MH_dev_273

Secretagogue-stimulated pancreatic secretion is differentially regulated by constitutive NOS isoforms in mice . DB00435 ( NO ) and NO synthase ( NOS ) play controversial roles in pancreatic secretion . NOS inhibition reduces CCK-stimulated in vivo pancreatic secretion , but it is unclear which NOS isoform is responsible , because NOS inhibitors lack specificity and three NOS isoforms exist : neuronal ( P29475 REA ) , endothelial ( P29474 REA ) , and inducible ( P35228 REA ) . Mice having individual NOS gene deletions were used to clarify the NOS species and cellular interactions influencing pancreatic secretion . In vivo secretion was performed in anesthetized mice by collecting extraduodenal pancreatic duct juice and measuring protein output . Nonselective NOS blockade was induced with N ( omega ) - nitro-L-arginine ( DB04223 MEN ; 10 mg / kg ) . In vivo pancreatic secretion was maximal at 160 pmol.kg ( - 1 ) . h ( - 1 ) CCK octapeptide ( CCK - 8 ) and was reduced by NOS blockade ( 45 % ) and P29474 REA deletion ( 44 % ) . Secretion was unaffected by P35228 REA deletion but was increased by P29475 REA deletion ( 91 % ) . To determine whether the influence of NOS on secretion involved nonacinar events , in vitro CCK - 8 - stimulated secretion of amylase from isolated acini was studied and found to be unaltered by NOS blockade and P29474 REA deletion . Influence of NOS on in vivo secretion was further examined with carbachol . Protein secretion , which was maximal at 100 nmol.kg ( - 1 ) . h ( - 1 ) carbachol , was reduced by NOS blockade and P29474 REA deletion but unaffected by P29475 REA deletion . NOS blockade by DB04223 MEN had no effect on carbachol-stimulated amylase secretion in vitro . Thus constitutive NOS isoforms can exert opposite effects on in vivo pancreatic secretion . P29474 REA likely plays a dominant role , because P29474 REA deletion mimics NOS blockade by inhibiting CCK - 8 and carbachol-stimulated secretion , whereas P29475 REA deletion augments CCK - 8 but not carbachol-stimulated secretion .

P11511 REA inhibitors and cyclooxygenase - 2 ( P35354 REA ) inhibitors in endometriosis : new questions - - old answers ? The medical treatment of endometriosis needs to be optimized . Therapeutic management strategies for endometriosis-associated pain or recurrent disease are primarily aimed at downregulating ovarian function or antagonizing the effect of estrogen in ectopic endometrial implants . In this context , basic research is providing important results for the development of new , specific treatment modalities . P11511 REA overexpression has recently been detected in endometriotic tissue . P11511 REA ( p450arom ) is responsible for converting C19 androgens into estrogen in several types of human tissue . P11511 REA activity causes local estrogen biosynthesis , which , in turn , stimulates prostaglandin E2 production by upregulating cyclooxygenase - 2 ( P35354 REA ) . Thus , a positive feedback cycle develops between the two systems . Another abnormality in endometriosis , the deficient 17beta - hydroxysteroiddehydrogenase type II ( 17beta - HSD-Type-II ) expression , impairs the inactivation of estradiol to estrone . In contrast to the eutopic endometrium , these molecular aberrations increase the amount of local estradiol and prostaglandin E2 in endometriosis . In several human cell lines , prostaglandin and estrogen concentrations are associated with proliferation , migration , angiogenesis , apoptosis resistance and even invasiveness . Consequently , aromatase and P35354 REA are thought to be promising new therapeutic targets . Thus , specific aromatase inhibitors ( e . g . DB01006 MEN / DB01006 MEN , DB01217 / Arimidex or Exemestan / Aromasin ) or selective P35354 REA inhibitors ( e . g . Celecoxib / DB00482 , DB00533 / Vioxx , DB00580 / Bextra ) are of great interest and should be studied in clinical trials in premenopausal woman with endometriosis to expand the spectrum of currently available treatment options .

Inhibitors of the interaction between P04275 REA and platelet GPIb / IX / V . The formation of platelet-rich thrombi , a critical step in the pathogenesis of atherothrombotic events , is a multistep process involving several components , among which von Willebrand Factor ( P04275 REA ) plays a central role . Ruptured atherosclerotic plaques expose subendothelial matrix proteins which bind P04275 REA that represents a bridge between the injured blood vessel and activated platelets , playing a crucial role in platelet adhesion and aggregation , especially in conditions of high-shear rate . Due to these peculiarities , the binding of P04275 REA to GPIbα is an attractive drug target . Here we summarize the present knowledge on the different classes of drugs targeting the P04275 REA - GPIb interaction and we give an account of their level of clinical development . In particular , the following compounds are discussed : AJW 200 , an IgG 4 humanized monoclonal antibody against P04275 REA - A1 ; 82D6A3 , a monoclonal antibody against P04275 REA - A3 ; Q96JZ2 - 0081 and Q96JZ2 - 0681 , bivalent humanized nanobodies targeting the P04275 REA - A1 domain ; DB05202 MEN and its advanced formulation ARC 15105 , second-generation aptamers that bind the P04275 REA - A1 domain ; h6B4 - Fab , a murine monoclonal antibody , and GPG - 290 , a recombinant chimeric protein , both directed against GPIbα .

DB00107 MEN and vasopressin receptor polymorphisms interact with circulating neuropeptides to predict human emotional reactions to stress . DB00107 MEN ( OT ) and a polymorphism ( rs53576 ) in the oxytocin receptor gene ( P30559 REA ) have been independently associated with stress reactivity , whereas oxytocin ' s sister peptide , arginine vasopressin ( AVP ) and polymorphisms in the vasopressin receptor gene ( P37288 REA ) have been independently associated with aggressive behavior . In this study , 68 men and 98 women were genotyped for the P30559 REA rs53576 polymorphism and the P37288 REA O15537 polymorphism . Baseline and poststressor levels of plasma OT , plasma AVP , positive affect , and anger were assessed . Women , but not men , with high levels of poststressor OT and the GG genotype of rs53576 felt the most positive affect after the stressor . Men , but not women , with high levels of poststressor AVP and the 320 allele of the O15537 polymorphism reported more poststressor anger than noncarriers . These data constitute the first evidence that oxytocin and vasopressin receptor genes interact with levels of OT and AVP to predict sex-specific emotional stress responses .

A pleiotropic antiatherogenic action of ibuprofen . Ibuprofen is a cyclooxygenase ( P23219 REA and P35354 REA ) inhibitor known to reduce the production of prostaglandins that play prominent role in inflammation . Other properties of the drug , aside from its anti-inflammatory effects , have been recently studied . In this paper we shall discuss several properties of ibuprofen that making the drug interesting for treatment of conditions associated with atherosclerosis . Ibuprofen exerts pleiotropic effects such as inhibition of adhesion and transendothelial migration of leukocytes , suppressing intracellular production of reactive oxygen species and oxidative modification of LDL . Interestingly , ibuprofen increased HDL cholesterol levels and reduced the level of triglicerides . Ibuprofen can also modulate efficiency of fibrynolisis by inhibiting production of plasminogen activator inhibitor ( P05121 REA ) . This properties of ibuprofen may be due to changing the activity of transcription factors . Ibuprofen inhibits the activation of NF-kB and activates PPARa and PPARg .

Direct anti-inflammatory mechanisms contribute to attenuation of experimental allograft arteriosclerosis by statins . BACKGROUND : Despite the development of effective immunosuppressive therapy , transplant graft arterial disease ( Q99259 REA ) remains the major limitation to long-term graft survival . The interplay between host inflammatory cells and donor vascular wall cells results in an intimal hyperplastic lesion , which leads to ischemia and graft failure . P04035 REA inhibitors ( statins ) reduce Q99259 REA in human cardiac allografts , although it is unclear whether this is secondary to cholesterol lowering or other mechanisms . This study tested the hypothesis that statins can suppress Q99259 REA by cholesterol-independent pathways . METHODS AND RESULTS : We performed heterotopic murine cardiac transplants in total allogeneic or major histocompatibility complex class II-mismatched combinations . Transplanted animals received either control chow , chow containing 25 ppm cerivastatin ( low dose ) , or chow containing 125 ppm cerivastatin ( high dose ) . Mean plasma cerivastatin concentrations were 0.0 ( control ) , 10.1 ( low dose ) , and 21.9 ( high dose ) nmol / L , respectively . Plasma cholesterol levels were the same in all groups . Q99259 REA scores decreased in low-dose ( P < 0.05 ) and high-dose ( P < 0.0001 ) cerivastatin groups compared with controls , with concomitant reduction in graft-infiltrating cells and significantly decreased intragraft RANTES and monocyte chemotactic protein - 1 mRNA expression . DB00439 MEN , as well as other statins , also reduced RANTES and monocyte chemotactic protein - 1 production in mouse endothelial cells stimulated with interferon-gamma and tumor necrosis factor-alpha in vitro . CONCLUSIONS : Clinically achievable levels of an P04035 REA inhibitor attenuate Q99259 REA in murine heart transplants , diminish host inflammatory cell recruitment , and do not alter cholesterol levels . These results indicate that statins can affect arterial biology and inflammation independently of their effects on cholesterol metabolism .

Acute renal failure from hemoglobinuric and interstitial nephritis secondary to iodine and mefenamic acid . DB00784 MENMAX DB00784 MEN ingestion , usually in excess and over prolonged period is known to produce interstitial nephritis , or less commonly papillary necrosis , with acute renal failure . However , it is not dose-dependent for the induction of tubulointerstitial damage . Excess iodine ingestion is known to produce toxicity and possible death , but acute renal failure is rare . There is evidence from clinical and experimental data that iodine has toxic effect on tubular epithelial cells . Iodine has not been documented to produce red cell hemolysis and hemoglobinuria . We present a unique case of acute renal failure from hemoglobinuric and acute interstitial nephritis secondary to suicidal ingestion of potassium iodide solution and also ingestion of a few mefenamic acid tablets . These agents led to potentiation of the renal injury from hemoglobinuric tubulopathy , probably from the iodine , and renal dysfunction from alteration of renal perfusion by selective P23219 REA inhibition of prostaglandin production , and induction of acute interstitial nephritis from mefenamic acid , leading to acute renal failure which was reversible by hemodialysis and supportive therapy .

DB02010 MEN - induced growth inhibition of glioma cells is accompanied by altered expression of cyclins , CDKs and CDK inhibitors . DB02010 MEN was found to bring about complete growth inhibition of human glioma cell lines . U87 MG cells were arrested in S phase while U373 MG cells in G2 / M phase on staurosporine treatment . Consistent with this observation , no change in P55008 phase regulators viz . , P12004 REA D1 , D3 and P11802 REA was seen on staurosporine treatment . The levels of P24941 REA , P06493 REA , P12004 REA A and P12004 REA B proteins decreased , while the levels of CDK inhibitors viz . , P38936 REA and p27 were found to increase on staurosporine treatment . The mRNA levels of P24941 REA and P06493 REA genes were also found to decrease on staurosporine treatment . Thus apart from staurosporine ' s known direct inhibitory effect on P24941 REA and P06493 REA activities , staurosporine was found to down-regulate activities of these two kinases by modulating the expression of the kinases themselves as well that of their activating partners ( Cyclins ) and their inhibitors .

Comparison of low fat and low carbohydrate diets on circulating fatty acid composition and markers of inflammation . Abnormal distribution of plasma fatty acids and increased inflammation are prominent features of metabolic syndrome . We tested whether these components of metabolic syndrome , like dyslipidemia and glycemia , are responsive to carbohydrate restriction . Overweight men and women with atherogenic dyslipidemia consumed ad libitum diets very low in carbohydrate ( VLCKD ) ( 1504 kcal : % CHO : fat :p rotein = 12:59 : 28 ) or low in fat ( LFD ) ( 1478 kcal : % CHO : fat :p rotein = 56:24 : 20 ) for 12 weeks . In comparison to the LFD , the VLCKD resulted in an increased proportion of serum total n - 6 PUFA , mainly attributed to a marked increase in arachidonate ( 20:4 n - 6 ) , while its biosynthetic metabolic intermediates were decreased . The n - 6 / n - 3 and arachidonic / eicosapentaenoic acid ratio also increased sharply . Total saturated fatty acids and 16:1 n - 7 were consistently decreased following the VLCKD . Both diets significantly decreased the concentration of several serum inflammatory markers , but there was an overall greater anti-inflammatory effect associated with the VLCKD , as evidenced by greater decreases in P01375 REA , P05231 REA , P10145 REA , P13500 REA , P16581 REA , I - P62158 , and P05121 REA . Increased 20:4 n - 6 and the ratios of 20:4 n -6/20 : 5n - 3 and n - 6 / n - 3 are commonly viewed as pro-inflammatory , but unexpectedly were consistently inversely associated with responses in inflammatory proteins . In summary , a very low carbohydrate diet resulted in profound alterations in fatty acid composition and reduced inflammation compared to a low fat diet .

DB00107 MEN attenuates NADPH-dependent superoxide activity and P05231 REA secretion in macrophages and vascular cells . DB00107 MEN is synthesized and released in the heart and vasculature , tissues that also express oxytocin receptors . Although it has been established this intrinsic cardiovascular oxytocin system is important in normal homeostatic cardiac and vascular regulation , a role for this system in cardiovascular pathophysiology has not been investigated . The current study examined the influence of oxytocin on mechanisms in atherogenesis , oxidative stress , and inflammation in cultured human vascular cells , THP - 1 monocytes , and macrophages . P30559 REA protein and mRNA expression , NADPH-dependent superoxide activity , and interleukin - 6 secretion were measured . Results demonstrated oxytocin receptor protein and mRNA in THP - 1 monocytes and macrophages . Incubation of cells at physiological levels of oxytocin significantly decreased basal and stimulated NADPH-dependent superoxide activity in vascular cells , monocytes , and macrophages by 24-48 % . DB00107 MEN also attenuated interleukin - 6 secretion from stimulated THP - 1 macrophages and endothelial cells by 56 and 26 % , respectively . These findings suggest that oxytocin attenuates vascular oxidative stress and inflammation , two important pathophysiological processes in atherosclerosis . The fact that oxytocin receptors are found in monocytes and macrophages , and oxytocin decreases both superoxide production and release of a proinflammatory cytokine from these cells , suggests a potentially larger role for oxytocin in the attenuation of disease .

BIO 5192 , a small molecule inhibitor of VLA - 4 , mobilizes hematopoietic stem and progenitor cells . Here we show that interruption of the P19320 REA / VLA - 4 axis with a small molecule inhibitor of VLA - 4 , BIO 5192 , results in a 30 - fold increase in mobilization of murine hematopoietic stem and progenitors ( HSPCs ) over basal levels . An additive affect on O14818 REA mobilization ( 3 - fold ) was observed when plerixafor ( DB06809 MEN ) , a small molecule inhibitor of the P61073 REA / P48061 REA axis , was combined with BIO 5192 . Furthermore , the combination of granulocyte colony-stimulating factor ( DB00099 ) , BIO 5192 , and plerixafor enhanced mobilization by 17 - fold compared with G - P04141 REA alone . HSPCs mobilized by BIO 5192 or the combination of BIO 5192 and plerixafor mobilized long-term repopulating cells , which successfully engraft and expand in a multilineage fashion in secondary transplantation recipients . Splenectomy resulted in a dramatic enhancement of G - P04141 REA - induced mobilization while decreasing both plerixafor - and BIO 5192 - induced mobilization of HSPCs . These data provide evidence for the utility of small molecule inhibitors of VLA - 4 either alone or in combination with G - P04141 REA or DB06809 MEN for mobilization of hematopoietic stem and progenitor cells .

Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 REA ( P05231 REA ) , C-Reactive Protein ( CRP ) , P00734 REA Fragments 1 and 2 ( F 1 + 2 ) , cortisol and P00747 REA Activator Inhibitor 1 ( P05121 REA ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 REA and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 REA level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge .

Targeting P04275 REA as a novel anti-platelet therapy ; application of DB05202 MEN , an Anti - P04275 REA aptamer , against thrombotic risk . Excessive activation of platelets is a causative factor for thrombotic diseases such as acute coronary syndrome or stroke , and various anti-platelet drugs were developed . DB00945 and clopidogrel have been used as gold standards for anti-platelet therapies , however , their clinical limitations including bleeding problem have increased the demand driving development of novel anti-platelet drugs with new targets . Among several activating pathways leading to platelet aggregation , the interaction between P04275 REA ( P04275 REA ) and glycoprotein Ib , which mainly occurs under high shear stress in arterioles , is recently suggested to be a new promising target . The anti-thrombotic efficacy of anti - P04275 REA agents , such as DB05202 MEN , has been proved in several preclinical and clinical studies . Here , we will discuss the potential benefits of targeting P04275 REA as a novel antiplatelet therapy , providing an insight into the role of P04275 REA in increased thrombotic risk .

DB01373 MEN - calmodulin suppresses the filamentous actin-binding activity of a 135 - kilodalton actin-bundling protein isolated from lily pollen tubes . We have isolated a 135 - kD actin-bundling protein ( P - 135 - P04278 ) from lily ( Lilium longiflorum ) pollen tubes and have shown that this protein is responsible for bundling actin filaments in lily pollen tubes ( E . Yokota , K . Takahara , T . Shimmen [ 1998 ] Plant Physiol 116 : 1421-1429 ) . However , only a few thin actin-filament bundles are present in random orientation in the tip region of pollen tubes , where high concentrations of Ca ( 2 + ) have also been found . To elucidate the molecular mechanism for the temporal and spatial regulation of actin-filament organization in the tip region of pollen tubes , we explored the possible presence of factors modulating the filamentous actin ( F-actin ) - binding activity of P - 135 - P04278 . The F-actin-binding activity of P - 135 - P04278 in vitro was appreciably reduced by Ca ( 2 + ) and calmodulin ( P62158 ) , although neither Ca ( 2 + ) alone nor P62158 in the presence of low concentrations of Ca ( 2 + ) affects the activity of P - 135 - P04278 . A micromolar order of Ca ( 2 + ) and P62158 were needed to induce the inhibition of the binding activity of P - 135 - P04278 to F-actin . An antagonist for P62158 , W - 7 , cancelled this inhibition . W - 5 also alleviated the inhibition effect of Ca ( 2 + ) - P62158 , however , more weakly than W - 7 . These results suggest the specific interaction of P - 135 - P04278 with Ca ( 2 + ) - P62158 . In the presence of both Ca ( 2 + ) and P62158 , P - 135 - P04278 organized F-actin into thin bundles , instead of the thick bundles observed in the absence of P62158 . These results suggest that the inhibition of the P - 135 - P04278 activity by Ca ( 2 + ) - P62158 is an important regulatory mechanism for organizing actin filaments in the tip region of lily pollen tubes .

Predictive value of circulating interleukin - 6 and heart-type fatty acid binding protein for three months clinical outcome in acute cerebral infarction : multiple blood markers profiling study . INTRODUCTION : There is no single blood marker for predicting the prognosis in ischemic stroke . A combination of multiple blood markers may enhance the ability to predict long-term outcome following ischemic stroke . METHODS : Blood concentrations of neuronal markers ( neuron-specific enolase , visinin-like protein 1 , heart type fatty acid binding protein ( hFABP ) and neuroglobin ) , astroglial markers ( P04271 REA and glial fibrillary acidic protein ) , inflammatory markers ( P05231 REA , P01375 REA - α , and P02741 REA ) , blood-brain barrier marker ( matrix metalloproteinase 9 ) , and haemostatic markers ( D-dimer and P05121 REA ) were measured within 24 hours after stroke onset . The discrimination and reclassification for favorable and poor outcome were compared after adding individual or a combination of blood markers to the clinical model of stroke outcome . RESULTS : In multivariate analysis , natural log-transformed ( log ) P05231 REA ( odds ratio ( OR ) : 1.75 , 95 % CI : 1.25 to 2.25 , P= 0.001 ) and loghFABP ( OR : 3.23 , 95 % CI : 1.44 to 7.27 , P= 0.005 ) were independently associated with poor outcome . The addition of a single blood marker to the clinical model did not improve the discriminating ability of the clinical model of stroke outcome . However , the addition of the combination of logIL - 6 and loghFABP to the clinical model showed improved discrimination ( area under receiver operating characteristic ( AUROC ) curve : 0.939 versus 0.910 , P= 0.03 ) and reclassification performance ( net reclassification improvement index : 0.18 , P= 0.005 ) . CONCLUSIONS : A combination of circulating P05231 REA and hFABP level has an additive clinical value for the prediction of stroke outcome .

Novel mechanism of endothelial nitric oxide synthase activation mediated by caveolae internalization in endothelial cells . Q03135 REA , the caveolae scaffolding protein , binds to and negatively regulates P29474 REA activity . As caveolin - 1 also regulates caveolae-mediated endocytosis after activation of the 60 - kDa albumin-binding glycoprotein gp60 in endothelial cells , we addressed the possibility that endothelial NO synthase ( P29474 REA ) - dependent NO production was functionally coupled to caveolae internalization . We observed that gp60 - induced activation of endocytosis increased NO production within 2 minutes and up to 20 minutes . NOS inhibitor N ( G ) - nitro-L-arginine ( DB04223 MEN ) prevented the NO production . To determine the role of caveolae internalization in the mechanism of NO production , we expressed dominant-negative dynamin - 2 mutant ( K44A ) or treated cells with methyl-beta-cyclodextrin . Both interventions inhibited caveolae-mediated endocytosis and NO generation induced by gp60 . We determined the role of signaling via Src kinase in the observed coupling of endocytosis to P29474 REA activation . Src activation induced the phosphorylation of caveolin - 1 , Akt and P29474 REA , and promoted dissociation of P29474 REA from caveolin - 1 . Inhibitors of Src kinase and Akt also prevented NO production . In isolated perfused mouse lungs , gp60 activation induced NO-dependent vasodilation , whereas the response was attenuated in P29474 REA ( - / - ) or caveolin - 1 ( - / - ) lungs . Together , these results demonstrate a critical role of caveolae-mediated endocytosis in regulating P29474 REA activation in endothelial cells and thereby the NO-dependent vasomotor tone .

P55008 arrest and down-regulation of cyclin E / cyclin-dependent kinase 2 by the protein kinase inhibitor staurosporine are dependent on the retinoblastoma protein in the bladder carcinoma cell line 5637 . The protein kinase inhibitor staurosporine has been shown to induce P55008 phase arrest in normal cells but not in most transformed cells . DB02010 MEN did not induce P55008 phase arrest in the bladder carcinoma cell line 5637 that lacks a functional retinoblastoma protein ( pRB - ) . However , when infected with a pRB-expressing retrovirus [ Goodrich , D . W . , Chen , Y . , Scully , P . & Lee , W . - H . ( 1992 ) Cancer Res . 52 , 1968-1973 ] , these cells , now pRB + , were arrested by staurosporine in P55008 phase . This arrest was accompanied by the accumulation of hypophosphorylated pRB . In both the pRB + and pRB - cells , cyclin D1 - associated kinase activities were reduced on staurosporine treatment . In contrast , cyclin-dependent kinase ( CDK ) 2 and cyclin E / P24941 REA activities were inhibited only in pRB + cells . DB02010 MEN treatment did not cause reductions in the protein levels of P11802 REA , cyclin D1 , P24941 REA , or cyclin E . The CDK inhibitor proteins P38936 REA ( Waf 1 / Cip 1 ) and p27 ( Kip 1 ) levels increased in staurosporine-treated cells . Immunoprecipitation of P24941 REA , cyclin E , and p2l from staurosporine-treated pRB + cells revealed a 2.5- to 3 - fold higher ratio of p2l bound to P24941 REA compared with staurosporine-treated pRB - cells . In pRB + cells , p2l was preferentially associated with Thrl 6O phosphorylated active P24941 REA . In pRB - cells , however , p2l was bound preferentially to the unphosphorylated , inactive form of P24941 REA even though the phosphorylated form was abundant . This is the first evidence suggesting that P55008 arrest by 4 nM staurosporine is dependent on a functional pRB protein . Cell cycle arrest at the pRB - dependent checkpoint may prevent activation of cyclin E / P24941 REA by stabilizing its interaction with inhibitor proteins p2l and p27 .

Dysregulation of leukocyte gene expression in women with medication-refractory depression versus healthy non-depressed controls . BACKGROUND : Depressive Disorders ( DD ) are a great financial and social burden . Females display 70 % higher rate of depression than males and more than 30 % of these patients do not respond to conventional medications . Thus medication-refractory female patients are a large , under-served , group where new biological targets for intervention are greatly needed . METHODS : We used real-time quantitative polymerase chain reaction ( qPCR ) to evaluate mRNA gene expression from peripheral blood leukocytes for 27 genes , including immune , Q9Y251 REA - axis , ion channels , and growth and transcription factors . Our sample included 23 females with medication refractory DD : 13 with major depressive disorder ( MDD ) , 10 with bipolar disorder ( BPD ) . Our comparison group was 19 healthy , non-depressed female controls . We examined differences in mRNA expression in DD vs . controls , in MDD vs . BPD , and in patients with greater vs . lesser depression severity . RESULTS : DD patients showed increased expression for P22301 REA , P05231 REA , P30559 REA , Q99572 REA , P47900 REA , and Q8NER1 . BPD patients showed increased P05067 REA , P16220 REA , P19838 REA , P04150 REA , and P09486 REA and decreased P01375 REA expression . Depression severity was related to increased P22301 REA , P47900 REA , P51575 REA , and Q9HBA0 expression . CONCLUSIONS : These results support prior findings of dysregulation in immune genes , and provide preliminary evidence of dysregulation in purinergic and other ion channels in females with medication-refractory depression , and in transcription and growth factors in those with BPD . If replicated in future research examining protein levels as well as mRNA , these pathways could potentially be used to explore biological mechanisms of depression and to develop new drug targets .

Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 REA ( P04275 REA ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 REA , plasminogen activator inhibitor type 1 ( P05121 REA ) , antithrombin III ( P01008 REA ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 REA ) ( 1 , 10 , 100 ng / ml ) for up to 48 hours to test the amount of P04275 REA secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 REA are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 REA and P01008 REA difference between FAECs and FVECs . P09038 REA ( 10 ng / ml ) significantly increased P04275 REA secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease .

Possible regulation of migration of intrahepatic cholangiocarcinoma cells by interaction of P61073 REA expressed in carcinoma cells with tumor necrosis factor-alpha and stromal-derived factor - 1 released in stroma . Intrahepatic cholangiocarcinoma ( ICC ) is highly fatal because of early invasion , widespread metastasis , and lack of an effective therapy . We examined roles of P61073 REA and its ligand , stromal cell-derived factor ( SDF ) - 1 , in migration of ICC with respect to tumor-stromal interaction by using two ICC cell lines , a fibroblast cell line ( WI - 38 ) , and 28 human ICC tissues . The two ICC cell lines expressed P61073 REA mRNA and protein , and WI - 38 fibroblasts expressed P48061 REA mRNA and protein . Migration of cultured ICC cells in Matrigel was induced by co-culture with WI - 38 fibroblasts and by incubation with P48061 REA . Anti - P48061 REA antibody suppressed migration , demonstrating that P48061 REA released from WI - 38 fibroblasts was responsible for this migration . P01375 REA ( P01375 REA ) - alpha pretreatment of ICC cells up-regulated P61073 REA mRNA and protein expression in a concentration-dependent manner . Administration of P48061 REA and P01375 REA increased synergistically ICC cell migration , which was suppressed by the P61073 REA antagonist DB06809 MEN . In ICC tissue , P01375 REA was mainly expressed in infiltrated macrophages , P61073 REA in ICC cells , and P48061 REA in stromal fibroblasts . In conclusion , the interaction of P48061 REA released from fibroblasts and P61073 REA expressed on ICC cells may be actively involved in ICC migration , and P01375 REA may enhance ICC cell migration by increasing P61073 REA expression . P61073 REA could be a therapeutic target to prevent ICC invasion .

Amelioration of nephropathy with apoA - 1 mimetic peptide in apoE-deficient mice . BACKGROUND : There is mounting evidence that dyslipidaemia may contribute to development and progression of renal disease . For instance , hyperlipidaemia in apolipoprotein E-deficient ( apoE ( - / - ) ) mice is associated with glomerular inflammation , mesangial expansion and foam cell formation . ApoA - 1 mimetic peptides are potent antioxidant and anti-inflammatory compounds which are highly effective in ameliorating atherosclerosis and inflammation in experimental animals . Given the central role of oxidative stress and inflammation in progression of renal disease , we hypothesized that apoA - 1 mimetic peptide , D - 4F , may attenuate renal lesions in apoE ( - / - ) mice . METHODS : Twenty-five-month-old female apoE ( - / - ) mice were treated with D - 4F ( 300 µg / mL in drinking water ) or placebo for 6 weeks . Kidneys were harvested and examined for histological and biochemical characteristics . RESULTS : Compared with the control mice , apoE ( - / - ) mice showed significant proteinuria , tubulo-interstitial inflammation , mesangial expansion , foam cell formation and up-regulation of oxidative [ NAD ( P ) H oxidase subunits ] and inflammatory [ NF-κB , P13500 REA , P05121 REA and P35354 REA ] pathways . D - 4F administration lowered proteinuria , improved renal histology and reversed up-regulation of inflammatory and oxidative pathways with only minimal changes in plasma lipid levels . CONCLUSIONS : The apoE ( - / - ) mice develop proteinuria and glomerular and tubulo-interstitial injury which are associated with up-regulation of oxidative and inflammatory mediators in the kidney and are ameliorated by the administration of apoA - 1 mimetic peptide . These observations point to the role of oxidative stress and inflammation in the pathogenesis of renal disease in hyperlipidaemic animals and perhaps humans .

Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 REA ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism ( PE ) . tPA has not been replaced by third generation plasminogen activators , e . g . DB00015 ( Ret ) and DB00031 SUB ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor - 1 ( e . g . P05121 REA ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 REA than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility .

G - P04141 REA increases secretion of urokinase-type plasminogen activator by human lung cancer cells . We reported previously that granulocyte colony-stimulating factor ( DB00099 ) can promote the invasion of human lung cancer cell lines in vitro . However , the exact mechanism of its stimulatory effect on invasion remains to be elucidated . In the present study we mainly focused our attention on the components of the plasminogen activation system in human lung cancer cell lines , because of the central role that plasminogen activators play in regulating extracellular proteolysis . We showed that G - P04141 REA induced a dose-dependent increase in the urokinase-type plasminogen activator ( uPA ) activity in the conditioned medium of a PC - 9 lung cancer cell line . When the amounts of uPA activity were quantitated by densitometry , we found that even at a concentration of 0.01 microg / ml , G - P04141 REA had a stimulatory effect on the uPA release , while high concentrations caused a 3.6- fold increase at a maximum concentration of 1 microg / ml . A Western blot analysis of the conditioned medium confirmed the findings observed in a zymographic analysis . The observed increase in uPA protein was paralleled by a significant increase in the uPA mRNA levels after treatment with DB00099 . However , our experiments failed to identify any alteration in the plasminogen activator inhibitor ( P05121 REA ) secretion caused by DB00099 . In addition , we also found the expression of Q99062 REA by PC - 9 cells , suggesting the possible pathway activated by DB00099 .

Oxidative stress-responsive transcription factor P18847 REA potentially mediates diabetic angiopathy . Previous results of our cDNA microarray analysis to look for genes whose expression level correlates well with in vitro tubulogenesis by NP31 endothelial cells revealed the transcription factor P18847 REA known to be responsive to stress such as reactive oxygen species ( ROS ) . Anti - P18847 REA small interfering RNA gave an inhibitory influence on tube formation by NP31 cells expressing an activated form of the vascular endothelial growth factor receptor 1 ( P17948 REA ) kinase . When expression of P18847 REA was regulated under the control of tetracycline system in NP31 cells , they acquired the tubulogenic ability upon P18847 REA induction . While P18847 REA failed to induce expressions of P15692 REA and VEGFR , it regulated those of P24941 REA , P11802 REA , p8 , plasminogen activator inhibitor 1 , integrin alpha 1 , subunit and matrix metalloprotease P45452 REA . In H2O2 - stimulated NP31 cells as well as endothelial cells of glomerulus and aorta of Otsuka-Long-Evans-Tokushima-Fatty diabetic model rats , concomitantly enhanced expressions of P18847 REA , P05121 REA , and p8 were observed . Given the proposed hypothesis of the close linkage between diabetic angiopathy and ROS , those data suggest that ROS-associated diabetic complication may involve P18847 REA - mediated pathological angiogenesis .

Coadministrating luteolin minimizes the side effects of the aromatase inhibitor letrozole . P11511 REA inhibitors ( AIs ) have been used as adjuvant therapeutic agents for breast cancer . Their adverse side effect on blood lipid is well documented . Some natural compounds have been shown to be potential AIs . In the present study , we compared the efficacy of the flavonoid luteolin to the clinically approved AI letrozole ( DB01006 MEN ; Novartis Pharmaceuticals , East Hanover , NJ ) in a cell and a mouse model . In the in vitro experimental results for aromatase inhibition , the Ki values of luteolin and letrozole were estimated to be 2.44 µM and 0.41 nM , respectively . Both letrozole and luteolin appeared to be competitive inhibitors . Subsequently , an animal model was used for the comparison . P11511 REA - expressing MCF - 7 cells were transplanted into ovariectomized athymic mice . Luteolin was given by mouth at 5 , 20 , and 50 mg / kg , whereas letrozole was administered by intravenous injection . Similar to letrozole , luteolin administration reduced plasma estrogen concentrations and suppressed the xenograft proliferation . The regulation of cell cycle and apoptotic proteins-such as a decrease in the expression of Bcl-xL , cyclin-A / D1 / E , P24941 REA / 4 , and increase in that of Bax-was about the same in both treatments . The most significant disparity was on blood lipids . In contrast to letrozole , luteolin increased fasting plasma high-density lipoprotein concentrations and produced a desirable blood lipid profile . These results suggested that the flavonoid could be a coadjuvant therapeutic agent without impairing the action of AIs .

DB00439 MEN , an inhibitor of P04035 REA , inhibits urokinase / urokinase-receptor expression and P14780 REA secretion by peripheral blood monocytes - - a possible protective mechanism against atherothrombosis . It is now recognised that acute myocardial infarction results from the rupture of atherosclerotic plaques . Lymphocytes and macrophages , which infiltrate rupture sites , contribute to plaque degradation by expressing urokinase ( u-PA ) bound to cell membrane by urokinase receptor ( u-PAR ) and by secreting metalloproteinase P14780 REA . We have previously demonstrated that the uptake of oxidised LDL ( ox-LDL ) by monocytes induces an increase of u-PA and u-PAR expression . The present study shows that the expression of u-PA and u-PAR induced by ox-LDL on monocyte surface is suppressed by cerivastatin ( a synthetic inhibitor of P04035 REA , Bayer ) from 2 nM . This leads to reduced plasmin generation and monocyte adhesion to vitronectin . Furthermore , higher concentrations of cerivastatin ( 50-100 nM ) reduce the expression of u-PA and u-PAR on unstimulated monocytes . It also inhibits P14780 REA secretion but has no effect on P01033 REA secretion , suggesting that the decrease in P14780 REA has a real protective effect on plaque stabilisation . The inhibitory effect of cerivastatin on u-PA expression and P14780 REA secretion can be explained by the inhibition of NF-kappa B translocation into the nucleus , as shown by immunofluorescence . As farnesyl-pyrophosphate reverses the effect of cerivastatin , it is postulated that these effects could also be due to the inhibition of Ras prenylation . This was confirmed by confocal microscopy , which shows the Ras delocalisation from the monocyte membrane . The cerivastatin-induced effects on monocyte functions could explain , at least in part , the protective effect of this drug against atherothrombotic events .

The conformational plasticity of calmodulin upon calcium complexation gives a model of its interaction with the oedema factor of Bacillus anthracis . We analyzed the conformational plasticity of calmodulin ( P62158 ) when it is bound to the oedema factor ( EF ) of Bacillus anthracis and its response to calcium complexation with molecular dynamics ( MD ) simulations . The EF - P62158 complex was simulated during 15 ns for three different levels of calcium bound to P62158 . They were respectively no calcium ion ( EF - ( Apo - P62158 ) ) , two calcium ions bound to the C-terminal domain of P62158 ( EF - ( 2Ca - P62158 ) ) , and four calcium ions bound to P62158 ( EF - ( 4Ca - P62158 ) ) . Calculations were performed using AMBER package . The analysis of the MD simulations illustrates how P62158 forces EF in an open conformation to form the adenylyl cyclase enzymatic site , especially with the two calcium form of P62158 , best suited to fit the open conformation of EF . By contrast , P62158 encounters bending and unwinding of its flexible interlinker in EF - ( Apo - P62158 ) and EF - ( 4Ca - P62158 ) . DB01373 MEN binding to one domain of P62158 affects the other one , showing a transmission of information along the protein structure . The analysis of the P62158 domains conformation along the simulations brings an atomistic and dynamic explanation for the instability of these complexes . Indeed the EF-hand helices of the N-terminal domain tend to open upon calcium binding ( EF - ( 4Ca - P62158 ) ) , although the domain is locked by EF . By contrast , the C-terminal domain is strongly locked in the open conformation by EF , and the removal of calcium induces a collapse of EF catalytic site ( EF - ( Apo - P62158 ) ) .

Multiplex protein signature for the detection of bladder cancer in voided urine samples . PURPOSE : Accurate urine assays for bladder cancer detection would benefit patients and health care systems . Through extensive genomic and proteomic profiling of urine components we previously identified a panel of 8 biomarkers that can facilitate the detection of bladder cancer in voided urine samples . In this study we confirmed this diagnostic molecular signature in a diverse multicenter cohort . MATERIALS AND METHODS : We performed a case-control , phase II study in which we analyzed voided urine from 102 subjects with bladder cancer and 206 with varying urological disorders . The urinary concentration of 8 biomarkers ( P10145 REA , P14780 REA and 10 , P05121 REA , P15692 REA , P03950 , Q16790 REA and P02649 REA ) was assessed by enzyme-linked immunosorbent assay . Diagnostic performance of the panel of tested biomarkers was evaluated using ROCs and descriptive statistical values , eg sensitivity and specificity . RESULTS : Seven of the 8 urine biomarkers were increased in subjects with bladder cancer relative to those without bladder cancer . The 7 biomarkers were assessed in a new model , which had an AUROC of 0.88 ( 95 % CI 0.84- 0.93 ) , and 74 % sensitivity and 90 % specificity . In contrast , the sensitivity of voided urine cytology and the UroVysion ® cytogenetic test in this cohort was 39 % and 54 % , respectively . Study limitations include analysis performed on banked urine samples and the lack of voided urine cytology and cytogenetic test data on controls . CONCLUSIONS : The study provides further evidence that the reported panel of diagnostic biomarkers can reliably achieve the noninvasive detection of bladder cancer with higher sensitivity than currently available urine based assays .

The effects of pharmacologic plasminogen activator inhibitor - 1 inhibition in acute and chronic rejection in murine cardiac allografts . BACKGROUND : Acute rejection and graft arterial disease ( Q99259 REA ) in cardiac transplantation limit the long-term survival of recipients ; these processes are enhanced by inflammation and thrombus formation . P00747 REA activator inhibitor ( P05121 REA ) - 1 is critical in the inflammation and thrombus formation . However , little is known about the effect of P05121 REA in heart transplantation . Thus , the objective was to clarify the role of P05121 REA in the progression of cardiac rejection . METHODS : Murine hearts were heterotopically transplanted using major mismatch combinations for evaluation of acute rejection and class II mismatch combinations for the Q99259 REA . We administered the specific P05121 REA inhibitor ( IMD - 1622 ) into murine recipients after cardiac allografts . RESULTS : Nontreated allografts of the major mismatch group were acutely rejected , whereas the P05121 REA inhibitor prolonged their survival . Although severe cell infiltration and intimal thickening with enhancement of inflammatory factors were observed in untreated allografts of class II mismatch group on day 60 , the P05121 REA inhibitor attenuated these changes . CONCLUSION : The P05121 REA inhibitor is potent for the suppression of both acute rejection and Q99259 REA .

Candidate genetic markers and the risk of restenosis after coronary angioplasty . The aim of the present study was to test for possible associations between candidate gene polymorphisms and the risk of restenosis and recurrent restenosis after percutaneous transluminal coronary angioplasty ( PTCA ) without stenting . We followed up 511 PTCA patients , and restenosis and recurrent restenosis were defined according to angiographical criteria . Genotyping of the beta-fibrinogen - 455 G / A , glycoprotein ( GP ) IIIa PlA 1 / PlA 2 , plasminogen activator inhibitor - 1 ( P05121 REA ) 4G / 5G , factor V Leiden 1691 G / A , tumour necrosis factor alpha ( TNFalpha ) - 238 G / A , TNFalpha - 308 G / A , interleukin ( IL ) - 1alpha - 889 C / T , IL - 1beta - 511 C / T , methylenetetrahydrofolate reductase ( P42898 REA ) 677 C / T and endothelial nitric oxide synthase ( P29474 REA ) 4 b / a gene polymorphisms was performed by PCR and restriction-fragment-length-polymorphism-based techniques . One hundred and sixty patients ( 31.3 % ) developed restenosis and in 130 of these patients , of whom 123 were available for analysis , a second PTCA without stenting was performed . Of these patients , 35 ( 28.5 % ) developed recurrent restenosis . None of the investigated genotypes were associated with the risk of restenosis or recurrent restenosis after PTCA . The degree of stenosis before and immediately after PTCA and the severity of the lesion were independent predictors for restenosis after PTCA . In conclusion , there was no association between the beta-fibrinogen - 455 G / A , GP IIIa PlA 1 / A2 , P05121 REA 4G / 5G , factor V Leiden 1691 G / A , TNFalpha - 238 G / A , TNFalpha - 308 G / A , IL - 1alpha - 889 C / T , the IL - 1beta - 511 C / T , P42898 REA 677 C / T and P29474 REA 4 b / a gene polymorphisms and the risk of restenosis after PTCA as well as recurrent restenosis after repeated PTCA .

Overexpression of SnoN / SkiL , amplified at the 3q26 . 2 locus , in ovarian cancers : a role in ovarian pathogenesis . High-resolution array comparative genomic hybridization of 235 serous epithelial ovarian cancers demonstrated a regional increase at 3q26 . 2 encompassing SnoN / SkiL , a coregulator of SMAD / TGFbeta signaling . SnoN RNA transcripts were elevated in approximately 80 % of advanced stage serous epithelial ovarian cancers . In both immortalized normal ( TIOSE ) and ovarian carcinoma cell lines ( OVCA ) , SnoN RNA levels were increased by TGFbeta stimulation and altered by LY294002 and JNK II inhibitor treatment suggesting that the PI3K and JNK signaling pathways may regulate TGFbeta-induced increases in SnoN RNA . In TIOSE , SnoN protein levels were reduced 15min post TGFbeta-stimulation , likely by proteosome-mediated degradation . In contrast , in OVCA , SnoN levels were elevated 3h post-stimulation potentially as a result of inhibition of the proteosome . To elucidate the role of SnoN in ovarian tumorigenesis , we explored the effects of both increasing and decreasing SnoN levels . In both TIOSE and OVCA , SnoN siRNA decreased cell growth between 20 and 50 % concurrent with increased P38936 REA levels . In TIOSE , transient expression of SnoN repressed TGFbeta induction of P05121 REA promoters with little effect on the P38936 REA promoter or resultant cell growth . In contrast to the effects of transient expression , stable expression of SnoN in TIOSE led to growth arrest through induction of senescence . Collectively , these results implicate SnoN levels in multiple roles during ovarian carcinogenesis : promoting cellular proliferation in ovarian cancer cells and as a positive mediator of cell cycle arrest and senescence in non-transformed ovarian epithelial cells .