for an association with acute lymphoblastic leukemia outcome . Lower event-free survival ( O43281 ) was associated with Q9Y2D1 REA T1562C , tandem-repeat P08243 REA polymorphism , derived haplotype , and P00966 REA G1343T and G34T substitutions ( P ≤ . 03 ) . Associations were limited to patients who received Escherichia coli asparaginase . Variations that sustained correction for multiple testing ( Q9Y2D1 REA T1562C , P = . 005 ; P08243 REA tandem-repeat and related haplotype , P ≤ . 01 ) were subsequently analyzed in the replication cohort . The E coli-dependent association of the Q9Y2D1 REA T1562 allele with reduced O43281 was confirmed ( P = . 01 ) . A gene-reporter assay showed that the haplotype tagged by T1562 had higher promoter activity ( P ≤ . 01 ) . The remaining regulatory polymorphisms also appeared to affect Q9Y2D1 REA function ; 2 additional high-activity haplotypes were identified ( P ≤ . 02 ) and were further corroborated by quantitative mRNA analysis in lymphoblastoid cell lines . The Q9Y2D1 REA - regulated increase in P08243 REA expression in response to more efficacious E coli-induced asparagine depletion may explain our observed results .

Lack of biological relevance of platelet cyclooxygenase - 2 dependent thromboxane A2 production . INTRODUCTION : There is emerging evidence of a considerable variability of the impact of aspirin on clinical outcome and laboratory findings . Persistent TxA 2 production seems to be the most likely reason . Aim of this study was to determine whether the mechanism responsible for TxA 2 persistent production is , at least partially , dependent upon aspirin-insensitive platelet P35354 REA enzymatic pathway . METHODS AND RESULTS : In 100 consecutive patients , under chronic aspirin anti-platelet treatment ( 100-160 mg / day ) selected on the basis of detectable plasma salicylate levels , serum and DB04557 MEN ( AA ) - induced platelet TxA 2 production , immunoblot analysis of platelet P23219 REA / P35354 REA expression and P35354 REA activity were studied . Immunoblot revealed P35354 REA expression in 46 % patients , in an amount that was markedly lower than P23219 REA . In 10 P35354 REA positive patients with TxA 2 levels over the median , AA-induced TxA 2 production performed in vitro in the presence of the P35354 REA inhibitor CAY 10404 and aspirin demonstrated that P35354 REA dependent TxA 2 production is less than 2 % . CONCLUSION : Our data demonstrate that the inter-individual variability of platelet sensitivity to aspirin is due to a reduced efficacy of aspirin on platelet P23219 REA despite ascertained patient compliance . We suggest that serum TxA 2 assay might be performed in future clinical studies to improve our knowledge on the residual TxA 2 production in aspirin-treated patients .

Transcriptional profiles during the differentiation and maturation of monocyte-derived dendritic cells , analyzed using focused microarrays . Dendritic cells ( DC ) are professional antigen-presenting cells capable of initiating primary immune responses . They have been intensively studied and are used in both basic immunology research and clinical immunotherapy . However , the genetic pathways leading to DC differentiation and maturation remain poorly understood . Using focused microarrays with oligonucletotide probes for 120 genes encoding co-stimulatory molecules , chemokines , chemokine receptors , cytokines , cytokine receptors , TLRs , and several other related molecules , we analyzed the kinetics of gene expression for the overall differentiation process of monocytes into mature DC . In parallel , we compared the transcriptional profiles in DC maturation in the presence of LPS , P01375 REA or trimeric P29965 REA . We found similar transcriptional profiles for early immature DC and immature DC , respectively generated by culturing monocytes with GM - P04141 REA and P05112 REA for three or six days . We identified sets of common and stimuli-specific genes , the expression of which changed following stimulation with LPS , P01375 REA or P29965 REA . A dynamic analysis of the entire DC differentiation and maturation process showed that some important inflammatory and constitutive chemokines are transcribed in both immature and mature DC . The correlative expression kinetics of the gene pairs P14778 REA / P27930 REA , P40933 REA / Q13261 REA , Q9NNX6 / P13598 REA and Q9NNX6 / P32942 REA imply that they all play crucial roles in mediating DC functions . Thus , our analysis with focused microarrays shed light on the transcriptional kinetics of DC differentiation and maturation , and this method may also prove useful for identifying novel marker genes involved in DC functions .

P40933 REA affects serotonin system and exerts antidepressive effects through IL15Rα receptor . Contrary to the reduction of depressive-like behavior observed in several strains of cytokine receptor knockout mice , mice lacking the specific receptor for interleukin ( IL ) - 15 showed increased immobility in tail suspension and modified forced swimming tests . There was also a reduction in social interactions . The hippocampus of the IL15Rα knockout mice had decreased mRNA for 5 - HT ( 1A ) , increased mRNA for 5 - HT ( 2C ) , and region-specific changes of serotonin reuptake transporter ( P31645 REA ) immunoreactivity . DB00472 SUB ( the classic antidepressant DB00472 SUB , which inhibits 5 - HT ( 2C ) and P31645 REA ) reduced the immobility of the IL15Rα knockout mice in comparison with their pretreatment baseline . Together with the unchanged performance of the IL15Rα knockout mice on the rotarod , this response to fluoxetine indicates that the immobility reflects depression . Wildtype mice responded to P40933 REA treatment with improvement of immobility induced by forced swimming , whereas the knockout mice failed to respond . Thus , the cognate P40933 REA receptor is necessary for the antidepressive activity of P40933 REA . In ex vivo studies , P40933 REA decreased synaptosomal uptake of 5 - HT , and modulated the expression of 5 - HT ( 2C ) and P31645 REA in cultured neurons in a dose - and time-dependent manner . Thus , the effect of P40933 REA on serotonin transmission may underlie the depressive-like behavior of IL15Rα knockout mice . We speculate that P40933 REA is essential to maintain neurochemical homeostasis and thereby plays a role in preventing neuropsychiatric symptoms .

DB00472 SUB induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 SUB ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5 - HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 REA ) , and cyclooxygenase - 2 ( P35354 REA ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg ( - 1 ) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 REA ; i . p . , 125mgkg ( - 1 ) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5 - HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 REA mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5 - hydroxyindoleacetic acid ( 5 - HIAA ) levels ( P < 0.01 ) and , P28335 REA receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 REA ( P < 0.001 ) , and P35354 REA expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5 - HT metabolism and / or its ability to reduce colonic malignant events .

[ Effects of chronic fluoxetine treatment on catalepsy and immune response in mice genetically predisposed to freezing reaction : the role of P08908 REA and 5 - Q13049 REA receptors and tph 2 and P31645 REA genes ] . ASC / Icg ( Antidepressant Sensitive Catalepsy ) mouse strain selected for high predisposition to pinch-induced catalepsy is characterized by depressive-like behavior and impaired immune response . Chronic treatment with SSRI fluoxetine attenuated catalepsy manifestation and normalized a decreased number of rosette-forming cells ( P41440 REA ) in spleen in ASC mice . Chronic fluoxetine administration had no effect on catalepsy and P41440 REA number in mice of parental cataleptic CBA / Lac strain . DB00472 SUB failed to alter P08908 REA receptor functional activity in mice of both strains and diminished 5 - Q13049 REA receptor functional activity in CBA but not in ASC mice . No effect on cortical P08908 REA and 5 - Q13049 REA receptor mRNA levels and on P08908 REA receptor , tph 2 ( tryptophan hydroxylase - 2 ) and P31645 REA ( serotonin transporter ) mesencephalic gene expression was observed in ASC mice . Other possible serotonergic mechanisms of fluoxetine effect on catalepsy and immune response in mice with depressive-like state are discussed .

Inhibition of P51587 REA and DB01643 MEN Synthase Creates Multidrug Sensitive Tumor Cells via the Induction of Combined " Complementary Lethality " . A high mutation rate leading to tumor cell heterogeneity is a driver of malignancy in human cancers . Paradoxically , however , genomic instability can also render tumors vulnerable to therapeutic attack . Thus , targeting DNA repair may induce an intolerable level of DNA damage in tumor cells . P51587 REA mediates homologous recombination repair , and P51587 REA polymorphisms increase cancer risk . However , tumors with P51587 REA mutations respond better to chemotherapy and are associated with improved patient prognosis . P04818 REA ( TS ) is also involved in DNA maintenance and generates cellular thymidylate . We determined that antisense downregulation of P51587 REA synergistically potentiated drugs with mechanisms of action related to P51587 REA function ( cisplatin , melphalan ) , a phenomenon we named " complementary lethality . " TS knockdown induced complementary lethality to TS-targeting drugs ( 5 - FUdR and pemetrexed ) but not DNA cross-linking agents . Combined targeting of P51587 REA and TS induced complementary lethality to both DNA-damaging and TS-targeting agents , thus creating multidrug sensitive tumors . In addition , we demonstrated for the first time that simultaneous downregulation of both targets induced combined complementary lethality to multiple mechanistically different drugs in the same cell population . In this study , we propose and define the concept of " complementary lethality " and show that actively targeting P51587 REA and TS is of potential therapeutic benefit in multidrug treatment of human tumors . This work has contributed to the development of a P51587 REA - targeting antisense oligdeoxynucleotide ( ASO ) " BR - 1 " which we will test in vivo in combination with our TS-targeting ASO " Q8N1L9 83 " and attempt early clinical trials in the future.Molecular Therapy - Nucleic Acids ( 2013 ) 2 , e78 ; doi : 10.1038 / mtna . 2013.7 published online 12 March 2013 .

Plumbagin inhibits tumorigenesis and angiogenesis of ovarian cancer cells in vivo . Angiogenesis is a hallmark of tumor development and metastatic progression , and anti-angiogenic drugs targeting the P15692 REA pathway have shown to decrease the disease progression in cancer patients . In this study , we have analyzed the anti-proliferative and anti-angiogenic property of plumbagin in cisplatin sensitive , P51587 REA deficient , DB09287 - 1 and cisplatin resistant , P51587 REA proficient DB09287 - 4 ovarian cancer cells . Both DB09287 - 1 and DB09287 - 4 ovarian cancer cells are sensitive to plumbagin irrespective of P51587 REA status in both normoxia and hypoxia . Importantly , plumbagin treatment effectively inhibits P15692 REA and Glut - 1 in DB09287 - 1 and DB09287 - 4 ovarian cancer cells . We have also analyzed the p53 mutant , cisplatin resistant , and P51587 REA proficient OVCAR - 5 cells . Plumbagin challenge also restricts the P15692 REA induced pro-angiogenic signaling in HUVECs and subsequently endothelial cell proliferation . In addition , we observe a significant effect on tumor regression among OVCAR - 5 tumor-bearing mice treated with plumbagin , which is associated with significant inhibition of Ki67 and P04275 REA expressions . Plumbagin also significantly reduces CD31 expression in an ear angiogenesis assay . Collectively , our studies indicate that plumbagin , as an anti-cancer agent disrupts growth of ovarian cancer cells through the inhibition of proliferation as well as angiogenesis .

DB01088 MEN has potent anti-inflammatory properties on human monocyte-derived dendritic cells . BACKGROUND : The stable prostaglandin I2 analogue ( iloprost ) iloprost has been shown to inhibit allergic airway inflammation in mice by modulating the function of myeloid dendritic cells ( DCs ) . OBJECTIVE : The aim of the current study was to investigate the biological activity of iloprost on human monocyte-derived DCs . METHODS : I prostanoid ( IP ) receptor expression was analysed by RT-PCR . Cytokine secretion by DCs and P01730 REA + T cells was measured by ELISA . The expression of the transcription factor FoxP 3 after co-culture of DCs with P01730 REA + CD45RA + T cells was analysed by flow cytometry . RESULTS : Human monocyte-derived DCs were found to express mRNA specific for the P43119 REA IP , and stimulation with iloprost resulted in increased cyclic AMP levels in both immature DCs ( iDCs ) and mature DCs ( mDCs ) . Moreover , iloprost dose dependently inhibited the secretion of P01375 REA , P05231 REA , P10145 REA and IL - 12p70 in mDCs , while it enhanced P22301 REA production . Changes in cytokine secretion were paralleled by an altered T-cell priming capacity of DCs : in co-culture experiments of iloprost-treated mDC and naïve CD45RA + T cells , an induction of regulatory T cells could be observed , as demonstrated by increased intracellular FoxP 3 expression and P22301 REA production . Additionally , iloprost inhibited the MIP - 3beta - induced migration of mDCs . CONCLUSION : In summary , our results provide evidence that iloprost profoundly affects the function of human myeloid DCs . Therefore , iloprost might also be a new therapeutical option for the treatment of asthma in humans .

Stereospecific interaction of a novel spirosuccinimide type aldose reductase inhibitor , DB05327 MEN , with aldose reductase . P15121 REA ( AR ) is an NADPH-dependent enzyme implicated in diabetic complications . DB05327 MEN [ ( R ) - ( - ) - 2 - ( 4 - bromo - 2 - fluorobenzyl ) -1,2 , 3,4- tetrahydropyrrolo [ 1,2- a ] pyrazine - 4 - spiro - 3 ' - pyrrolidine -1,2 ' , 3,5 ' - tetrone ] is a structurally novel and potent Q9Y4X5 REA with an inhibitor constant ( K ( i ) = 10 ( - ) ( 10 ) M ) 2000 - fold lower than that of its optical antipode ( S-isomer ) . To elucidate the inhibition modes and the stereochemical differences in their inhibitory potencies , we examined the interaction of these R - and S-isomers with AR under physiological conditions . Enzyme kinetic analysis , which was performed by using physiological substrates at 37 degrees C , showed that both isomers selectively act on the E-NADP ( + ) complex in both the forward and reverse reactions of AR . However , fluorometric titration analysis demonstrated that the affinities of the isomers for the E-NADP ( + ) complex are about the same as those for the E-NADPH complex and the apoenzyme . These results suggested that the selective binding to the E-NADP ( + ) complex arises from the predominance of this enzyme form during steady-state turnover rather than from binding specificity . Both the competition with a known active site-directed Q9Y4X5 REA and the protective effect on AR inactivation by N-bromosuccinimide showed that the isomers bind to the active site of the enzyme , but the thermodynamic parameters for the binding to AR indicated that additional hydrogen bonds and / or van der Waals interactions contribute to the energetic stabilization in the E-R-isomer complex . Molecular modeling , together with the deductions from spectroscopic studies , suggested that the succinimide ring and the 4 - bromo - 2 - fluorobenzyl group of the R-isomer are optimally located for formation of a hydrogen-bonding network with AR , and that the latter benzyl group is also effective for the differentiation between AR and aldehyde reductase ( a closely related enzyme ) .

Glutamatergic regulation of the p70S6 kinase in primary mouse neurons . Brief glutamatergic stimulation of neurons from fetal mice , cultured in vitro for 6 days , activates the P42345 REA - S6 kinase , P27361 REA / 2 and Akt pathways , to an extent approaching that elicited by brain-derived neurotrophic factor . In contrast , sustained glutamatergic stimulation inhibits P29323 REA , Akt , and S6K . Glutamatergic activation of S6K is calcium / calmodulin-dependent and is prevented by inhibitors of calcium / calmodulin-dependent protein kinase 2 , phosphatidylinositol 3 - OH-kinase and by rapamycin . 2 - Amino - 5 - phosphonovaleric acid , an inhibitor of N'-methyl-D-aspartate receptors , abolishes glutamatergic activation of P27361 REA / 2 but not the activation of P42345 REA - S6K ; the latter is completely abolished by inhibitors of voltage-dependent calcium channels . Added singly , dopamine gives slight , and norepinephrine a more significant , activation of P29323 REA and S6K ; both catecholeamines , however , enhance glutamatergic activation of S6K but not P29323 REA . After 12 days in culture , the response to direct glutamatergic activation is attenuated but can be uncovered by suppression of gamma-aminobutyric acid interneurons with bicuculline in the presence of the weak K ( + ) channel blocker 4 - aminopyridine ( DB06637 ) . This selective synaptic activation of P42345 REA - S6K is also resistant to APV and inhibited by Ca ( 2 + ) channel blockers and higher concentrations of glutamate . P13639 REA ( P13639 REA ) is phosphorylated and inhibited by the eEF 2 kinase ( P62158 kinase III ) ; the latter is inhibited by the S6K or Rsk . DB11562 / DB06637 or DB00761 - induced depolarization reduces , whereas higher concentrations of glutamate increases , P13639 REA phosphorylation . Thus the P42345 REA - S6K pathway in neurons , a critical component of the late phase of LTP , is activated by glutamatergic stimulation in a calcium / calmodulin-dependent fashion through a calcium pool controlled by postsynaptic voltage-dependent calcium channels , whereas sustained stimulation of extrasynaptic glutamate receptors is inhibitory .

Lycopene binding compromised PDGF-AA / - AB signaling and migration in smooth muscle cells and fibroblasts : prediction of the possible lycopene binding site within PDGF . Platelet-derived growth factor ( PDGF ) is a potent stimulator of growth and motility of smooth muscle cells ( SMCs ) and fibroblasts . Abnormalities of PDGF / PDGF receptor ( P09619 REA ) are thought to contribute to vascular diseases and malignancy . We previously showed that natural carotenoid lycopene can directly bind to DB00102 MEN and affect its related functions in vascular SMCs . In this study we examined lycopene effect on PDGF-AA / - AB-induced signaling and migration in SMCs and fibroblasts . We found that lycopene inhibited PDGF-AA-induced SMC and fibroblast migration in a concentration-dependent manner . Lycopene reduced PDGF-AA signaling , including phosphorylation in P16234 REA and its downstream protein kinases / enzymes . It also inhibited PDGF-AB-induced signaling and cell migration . However , lycopene did not affect PDGF-induced reactive oxygen species production and H2O2 - induced P09619 REA phosphorylation . The binding analysis revealed that lycopene but not beta-carotene could directly bind to PDGF-AA in vitro and in plasma and this binding competitively inhibited lycopene interaction with DB00102 MEN , suggesting that lycopene bound to PDGF-AA / - BB at a homologous / similar region within PDGF . Moreover , the docking and binding analyses predicted that the lycopene-binding region within PDGF was located at loop 2 region . Taken together , we provide here evidence that lycopene interacts with PDGF-AA / - AB and compromises their intracellular signaling , leading to a marked inhibition on PDGF-AA / - AB-induced migration in both SMCs and fibroblasts . We also predicted its binding region within PDGF and proved its anti-vascular injury effect in vivo . The results , together with our previous findings , suggest lycopene special affinity / effect for PDGF family and its possible application in prevention in vascular diseases and malignancy .

DB00502 MEN induces neurotoxicity by the DB01221 receptor downstream signaling pathway , alternative from glutamate excitotoxicity . The DB01221 receptor is believed to be important in a wide range of nervous system functions including neuronal migration , synapse formation , learning and memory . In addition , it is involved in excitotoxic neuronal cell death that occurs in a variety of acute and chronic neurological disorders . Besides of agonist / coagonist sites , other modulator sites , including butyrophenone site may regulate the N-methyl-D-aspartate receptor . It has been shown that haloperidol , an antipsychotic neuroleptic drug , interacts with the Q13224 REA subunit of DB01221 receptor and inhibits DB01221 response in neuronal cells . We found that DB01221 receptor was co-immunoprecipitated by anti-Ras antibody and this complex , beside NR2 subunit of DB01221 receptor contained haloperidol-binding proteins , P29475 REA and Ras - P01286 REA . Furthermore , we have shown that haloperidol induces neurotoxicity of neuronal cells via DB01221 receptor complex , accompanied by dissociation of Ras - P01286 REA from membranes and activation of c-Jun-kinase . Inclusion of insulin prevented relocalization of Ras - P01286 REA and subsequent neuronal death . DB00502 MENMAX DB00502 MEN - induced dissociation of Ras - P01286 REA leads to inhibition of membrane-bound form of Ras protein and changes downstream regulators activity that results in the initiation of the apoptotic processes via the mitochondrial way . Our results suggest that haloperidol induces neuronal cell death by the interaction with DB01221 receptor , but through the alternative from glutamate excitotoxicity signaling pathway .