MH_dev_264

Establishment of a double Philadelphia chromosome-positive acute lymphoblastic leukemia-derived cell line , TMD 5 : effects of cytokines and differentiation inducers on growth of the cells . A double Philadelphia chromosome ( Ph ) - positive leukemia cell line with common-B cell phenotype , designated TMD 5 , was established from the blast cells of a patient with double Ph-positive acute lymphoblastic leukemia . TMD 5 cells expressed 190 kDa P11274 REA / P00519 REA chimeric protein and 145 kDa P00519 REA protein . The cells proliferated without added growth factors . Autocrine growth mechanism was not recognized . The addition of growth factors such as DB00099 , GM - P04141 REA , P08700 REA , P05231 REA , or Stem Cell Factor did not affect the growth . Herbimycin A suppressed the growth of TMD 5 cells at the low concentration that did not affect Ph-negative cells . It suppressed tyrosine phosphorylation of intracellular proteins in TMD 5 cells . Dexamethasone and dibutyryl cyclic AMP also suppressed the growth . They , however , did not affect the phosphorylation significantly . Neither all-trans retinoic acid nor interferon-alpha affected the growth . TMD 5 cells , characterized minutely here and rare in that they have double Ph chromosomes , will be a useful tool for the study of Ph-positive leukemia .

P22888 REA mRNA down-regulation is mediated through P29323 REA - dependent induction of RNA binding protein . The ligand-induced down-regulation of LH receptor ( LHR ) expression in the ovaries , at least in part , is regulated by a posttranscriptional process mediated by a specific LH receptor mRNA binding protein ( LRBP ) . The LH-mediated signaling pathways involved in this process were examined in primary cultures of human granulosa cells . Treatment with 10 IU human chorionic gonadotropin ( hCG ) for 12 h resulted in the down-regulation of LHR mRNA expression while producing an increase in LHR mRNA binding to LRBP as well as a 2 - fold increase in LRBP levels . The activation of P27361 REA / 2 pathway in LH-mediated LHR mRNA down-regulation was also established by demonstrating the translocation of P27361 REA / 2 from the cytosol to the nucleus using confocal microcopy . Inhibition of protein kinase A using H - 89 or P27361 REA / 2 by U0126 abolished the LH-induced LHR mRNA down-regulation . These treatments also abrogated both the increases in LRBP levels as well as the LHR mRNA binding activity . The abolishment of the hCG-induced increase in LRBP levels and LHR mRNA binding activity was further confirmed by transfecting granulosa cells with P27361 REA / 2 specific small interfering RNA . This treatment also reversed the hCG-induced down-regulation of LHR mRNA . These data show that LH-regulated P27361 REA / 2 signaling is required for the LRBP-mediated down-regulation of LHR mRNA .

DB00024 / P16473 REA Signaling Suppresses Fatty Acid Synthase ( P49327 REA ) Expression in Adipocytes . DB00024 / P16473 REA signaling plays a role in the regulation of lipid metabolism in adipocytes . However , the precise mechanisms are not known . In the present study , we determined the effect of DB00024 on fatty acid synthase ( P49327 REA ) expression , and explored the underlying mechanisms . In vitro , DB00024 reduced P49327 REA expression in both mRNA and protein levels in mature adipocytes and was accompanied by protein kinase A ( PKA ) activation , DB02527 - response element binding protein ( CREB ) phosphorylation , as well as extracellular signal-regulated kinase 1/2 ( P27361 REA / 2 ) and c-Jun NH2 - terminal kinase ( JNK ) activation . DB00024 - induced downregulation of P49327 REA was partially abolished by inhibition of PKA and P29323 REA , but not JNK . P16473 REA and P49327 REA expression in visceral tissue was significantly increased in C57BL / 6 mice with diet-induced obesity compared with control animals , whereas thyroid P16473 REA expression was normal . These findings suggest that activation of P16473 REA directly inhibits P49327 REA expression in mature adipocytes , possibly mediated by PKA and P29323 REA . In obese animals , this function of P16473 REA seems to be counteracted . The precise mechanisms need further investigation .

A new mechanism of drug resistance in breast cancer cells : fatty acid synthase overexpression-mediated palmitate overproduction . Multidrug resistance is a major problem in successful cancer chemotherapy . Various mechanisms of resistance , such as ABC transporter-mediated drug efflux , have been discovered using established model cancer cell lines . While characterizing a drug-resistant breast cancer cell line , MCF 7 / AdVp 3000 , we found that fatty acid synthase ( P49327 REA ) is overexpressed . In this study , we showed that ectopic overexpression of P49327 REA indeed causes drug resistance and that reducing the P49327 REA expression increased the drug sensitivity in breast cancer cell lines MCF 7 and MDA-MB - 468 but not in the normal mammary epithelial cell line MCF 10A1 . Use of P49327 REA inhibitor , DB01083 MEN , at low concentrations also sensitized cells with P49327 REA overexpression to anticancer drugs . The P49327 REA - mediated drug resistance appears to be due to a decrease in drug-induced apoptosis from an overproduction of palmitic acid by P49327 REA . Together with previous findings of P49327 REA as a poor prognosis marker for breast cancer patients , our results suggest that P49327 REA overexpression is a new mechanism of drug resistance and may be an ideal target for chemosensitization in breast cancer chemotherapy .

Regulation of P29474 REA - derived superoxide by endogenous methylarginines . The endogenous methylarginines , asymmetric dimethylarginine ( DB01686 MEN ) and N ( G ) - monomethyl - l-arginine ( L-NMMA ) regulate nitric oxide ( NO ) production from endothelial NO synthase ( P29474 REA ) . Under conditions of tetrahydrobiopterin ( BH 4 ) depletion P29474 REA also generates ( * ) O 2 ( - ) ; however , the effects of methylarginines on P29474 REA - derived ( * ) O 2 ( - ) generation are poorly understood . Therefore , using electron paramagnetic resonance spin trapping techniques we measured the dose-dependent effects of DB01686 MEN and L-NMMA on ( * ) O 2 ( - ) production from P29474 REA under conditions of BH 4 depletion . In the absence of BH 4 , DB01686 MEN dose-dependently increased NOS-derived ( * ) O 2 ( - ) generation , with a maximal increase of 151 % at 100 microM DB01686 MEN . L-NMMA also dose-dependently increased NOS-derived ( * ) O 2 ( - ) , but to a lesser extent , demonstrating a 102 % increase at 100 microM L-NMMA . Moreover , the native substrate l-arginine also increased P29474 REA - derived ( * ) O 2 ( - ) , exhibiting a similar degree of enhancement as that observed with DB01686 MEN . Measurements of NADPH consumption from P29474 REA demonstrated that binding of either l-arginine or methylarginines increased the rate of NADPH oxidation . Spectrophotometric studies suggest , just as for l-arginine and L-NMMA , the binding of DB01686 MEN shifts the P29474 REA heme to the high-spin state , indicative of a more positive heme redox potential , enabling enhanced electron transfer from the reductase to the oxygenase site . These results demonstrate that the methylarginines can profoundly shift the balance of NO and ( * ) O 2 ( - ) generation from P29474 REA . These observations have important implications with regard to the therapeutic use of l-arginine and the methylarginine-NOS inhibitors in the treatment of disease .

Retinoic acid and synthetic analogs differentially activate retinoic acid receptor dependent transcription . We have developed an assay where the potency of retinoids in retinoic acid receptor ( RAR ) mediated transcriptional activation can be rapidly evaluated . In this assay hRAR-alpha , hRAR-beta and hRAR-gamma were expressed in CV - 1 cells together with a reporter gene containing a retinoic acid responsive element ( TRE 3 - tk-CAT ) . Concentrations required to obtain half-maximum induction ( ED50 ) of CAT-activity were determined for several retinoids , e . g . , all-trans-retinoic acid ( RA ) , 13 - cis-retinoic acid ( 13 - DB00982 ) , arotinoid acid ( DB02877 MEN ) and m-carboxy-arotinoid acid ( m-carboxy - DB02877 MEN , an inactive arotinoid analog ) . The ED50 values for RA decreased in the order of P10276 REA ( 24 nM ) greater than P10826 REA ( 4.0 nM ) greater than P13631 REA ( 1.3 nM ) , while the ED50 values for DB02877 MEN and 13 - DB00982 decreased in the order of P10276 REA ( 6.5 nM , 190 nM ) greater than P13631 REA ( 2.3 nM , 140 nM ) greater than P10826 REA ( 0.6 nM , 43 nM ) , respectively . No significant inductions were obtained when cells were treated with m-carboxy - DB02877 MEN , even at 10 microM concentrations . The fold induction of CAT-activity for all compounds tested decreased in the order of P10276 REA greater than P10826 REA greater than P13631 REA .

Progressive changes in the leukemogenic signaling in P11274 REA / P00519 REA - transformed cells . Our previous study indicated that P11274 REA / P00519 REA SH2 domain and P11274 REA / P00519 REA SH3 domain + SH2 domain complex are required for immediate activation of the phosphatidylinositol - 3 kinase PI - 3k ) --> Akt serine / threonine kinase pathway and of the signal transducer and activator of transcription 5 ( P42229 REA ) , respectively , in hematopoietic cells . We show here that the defect in activation of PI - 3k / Akt by P11274 REA / P00519 REA DeltaSH 2 mutant ( SH2 domain deleted ) and of P42229 REA by P11274 REA / P00519 REA DeltaSH 3 + DeltaSH 2 mutant ( SH3 and SH2 domains deleted ) is not permanent and both Akt and P42229 REA could be ' re-activated ' by in vitro culture . This phenomenon was responsible for increased resistance to apoptosis , growth factor-independent proliferation and leukemogenesis in SCID mice . Incubation of cells with P11274 REA / P00519 REA tyrosine kinase inhibitor STI 571 abrogated the ' re-activation ' of Akt or P42229 REA by P11274 REA / P00519 REA SH3 + SH2 mutants in some clones , in the others Akt and P42229 REA activation became independent on P11274 REA / P00519 REA kinase activity . The immediate upstream activators of Akt and P42229 REA such as PI - 3k and Jak - 2 were also activated . In addition , the common beta subunit of P08700 REA / P05113 REA / GM - P04141 REA receptor was tyrosine phosphorylated in the clones in which ' re-activation ' was dependent on the P11274 REA / P00519 REA kinase activity . These results suggested that ' re-activation ' of Akt and P42229 REA , in the absence of functional P11274 REA / P00519 REA SH3 + SH2 domains , may be achieved by two different mechanisms : ( i ) P11274 REA / P00519 REA kinase-dependent activation of alternative pathway ( s ) and ( ii ) additional genetic changes stimulating Akt and P42229 REA independently of P11274 REA / P00519 REA . Oncogene ( 2000 ) 19 , 4117 - 4124

Synergism between bosutinib ( DB06616 SUB ) and the Chk 1 inhibitor ( PF - 00477736 ) in highly imatinib-resistant P11274 REA / ABL ⁺ leukemia cells . Interactions between the dual P11274 REA / P00519 REA and Src inhibitor bosutinib and the Chk 1 inhibitor PF - 00477736 were examined in P11274 REA / P00519 REA ( + ) leukemia cells , particularly imatinib-resistant cells , including those with the T315I mutation . Bosutinib blocked PF - 00477736 - induced P27361 REA / 2 activation and sharply increased apoptosis in association with Mcl - 1 inhibition , p34 ( cdc 2 ) dephosphorylation , BimEL up-regulation , and DNA damage in imatinib-resistant CML or Ph ( + ) ALL cell lines . Inhibition of Src or Q02750 REA by shRNA significantly enhanced PF - 0047736 lethality . Bosutinib / PF - 00477736 co-treatment also potentiated cell death in P28906 REA ( + ) CML patient samples , including dasatinib-resistant blast crisis cells exhibiting both T315I and E355G mutations , but was minimally toxic to normal P28906 REA ( + ) cells . Finally , combined in vivo treatment significantly suppressed BaF 3 / T315I tumor growth and prolonged survival in an allogeneic mouse model . Together , these findings suggest that this targeted combination strategy warrants attention in IM-resistant CML or Ph ( + ) ALL .

P22303 REA antagonist potentiated insulin action in fed but not fasted state . The glucose disposal effect of insulin is doubled in response to a meal . This meal-induced insulin sensitization results from insulin acting on the liver , in the presence of a permissive hepatic parasympathetic feeding signal and elevated hepatic glutathione ( DB00143 ) , to release hepatic insulin-sensitizing substance ( HISS ) , a hormone that acts selectively on skeletal muscle to stimulate insulin-mediated glucose uptake . Blockade of the parasympathetic feeding signal to the liver , either through surgical denervation or atropine-mediated antagonism of hepatic muscarinic receptors , eliminates the HISS response , resulting in HISS-dependent insulin resistance ( HDIR ) and decreasing the response to insulin by approximately 55 % in the fed state . P01308 REA action in Sprague-Dawley rats , as determined with a rapidly sampled , transient euglycemic clamp in response to insulin ( 50 mU / kg ) , is decreased in a dose-dependent manner by atropine . In this study , we have used the ED75 atropine-induced model of HDIR . After a submaximal dose of atropine , potentiation of the remaining parasympathetic effect with the acetylcholinesterase antagonist neostigmine significantly restored postprandial insulin sensitization in a dose-dependent manner with peak effect at 0.1 microg / kg / min . DB01400 MEN reversed the insulin resistance induced by partial fasting and partial muscarinic inhibition ( hepatic DB00143 levels are at fed levels ) , but not that induced by surgical hepatic denervation ( DB00143 normal , no nerve signal ) or 24 - h fasting ( low DB00143 ) . No potentiation of the response to insulin by neostigmine occurred in normal , fed rats . The data suggest the use of either direct or indirectly acting cholinergic agonists for the treatment of impaired postprandial insulin sensitization .

DB00796 MEN inhibits Toll-like receptor expression and activity both in vitro and in vivo . INTRODUCTION : Toll-like receptors play an important role in the innate immune system and are found to be crucial in severe diseases like sepsis , atherosclerosis , and arthritis . O60603 REA and O00206 REA expression is upregulated in the inflammatory diseases . Angiotensin II in addition to stimulating vasoconstriction also induces an increase in ROS and a proinflammatory phenotype via AT ( 1 ) R . P30556 REA blocker ( ARB ) , widely used as an antihypertensive drug , has been reported to also have anti-inflammatory effects . Thus , we investigated whether an ARB exerts anti-inflammatory effects via inhibiting O60603 REA and O00206 REA expression . METHODS AND RESULTS : Monocytes were isolated from healthy human volunteers and treated with the synthetic lipoprotein Pam 3CSK4 or LPS in the absence or presence of candesartan . Pretreatment of human monocytes with candesartan significantly decreased Pam 3CSK4 or LPS induced O60603 REA and O00206 REA expression of both mRNA and protein levels ( P < 0.05 vs . control ) along with decrease in the activity of NF-kappaB and the expression of IL - 1beta , P05231 REA , P01375 REA , and P13500 REA . Furthermore , candesartan treated mice show decreased O60603 REA and O00206 REA expression compared to vehicle control mice . CONCLUSION : Pam 3CSK4 and LPS induced O60603 REA and O00206 REA expression at mRNA and protein levels are inhibited by candesartan both in vitro and in vivo . Thus , we define a novel pathway by which candesartan could induce anti-inflammatory effects .

Differential effects of P47900 REA and Q9H244 REA nucleotide receptors on P27361 REA / P28482 REA and phosphatidylinositol 3 - kinase signalling and cell proliferation in serum-deprived and nonstarved glioma P13671 REA cells . We have previously shown that , in glioma P13671 REA cells , two nucleotide ADP-sensitive receptors coexist : P47900 REA , coupled to P98160 REA and responsible for Ca2 + release , and Q9H244 REA , negatively coupled to adenylate cyclase . In the present study , we examined the effects of the stimulation of these two receptors on P27361 REA / 2 and P19957 REA - K activation , and cell proliferation in either serum-deprived or nonstarved P13671 REA cells . In response to ADP and its analogues , in serum-starved cells , both Q8TCB0 P27361 REA and Q8NFH3 P28482 REA were activated in a time-dependent manner , as monitored by Western blot analysis using an antiphospho - Q8NFH3 / Q8TCB0 MAPK antibody . The phosphorylation was reduced both by removal of the extracellular Ca2 + and partially or almost completely by MRS 2179 or AR-C 69931MX , specific antagonists of the P47900 REA and Q9H244 REA receptors , respectively . The inhibitory effect of antagonists was additive . These data indicate the involvement of both receptors , P47900 REA and Q9H244 REA , in the P27361 REA / 2 activation , but the Q9H244 REA receptor contribution predominates . P27361 REA / 2 activity was positively correlated with cell proliferation of cultured glioma P13671 REA cells . In nonstarved cells , ADP markedly decreased the P19957 REA - K activity . In contrast , in serum-starved cells , ADP evoked an increase in the P19957 REA - K activity . Blocking of the P47900 REA receptor by MRS 2179 additionally increased this ADP response . These results suggest that the P47900 REA receptor has an inhibitory and the Q9H244 REA receptor a stimulatory effect on P19957 REA - K signalling pathway . RT-PCR analysis revealed different mRNA expression of both receptors in starved and nonstarved cells . In nonstarved cells , the P47900 REA receptor mRNA predominates , whereas in serum-deprived cells the expression of Q9H244 REA mRNA becomes more pronounced . British Journal of Pharmacology ( 2004 ) 141 , 497-507 . doi : 10.1038 / sj.bjp . 0705639

HRAS 1 and P01308 REA genes are relocated but not structurally altered as a result of the t ( 7 ; 11 ) ( p15 ;p 15 ) in a clone from a patient with acute myeloid leukaemia ( M4 ) . A patient whose leukaemic cells carried the rare t ( 7 ; 11 ) ( p15 ;p 15 ) was diagnosed as having acute myelomonocytic leukaemia ( AML-M 4 ) , and supports the association of this specific translocation with forms of acute myeloid leukaemia showing differentiation . Blast phase chronic myeloid leukaemia was excluded by lack of involvement of the P00519 REA and P11274 REA genes . Chromosome in situ hybridization studies showed that both the HRAS 1 and P01308 REA genes were present on the terminal part of chromosome 11p which was translocated to chromosome 7p . Neither HRAS 1 nor P01308 REA were structurally rearranged . Field inversion gel electrophoresis showed that a 400 kb fragment encompassing HRAS 1 was structurally entire in leukaemic DNA . Because the P01308 REA gene , which was also translocated , is probably located proximal to HRAS 1 on chromosome 11p , it is unlikely that HRAS 1 was near the chromosome 11 breakpoint or involved in this leukaemia .

[ Measurement of rifampicin and clarithromycin in serum by high-performance liquid chromatography with electrochemical detection ] . DB01045 ( RFP ) induces hepatic drug-metabolizing enzymes , making drug interactions a very important clinical problem . DB01211 MENMAX DB01211 MEN ( P62158 ) metabolism is reportedly enhanced by induction of hepatic drug-metabolizing enzymes ( P08684 REA ) by RFP , so that the blood lend of P62158 decreases when RFP is administered concurrently . We connected an electrochemical detector to a high-performance liquid chromatograph ( HPLC ) for simple , rapid , easy measurement of blood concentrations of RFP and P62158 . Using samples of patient serum , normal serum , and reference standards , we compared HPLC by an external laboratory and the results of LC / MS / MS analysis with those of this new assay . A strong correlation was seen between our HPLC results and those of the external laboratory in RFP levels ( r = 0.975 , p < 0.01 ) . A strong correlation was also seen between results we obtained for P62158 with the electrochemical detector in this assay and values measured by LC / MS / MS analysis ( r = 0.995 , p < 0.01 ) . Our method enabled simple , rapid measurement of RFP and P62158 by connecting the HPLC and electrochemical detector in tandem . This system was used to modulate dosage during combined therapy with RFP and P62158 . The therapeutic effect for nontuberculous mycobacteriosis is expected to improve , and our HPLC is expected to be useful for simple , rapid , easy measurement of blood concentrations .

Gateways to clinical trials . 11D10 , 9vPnC - MnCc ; DB00051 , DB00718 , DB00092 , ALN-RSV 01 , AME - 133 , Q99217 REA - 317 , DB00992 , Amlodipine besylate / atorvastatin calcium , Anisodamine , Anti - P05113 REA receptor antibody , Apremilast , Aripiprazole , Atacicept , DB01072 sulfate , Atrasentan ; DB04975 , DB00112 , BIBW - 2992 , DB04853 , BMS - 387032 ; cAC 10 , Caldaret hydrate , CD-NP , DB04918 medocaril , Celivarone fumarate , DB08904 , Cholesteryl hydrophobized polysaccharide-Her 2 protein complex , Choline fenofibrate , Cilengitide , Cinaciguat , Curcumin , Custirsen sodium , Cypher , CYT - 6091 ; Dalcetrapib , Deforolimus , Desvenlafaxine succinate , DB05297 , DP6 - 001 ; E - 7010 , E75 , Ecogramostim , P01133 REA - P64K , EnvPro , Enzastaurin hydrochloride , DB01175 oxalate , DB00973 , DB00973 / simvastatin ; DB05076 ; Gefitinib , Golimumab , Green tea catechins , GTI - 2040 , GW - 406381 ; HPV 16 E6 E7 , HPV -16/18 AS04 , HPV -6/11 / 16/18 ; ICC - 1132 , Immune globulin intravenous ( human ) , DB05039 , Intranasal insulin ; Kahalalide F ; Lactobacillus rhamnosus , Laromustine , Laropiprant , GTI - 2040 ; MAb 3H1 , DB06612 MEN , Mifamurtide , Milataxel , DB05313 ; Nebicapone , DB01280 , Neuradiab ; Oncolytic HSV ; PCV 7 , PHX - 1149 , Pimecrolimus , DB06813 , Pramiconazole ; DB01270 , Reolysin , DB06372 , Rolofylline , DB06176 ; S - 32865 , Shigella dysenteriae 1 vaccine ; Taranabant , Taxus , DB05657 ; Ustekinumab ; Vitespen ; Zileuton , Zycure .

Disruption of Src function potentiates Chk 1 - inhibitor-induced apoptosis in human multiple myeloma cells in vitro and in vivo . Ras / MEK / P29323 REA pathway activation represents an important compensatory response of human multiple myeloma ( MM ) cells to checkpoint kinase 1 ( Chk 1 ) inhibitors . To investigate the functional roles of Src in this event and potential therapeutic significance , interactions between Src and Chk 1 inhibitors ( eg , P55089 REA - 01 or Chk 1i ) were examined in vitro and in vivo . The dual Src / Abl inhibitors BMS 354825 and DB06616 SUB blocked Chk 1 - inhibitor-induced extracellular signal-regulated kinase 1/2 ( P27361 REA / 2 ) activation , markedly increasing apoptosis in association with BimEL up-regulation , p34 ( cdc 2 ) activation , and DNA damage in MM cell lines and primary CD138 ( + ) MM samples . Loss-of-function Src mutants ( K297R , K296R / Y528F ) or shRNA knock-down of Src prevented the P27361 REA / 2 activation induced by Chk 1 inhibitors and increased apoptosis . Conversely , constitutively active Ras or mitogen-activated protein kinase / P29323 REA kinase 1 ( Q02750 REA ) significantly diminished the ability of Src inhibitors to potentiate Chk 1 - inhibitor lethality . Moreover , Src / Chk 1 - inhibitor cotreatment attenuated MM-cell production of vascular endothelial growth factor and other angiogenic factors ( eg , P03950 [ angiogenin ] , P01033 REA / 2 [ tissue inhibitor of metalloproteinases 1/2 ] , and RANTES [ regulated on activation normal T-cell expressed and secreted ] ) , and inhibited in vitro angiogenesis . Finally , coadministration of BMS 354825 and P55089 REA - 01 suppressed human MM tumor growth in a murine xenograft model , increased apoptosis , and diminished angiogenesis . These findings suggest that Src kinase is required for Chk 1 - inhibitor-mediated Ras → P27361 REA / 2 signaling activation , and that disruption of this event sharply potentiates the anti-MM activity of Chk 1 inhi-bitors in vitro and in vivo .

Novel real-time polymerase chain reaction assay for simultaneous detection of recurrent fusion genes in acute myeloid leukemia . The recent World Health Organization classification recognizes different subtypes of acute myeloid leukemia ( AML ) according to the presence of several recurrent genetic abnormalities . Detection of these abnormalities and other molecular changes is of increasing interest because it contributes to a refined diagnosis and prognostic assessment in AML and enables monitoring of minimal residual disease . These genetic abnormalities can be detected using single RT-PCR , although the screening is still labor intensive and costly . We have developed a novel real-time RT-PCR assay to simultaneously detect 15 AML-associated rearrangements that is a simple and easily applicable method for use in clinical diagnostic laboratories . This method showed 100 % specificity and sensitivity ( 95 % confidence interval , 91 % to 100 % and 92 % to 100 % , respectively ) . The procedure was validated in a series of 105 patients with AML . The method confirmed all translocations detected using standard cytogenetics and fluorescence in situ hybridization and some additional undetected rearrangements . Two patients demonstrated two molecular rearrangements simultaneously , with P11274 REA - P00519 REA implicated in both , in addition to Q01196 REA - MECOM in one patient and P29590 REA - P10276 REA in another . In conclusion , this novel real-time RT-PCR assay for simultaneous detection of multiple AML-associated fusion genes is a versatile and sensitive method for reliable screening of recurrent rearrangements in AML .

Association of luteinizing hormone receptor ( LHR ) mRNA with its binding protein leads to decapping and degradation of the mRNA in the p bodies . P22888 REA undergoes downregulation during preovulatory DB00044 MEN surge through a post-transcriptional mechanism involving an RNA binding protein designated as LRBP . The present study examined the mechanism by which LRBP induces the degradation of P22888 REA mRNA , specifically the role of decapping of P22888 REA mRNA and the translocation of LRBP-bound P22888 REA mRNA to degradative machinery . Immunoprecipitation of the complex with the 5 ' cap structure antibody followed by real time PCR analysis showed progressive loss of capped P22888 REA mRNA during downregulation suggesting that P22888 REA mRNA undergoes decapping prior to degradation . RNA immunoprecipitation analysis confirmed dissociation of eukaryotic initiation factor 4E from the cap structure , a step required for decapping . Furthermore , RNA immunoprecipitation analysis using antibody against the p body marker protein , Q9NPI6 showed that P22888 REA mRNA was associated with the p bodies , the cytoplasmic foci that contain RNA degradative enzymes and decapping complex . Immunohistochemical studies using antibodies against LRBP and Q9NPI6 followed by confocal analysis showed colocalization of LRBP with Q9NPI6 during downregulation . This was further confirmed by co-immunoprecipitation of LRBP with Q9NPI6 . The association of LRBP and P22888 REA mRNA in the p bodies during downregulation was further confirmed by examining the association of a second p body component , rck / p54 , using immunoprecipitation and RNA immunoprecipitation respectively . These data suggest that the association of LRBP with P22888 REA mRNA results in the translocation of the messenger ribonucleoprotein complex to the p bodies leading to decapping and degradation .

Loss of Jak 2 impairs endothelial function by attenuating P04049 REA / Q02750 REA / Sp - 1 signaling along with altered P29474 REA activities . A number of inhibitors have been used to dissect the functional relevance of Jak 2 in endothelial homeostasis , with disparate results . Given that Jak 2 deficiency leads to embryonic lethality , the exact role of Jak 2 in the regulation of postnatal endothelial function is yet to be fully elucidated . We generated a model in which Jak 2 deficiency can be induced by tamoxifen in adult mice . Loss of Jak 2 significantly impaired endothelium-dependent response capacity for vasodilators . Matrigel plug assays indicated a notable decrease in endothelial angiogenic function in Jak 2 - deficient mice . Studies in a hindlimb ischemic model indicated that Jak 2 activity is likely to be a prerequisite for prompt perfusion recovery , based on the concordance of temporal changes in Jak 2 expression during the course of ischemic injury and perfusion recovery . A remarkable delay in perfusion recovery , along with reduced capillary and arteriole formation , was observed in Jak 2 - deficient mice . Antibody array studies indicated that loss of Jak 2 led to repressed P29474 REA expression . In mechanistic studies , Jak 2 deficiency attenuated P04049 REA / Q02750 REA signaling , which then reduced activity of Sp - 1 , an essential transcription factor responsible for P29474 REA expression . These data are important not only for understanding the exact role that Jak 2 plays in endothelial homeostasis , but also for assessing Jak 2 - based therapeutic strategies in a variety of clinical settings .

The influence of variation in the Q9H244 REA receptor gene on in vitro platelet inhibition with the direct Q9H244 REA antagonist cangrelor . Novel Q9H244 REA inhibitors are in development to overcome the occurrence of atherothrombotic events associated with poor responsiveness to the widely used Q9H244 REA inhibitor clopidogrel . DB06441 MEN is an intravenously administered Q9H244 REA inhibitor that does not need metabolic conversion to an active metabolite for its antiplatelet action , and as a consequence exhibits a more potent and consistent antiplatelet profile as compared to clopidogrel . It was the objective of this study to determine the contribution of variation in the Q9H244 REA receptor gene to platelet aggregation after in vitro partial Q9H244 REA receptor blockade with the direct antagonist cangrelor . Optical aggregometry was performed at baseline and after in vitro addition of 0.05 and 0.25 microM cangrelor to the platelet-rich plasma of 254 healthy subjects . Five haplotype-tagging ( ht ) - SNPs covering the entire Q9H244 REA receptor gene were genotyped ( rs6798347C > t , rs6787801T > c , rs9859552C > a , rs6801273A > g and rs2046934T > c [ T744C ] ) and haplotypes were inferred . The minor c allele of SNP rs6787801 was associated with a 5 % lower 20 microM ADP-induced peak platelet aggregation ( 0.05 microM cangrelor , p < 0.05 ) . Aa homozygotes for SNP rs9859552 showed 20 % and 17 % less inhibition of platelet aggregation with cangrelor when compared to CC homozygotes ( 0.05 and 0.25 microM cangrelor respectively ; p < 0.05 ) . Results of the haplotype analyses were consistent with those of the single SNPs . Polymorphisms of the Q9H244 REA receptor gene contribute significantly to the interindividual variability in platelet inhibition after partial in vitro blockade with the Q9H244 REA antagonist cangrelor .

Gambogic acid inhibits angiogenesis through suppressing vascular endothelial growth factor-induced tyrosine phosphorylation of P35968 REA / Flk - 1 . Previous studies revealed that gambogic acid ( GA ) , the major active ingredient of gamboge , a brownish to orange resin exuded from Garcinia hanburryi tree in Southeast Asia , possessed significant anticancer activity both in vitro and in vivo . In this study , we explored the high antiangiogenic activities of GA for the first time . GA inhibits the P15692 REA - stimulated proliferation , migration and tube formation of human umbilical vein endothelial cells ( HUVECs ) as well as microvessel sprouting from rat aortic rings in vitro . Moreover , GA inhibits vessel growth in matrigel plugs and P62158 in vivo and transplanted tumor in mice . The results also indicated that GA decreases P15692 REA production of cultured tumor cells and inhibits P15692 REA - induced tyrosine phosphorylation of P35968 REA / Flk - 1 . This inhibition of receptor phosphorylation is correlated with a significant decrease in P15692 REA - triggered phosphorylated forms of P29323 REA , AKT and p38 . Taken together , these findings strongly suggest that GA might be a structurally novel angiogenesis inhibitor .