interacts_with DB00126?
Prolyl-4-hydroxylase domain protein 2 controls NF-κB/p65 transactivation and enhances the catabolic effects of inflammatory cytokines on cells of the nucleus pulposus . Prolyl-4-hydroxylase ( P20941 ) proteins are key in sensing tissue hypoxia . In nucleus pulposus ( NP ) cells , our previous work demonstrated that P20941 isoforms have a differential contribution in controlling hypoxia-inducible factor ( HIF ) -α degradation and activity . Recently we have shown that a regulatory relationship exists between Q9H6Z9 and inflammatory cytokines in NP cells . With respect to Q9GZT9 , the most abundant P20941 isoform in NP cells , very little is known concerning its function and regulation under inflammatory conditions that characterize intervertebral disc degeneration . Here , we show that Q9GZT9 is a potent regulator of the catabolic activities of P01375 -α ; silencing of Q9GZT9 significantly decreased P01375 -α-induced expression of catabolic markers including P31431 , P08254 , P45452 , and Q9UNA0 , as well as several inflammatory cytokines and chemokines , while partially restoring aggrecan and collagen II expression . Use of NF-κB reporters with ShPHD2 , SiHIF-1α , as well as p65(-/-) , Q9GZT9 (-/-) , and Q9H6Z9 (-/-) cells , shows that Q9GZT9 serves as a co-activator of NF-κB/p65 signaling in Q9BYW2 -independent fashion . Immunoprecipitation of endogenous and exogenously expressed tagged proteins , as well as fluorescence microscopy , indicates that following P01375 -α treatment , Q9GZT9 interacts and co-localizes with p65 . Conversely , loss of function experiments using lentivirally delivered Sh-p65 , Sh-IKKβ , and NF-κB inhibitor confirmed that cytokine-dependent Q9GZT9 expression in NP cells requires NF-κB signaling . These findings clearly demonstrate that Q9GZT9 forms a regulatory circuit with P01375 -α via NF-κB and thereby plays an important role in enhancing activity of this cytokine . We propose that during disc degeneration Q9GZT9 may offer a therapeutic target to mitigate the deleterious actions of P01375 -α , a key proinflammatory cytokine . A P04035 inhibitor possesses a potent anti-atherosclerotic effect other than serum lipid lowering effects -- the relevance of endothelial nitric oxide synthase and superoxide anion scavenging action . We have determined whether the anti-atherosclerotic effect of a 3-hydroxy-3-methyl-glutaryl- DB01992 ( HMG- DB01992 ) reductase inhibitor ( fluvastatin ) is mediated through nitric oxide ( NO ) as well as affecting plasma lipids . NO related vascular responses , endothelial nitric oxide synthase ( P29474 ) mRNA and superoxide anion ( O(2)(-) ) release were examined in vascular walls of oophorectomized female rabbits fed 0.5 % cholesterol chow for 12 weeks with or without fluvastatin ( 2 mg/kg per day ) . Serum lipid profile was not different between two groups . NO dependent responses stimulated by acetylcholine and calcium ionophore A23187 and tone related basal NO response induced by N(G)-monomethyl-L-arginine acetate ( L- Q13145 ) ; nitric oxide synthase inhibitor were all improved by fluvastatin treatment . Endothelium independent vasorelaxation induced by nitroglycerin was not different between the two groups of rabbits ' arteries . DB01095 treatment increased cyclic GMP concentration in aorta of rabbits . P29474 mRNA expression and O(2)(-) release were measured in aorta using competitive reverse transcription-polymerase chain reaction ( RT-PCR ) and with lucigenin analogue , 2-methyl-3,7-dihydroimidazol [1,2-a]pyrazine-3-one ( MCLA ) chemiluminescence methods . P29474 mRNA in the endothelial cells of aorta was significantly up-regulated and O(2)(-) production was significantly reduced in fluvastatin treated rabbit aorta . Anti-macrophage staining area , but not anti-smooth muscle cell derived actin stained area in the aorta was also reduced by fluvastatin treatment . Conclusion , fluvastatin , a P04035 inhibitor , retards the initiation of atherosclerosis formation through the improvement of NO bioavailability by both up-regulation of P29474 mRNA and decrease of O(2)(-) production in vascular endothelial cells , and this means that part of the anti-atherosclerotic effect of fluvastatin may be due to nonlipid factors . P01138 deprivation-induced gene expression : after ten years , where do we stand ? Nerve growth factor ( P01138 ) is required for the survival of developing sympathetic and sensory neurons . In the absence of P01138 , these neurons undergo protein synthesis-dependent apoptosis . Ten years have gone by since the first reports of specific genes being upregulated during P01138 deprivation-induced cell death . Over the last decade , a few additional genes ( DP5 , Bim , Q9GZT9 ) have been added to a list that began with cyclin D1 and c-jun . In this chapter , we discuss the evidence that these genes act as regulators of neuronal cell death . We also suggest a hypothesis for how one gene , Q9GZT9 , may function to suppress a self-protection mechanism in P01138 -deprived neurons . Adulthood nicotine treatment alleviates behavioural impairments in rats neonatally treated with quinpirole : possible roles of acetylcholine function and neurotrophic factor expression . Increases in dopamine D(2) receptor sensitivity are known to be common in drug abuse and neurological disorders . Past data from this laboratory have shown that long-term increases in D(2) sensitivity can be produced by quinpirole treatment ( a D(2)/D(3) agonist ) during early development . The present investigation was designed to test the hypothesis that nicotine administration in adulthood would reduce both cognitive and skilled reaching impairments produced by increases in D(2) sensitivity . Female Sprague-Dawley rats were treated with quinpirole ( 1 mg/kg ) or saline from postnatal day 1 ( PD 1 ) to PD 21 . Beginning in adulthood ( PD 61 ) , rats were treated with nicotine ( 0.3 mg/kg free base ) or saline twice daily for 14 consecutive days before behavioural testing commenced . Animals neonatally treated with quinpirole demonstrated performance deficits on the Morris water task and a skilled reaching task compared to controls . Deficits on both tasks were completely alleviated by adulthood nicotine treatment . Animals neonatally treated with quinpirole demonstrated a significant 36 % decrease of P28329 in the hippocampus compared to saline controls that was partially eliminated by nicotine . Additionally , neonatal quinpirole produced a significant decrease in hippocampal P01138 content compared to controls , however , nicotine failed to alleviate this decrease in P01138 . The results of this investigation demonstrate that long-term increases in dopamine D(2) receptor sensitivity produce significant decreases in hippocampal cholinergic and P01138 expression that may result in cognitive impairment . DB00184 alleviates both cognitive and skilled reaching impairments caused by increases in D(2) sensitivity , but the mechanism through which nicotine is acting is currently unknown . 17 DB00783 -mediated growth inhibition of MDA-MB-468 cells stably transfected with the estrogen receptor : cell cycle effects . P03372 ( ER ) -negative MDA-MB-468 human breast cancer cells were stably transfected with wild-type human ER and utilized as a model for investigating estrogen- and aryl hydrocarbon ( Ah ) -responsiveness . Treatment of the stably transfected cells with 10 nM 17 beta-estradiol ( E2 ) resulted in a significant inhibition ( > 60 % ) of cell proliferation and DNA synthesis , which was blocked by 10(-7) M ICI 182 780 . Analysis by flow cytometry indicated that treatment with E2 increased the percentage of cells in G0/ P55008 ( from 68.8 to 89.4 ) and decreased cells in S ( from 18.4 to 3.4 ) and G2/M ( from 12.8 to 7.2 ) phases of the cell cycle . The effects of E2 on the major cyclins , cyclin-dependent kinases and cyclin-dependent kinase inhibitors , retinoblastoma protein ( RB ) , Q01094 , and cyclin-dependent kinase activities were also investigated in the stably transfected MDA-MB-468 cells . The results demonstrated that the growth inhibitory effects of 10(-8) M E2 in ER stably transfected MDA-MB-468 cells were associated with modulation of several factors required for cell cycle progression and DNA synthesis , including significant induction of the cyclin-dependent kinase inhibitor p21cip-1 ( > 4-fold increase after 12 h ) and decreased Q01094 and P12004 protein levels . These results show that the growth-inhibitory effects of E2 in the stably transfected cells were due to multiple factors which result in growth arrest in G0/ P55008 and inhibition of DNA synthesis . Serotonergic mechanisms in human allergic contact dermatitis . Expression of serotonin ( 5-hydroxytryptamine ; 5-HT ) , 5-HT receptors 1A ( 5-HT1AR ) and 2A , and serotonin transporter protein ( P31645 ) was studied in positive epicutaneous reactions to nickel sulphate in nickel-allergic patients , at 72 h post-challenge with the antigen . In addition , the effects of 5-HT2AR agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , and the selective serotonin reuptake inhibitors ( SSRIs ) citalopram and fluoxetine , were tested on nickel-stimulated peripheral blood mononuclear cells from nickel-allergic patients , regarding their proliferation and interleukin ( IL ) -2 production , as well as the effect of these SSRIs on a murine Langerhans ' cell-like line ( XS52 ) , regarding its IL-1beta production . Serotonin-positive platelets were increased in the inflamed skin compared with control skin . A decrease ( p < 0.01 ) in 5-HT1AR-positive mononuclear cells was evident in the eczematous skin compared with control skin , whereas 5-HT2AR- and P31645 -positive cells were increased ( p < 0.001 for both ) in the eczematous skin . Treatment of nickel-stimulated peripheral blood mononuclear cells with 5x10(-5) mol/l of DOI inhibited ( p < 0.01 ) the proliferation of nickel-stimulated peripheral blood mononuclear cells , while no effect was found regarding P60568 production . DB00215 at 10(-6) mol/l tended to inhibit the production of IL-1beta by the XS52 cell line . These results indicate the implication of the serotonergic system in the contact allergic reaction . Candidate gene studies of ADHD : a meta-analytic review . Quantitative genetic studies ( i.e. , twin and adoption studies ) suggest that genetic influences contribute substantially to the development of attention deficit hyperactivity disorder ( ADHD ) . Over the past 15 years , considerable efforts have been made to identify genes involved in the etiology of this disorder resulting in a large and often conflicting literature of candidate gene associations for ADHD . The first aim of the present study was to conduct a comprehensive meta-analytic review of this literature to determine which candidate genes show consistent evidence of association with childhood ADHD across studies . The second aim was to test for heterogeneity across studies in the effect sizes for each candidate gene as its presence might suggest moderating variables that could explain inconsistent results . Significant associations were identified for several candidate genes including Q01959 , P21917 , P21918 , P31645 , P28222 , and P60880 . Further , significant heterogeneity was observed for the associations between ADHD and Q01959 , P21917 , P21918 , P09172 , P08913 , P31645 , Q8IWU9 , P21397 , and P60880 , suggesting that future studies should explore potential moderators of these associations ( e.g. , ADHD subtype diagnoses , gender , exposure to environmental risk factors ) . We conclude with a discussion of these findings in relation to emerging themes relevant to future studies of the genetics of ADHD . A lactate-induced response to hypoxia . Organisms must be able to respond to low oxygen in a number of homeostatic and pathological contexts . Regulation of hypoxic responses via the hypoxia-inducible factor ( HIF ) is well established , but evidence indicates that other , HIF-independent mechanisms are also involved . Here , we report a hypoxic response that depends on the accumulation of lactate , a metabolite whose production increases in hypoxic conditions . We find that the Q9UGV2 protein is degraded in a Q9GZT9 / P40337 -dependent manner in normoxia but is protected from destruction by binding to lactate that accumulates under hypoxia . The stabilized Q9UGV2 protein binds c-Raf to mediate hypoxia-induced activation of Raf- P29323 pathway , promoting angiogenesis and cell growth . Inhibiting cellular lactate production abolishes the Q9UGV2 -mediated hypoxia responses . Our study , therefore , elucidates the molecular basis for lactate-induced hypoxia signaling , which can be exploited for the development of therapies targeting hypoxia-induced diseases . Hypoxia-inducible factor 1 is activated by dysregulated cyclin E during mammary epithelial morphogenesis . Increased cyclin E expression has been identified in human tumors of diverse histologies , and in studies of primary breast cancers , high cyclin E is associated with poor prognosis . We have studied dysregulated cyclin E in epithelial tissues using organotypic cultures of human mammary epithelial cells and a murine model . We unexpectedly discovered that dysregulated cyclin E impairs normal acinar morphogenesis in vitro , and this is associated with the induction of P38936 (Cip1) , p27(Kip1) , and cellular senescence . P12004 E-induced morphogenesis arrest is dependent upon hypoxia-inducible factor 1α ( HIF-1α ) , which itself is induced by high cyclin E both in cultured mammary acini and in mammary epithelial tissues in a mouse model of deregulated cyclin E expression . We next determined that E2F activity directly regulates and is required for induction of Q16665 by cyclin E. Additionally , we found that cyclin E deregulation in mammary acini decreases , in an E2F-independent manner , expression of the Q9GZT9 prolyl hydroxylase that regulates HIF-1α degradation within the P40337 ubiquitin ligase pathway . Together , our findings reveal a direct link between cyclin E and Q9BYW2 activities in mammary epithelial cells and implicate Q9BYW2 as a mediator of proliferation-independent phenotypes associated with high cyclin E expression in some human breast cancers . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . DB01037 transdermal system : in the treatment of major depressive disorder . The monamine oxidase ( MAO ) inhibitor selegiline is selective for P27338 at the low oral dosages used in the treatment of Parkinson 's disease . However , P21397 is also inhibited at the high oral dosages needed to effectively treat depression ( not an approved indication ) , necessitating a tyramine-restricted diet . The selegiline transdermal system was designed to deliver antidepressant drug concentrations to the CNS , without substantially impairing small intestine P21397 activity . At the target dose of 6 mg/24 hours , tyramine dietary restrictions are not needed . Short-term treatment with fixed ( 6 mg/24 hours ) or flexible ( 6 , 9 or 12 mg/24 hours ) doses of selegiline transdermal system was superior to placebo on most measures of antidepressant activity in 6- or 8-week , randomised , double-blind , multicentre studies in adult outpatients with major depressive disorder ( MDD ) . Likewise , long-term treatment with a fixed dose of selegiline transdermal system 6 mg/24 hours was superior to placebo as maintenance therapy in a 52-week , randomised , double-blind , multicentre , relapse-prevention trial in patients with MDD . DB01037 transdermal system therapy was generally well tolerated in placebo-controlled studies ; application site reactions , mostly of mild to moderate severity , were the most commonly reported adverse events . The incidence of sexual adverse effects and weight gain was low and similar to that with placebo . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . The low-potency , voltage-dependent Q12809 blocker propafenone -- molecular determinants and drug trapping . The molecular determinants of high-affinity human ether-a-go-go-related gene ( Q12809 ) potassium channel blockade by methanesulfonanilides include two aromatic residues ( Phe656 and Tyr652 ) on the inner helices ( S6 ) and residues on the pore helices that face into the inner cavity , but determinants for lower-affinity Q12809 blockers may be different . In this study , alanine-substituted Q12809 channel mutants of inner cavity residues were expressed in Xenopus laevis oocytes and were used to characterize the Q12809 channel binding site of the antiarrhythmic propafenone . DB01182 's blockade of Q12809 was strongly dependent on residue Phe656 but was insensitive or weakly sensitive to mutation of Tyr652 , Thr623 , Ser624 , Val625 , Gly648 , or Val659 and did not require functional inactivation . Homology models of Q12809 based on KcsA and MthK crystal structures , representing the closed and open forms of the channel , respectively , suggest propafenone is trapped in the inner cavity and is unable to interact exclusively with Phe656 in the closed state ( whereas exclusive interactions between propafenone and Phe656 are found in the open-channel model ) . These findings are supported by very slow recovery of wild-type Q12809 channels from block at -120 mV , but extremely rapid recovery of D540K channels that reopen at this potential . The experiments and modeling suggest that the open-state propafenone binding-site may be formed by the Phe656 residues alone . The binding site for propafenone ( which may involve pi-stacking interactions with two or more Phe656 side-chains ) is either perturbed or becomes less accessible because of closed-channel gating . This provides further evidence for the existence of gating-induced changes in the spatial location of Phe656 side chains . Altered gene expression by low-dose arsenic exposure in humans and cultured cardiomyocytes : assessment by real-time PCR arrays . Chronic arsenic exposure results in higher risk of skin , lung , and bladder cancer , as well as cardiovascular disease and diabetes . The purpose of this study was to investigate the effects on expression of selected genes in the blood lymphocytes from 159 people exposed chronically to arsenic in their drinking water using a novel RT-PCR TaqMan low-density array ( TLDA ) . We found that expression of tumor necrosis factor-α ( P01375 -α ) , which activates both inflammation and NF-κB-dependent survival pathways , was strongly associated with water and urinary arsenic levels . Expression of P22460 , which encodes a potassium ion channel protein , was positively associated with water and toe nail arsenic levels . Expression of 2 and 11 genes were positively associated with nail and urinary arsenic , respectively . Because arsenic exposure has been reported to be associated with long QT intervals and vascular disease in humans , we also used this TLDA for analysis of gene expression in human cardiomyocytes exposed to arsenic in vitro . Expression of the ion-channel genes CACNA1 , Q12809 , P51787 and P15382 were down-regulated by 1-μM arsenic . Alteration of some common pathways , including those involved in oxidative stress , inflammatory signaling , and ion-channel function , may underlay the seemingly disparate array of arsenic-associated diseases , such as cancer , cardiovascular disease , and diabetes . Aerosol vaccination with AERAS-402 elicits robust cellular immune responses in the lungs of rhesus macaques but fails to protect against high-dose Mycobacterium tuberculosis challenge . Development of a vaccine against pulmonary tuberculosis may require immunization strategies that induce a high frequency of Ag-specific P01730 and CD8 T cells in the lung . The nonhuman primate model is essential for testing such approaches because it has predictive value for how vaccines elicit responses in humans . In this study , we used an aerosol vaccination strategy to administer AERAS-402 , a replication-defective recombinant adenovirus ( rAd ) type 35 expressing Mycobacterium tuberculosis Ags Ag85A , Ag85B , and TB10.4 , in bacillus Calmette-Guérin ( BCG ) -primed or unprimed rhesus macaques . Immunization with BCG generated low purified protein derivative-specific P01730 T cell responses in blood and bronchoalveolar lavage . In contrast , aerosolized AERAS-402 alone or following BCG induced potent and stable Ag85A/b-specific P01730 and CD8 effector T cells in bronchoalveolar lavage that largely produced IFN-γ , as well as P01375 and P60568 . Such responses induced by BCG , AERAS-402 , or both failed to confer overall protection following challenge with 275 CFUs M. tuberculosis Erdman , although vaccine-induced responses associated with reduced pathology were observed in some animals . Anamnestic T cell responses to Ag85A/b were not detected in blood of immunized animals after challenge . Overall , our data suggest that a high M. tuberculosis challenge dose may be a critical factor in limiting vaccine efficacy in this model . However , the ability of aerosol rAd immunization to generate potent cellular immunity in the lung suggests that using different or more immunogens , alternative rAd serotypes with enhanced immunogenicity , and a physiological challenge dose may achieve protection against M. tuberculosis . DB00126 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 Cys201Ser 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 . Disparate effects of simvastatin on angiogenesis during hypoxia and inflammation . AIMS : Studies have shown that some of statin 's pleiotropic effects were achieved by either promotion or inhibition of angiogenesis , depending on the underlying disease . This study tested the hypothesis that the angiogenic potential of simvastatin is related to the microenvironmental conditions . MAIN METHODS : Human umbilical vein endothelial cells ( HUVEC ) were studied after exposure to hypoxia or the inflammatory factors tumor necrosis factor ( P01375 ) -alpha , with or without co-incubation with simvastatin ( 1 micromol/L ) and mevalonate . HUVEC angiogenesis was evaluated by tube formation , migration , and proliferation assays . Hypoxia inducible factor ( HIF ) -1alpha , vascular endothelial growth factor ( P15692 ) , Akt , endothelium nitric oxide synthase ( e-NOS ) , and oxidative stress were evaluated . KEY FINDINGS : HUVEC angiogenesis increased during hypoxia ( tube length 14.7+/-0.5 vs. 7.8+/-0.6 mm , p < 0.05 ) and further enhanced by simvastatin ( 19.3+/-1.1 mm , p < 0.05 vs. hypoxia alone ) , which downregulated the expression of the Q9BYW2 inhibitor Q9GZT9 and upregulated HIF-1alpha , P15692 , and Akt , without changing oxidative stress or P29474 . Incubation with P01375 promoted HUVEC angiogenesis ( 7.4+/-0.2 vs. 6.5+/-0.2 mm , p < 0.05 ) with increased oxidative stress . However , simvastatin inhibited this promotion ( 2.5+/-0.3 mm , p < 0.001 vs. P01375 alone ) by decreasing oxidative stress , P15692 , Akt , and P29474 . SIGNIFICANCE : We conclude that at the same dosage , simvastatin can either promote or inhibit angiogenesis , possibly by activating upstream regulators of HIF-1alpha in hypoxia , but conversely interfering with angiogenic signaling downstream to inflammation . These opposing angiogenic effects should be considered in the therapeutic strategies with statins . Suppression of NF-kappaB activity by sulfasalazine is mediated by direct inhibition of IkappaB kinases alpha and beta . BACKGROUND & AIMS : Activation of NF-kappaB/Rel has been implicated in the pathogenesis of inflammatory bowel disease ( Q9UKU7 ) . Various drugs used in the treatment of Q9UKU7 , such as glucocorticoids , DB00244 , and sulfasalazine , interfere with NF-kappaB/Rel signaling . The aim of this study was to define the molecular mechanism by which sulfasalazine inhibits NF-kappaB activation . METHODS : The effects of sulfasalazine and its moieties on NF-kappaB signaling were evaluated using electromobility shift , transfection , and immune complex kinase assays . The direct effect of sulfasalazine on O15111 ( IKK ) activity was investigated using purified recombinant O15111 and -beta proteins . RESULTS : NF-kappaB/Rel activity induced by tumor necrosis factor alpha , 12-O-tetradecanoylphorbol-13-acetate , or overexpression of NF-kappaB-inducing kinase , O15111 , O14920 , or constitutively active O15111 and O14920 mutants was inhibited dose dependently by sulfasalazine . Sulfasalazine inhibited tumor necrosis factor alpha-induced activation of endogenous IKK in Jurkat T cells and SW620 colon cells , as well as the catalytic activity of purified O15111 and O14920 in vitro . In contrast , the moieties of sulfasalazine , DB00244 , and sulfapyridine or DB00233 had no effect . Activation of extracellular signal-related kinase ( P29323 ) 1 and 2 , c-Jun-N-terminal kinase ( JNK ) 1 , and p38 was unaffected by sulfasalazine . The decrease in substrate phosphorylation by O15111 and -beta is associated with a decrease in autophosphorylation of IKKs and can be antagonized by excess adenosine triphosphate . CONCLUSIONS : These data identify sulfasalazine as a direct inhibitor of O15111 and -beta by antagonizing adenosine triphosphate binding . The suppression of NF-kappaB activation by inhibition of the IKKs contributes to the well-known anti-inflammatory and immunosuppressive effects of sulfasalazine . DB00435 donor , (+/-)-S-nitroso-N-acetylpenicillamine , stabilizes transactive hypoxia-inducible factor-1alpha by inhibiting von Hippel-Lindau recruitment and asparagine hydroxylation . We have confirmed that the NO donor (+/-)-S-nitroso-N-acetylpenicillamine ( P60880 ) stabilizes the transactive form of hypoxia-inducible factor-1alpha ( HIF-1alpha ) , leading to the induction of HIF-1alpha target genes such as vascular endothelial growth factor and carbonic anhydrase 9 . Activation of HIF-1alpha should require inhibition of the dual system that keeps it inactive . One is ubiquitination , which is triggered by hydroxylation of HIF-1alpha-proline and the subsequent binding of E3 ubiquitin ligase , the von Hippel Lindau ( P40337 ) protein . The other is hydroxylation of HIF-1alpha-asparagine , which reduces the affinity of HIF-1alpha for its coactivator , DB02527 responsive element binding protein/p300 . We examined the effects of the NO donor P60880 on proline and asparagine hydroxylation of HIF-1alpha peptides by measuring the activities of the corresponding enzymes , HIF-1alpha-specific proline hydroxylase 2 ( Q9GZT9 ) and the HIF-1alpha-specific asparagine hydroxylase , designated factor inhibiting HIF-1alpha ( Q9NWT6 ) , respectively . We found that the P60880 did not prevent Q9GZT9 from hydroxylating the proline of HIF-1alpha . Instead , it blocked the interaction between P40337 and the proline-hydroxylated HIF-1alpha , but only when the reducing agents Fe(II) and vitamin C were limiting . The fact that the absence of cysteine 520 of HIF-1alpha abolishes its responsiveness to P60880 suggests that this residue mediates the inhibition by P60880 of the interaction between P40337 and HIF-1alpha , presumably by S-nitrosylation of HIF-1alpha . Un-like Q9GZT9 , asparagine hydroxylation by Q9NWT6 was directly inhibited by P60880 , but again only when reducing agents were limiting . Substitution of cysteine 800 of HIF-1alpha with alanine failed to reverse the inhibitory effects of P60880 on asparagine hydroxylation , implying that Q9NWT6 , not its substrate HIF-1alpha , is inhibited by P60880 . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . A study of 36 unrelated cases with pure erythrocytosis revealed three new mutations in the erythropoietin receptor gene . Thirty-six unrelated cases with erythrocytosis of unknown origin were investigated . Exons 5-8 of the erythropoietin receptor gene ( P19235 ) , the von Hippel-Lindau gene , and the prolyl hydroxylase domain protein 2 gene ( Q9GZT9 ) were screened by direct DNA sequencing . The O60674 mutation , O60674 ( Val617Phe ) , was screened by allele specific PCR . In this study , three new mutations of P19235 causing deletions in exon 8 were found : the first led directly to a stop codon [ g.5957_5958delTT ( p.Phe424X ) ] , the second to a stop codon after one residue [ g.5828_5829delCC ( p.Pro381GlnfsX1 ) ] and the third to a stop codon following a frameshift sequence of 23 residues [ g.5971delC ( p.Leu429TrpfsX23 ) ] . One patient had a previously reported P19235 mutation [ g.6146A > G ( p.Asn487Ser ) ] and another , a silent one ( g.5799G > A ) . All were heterozygotes . In addition , 2 patients were positive for O60674 ( Val617Phe ) , and 2 reported elsewhere , were mutated in the Q9GZT9 gene [ c.606delG ( p.Met202IlefsX71 ) .