MH_dev_280

DB02426 MEN effects on brown-fat mitochondria imply that the adenine nucleotide translocator isoforms P12235 REA and P05141 REA may be responsible for basal and fatty-acid-induced uncoupling respectively . In brown-fat mitochondria , fatty acids induce thermogenic uncoupling through activation of P25874 REA ( uncoupling protein 1 ) . However , even in brown-fat mitochondria from P25874 REA - / - mice , fatty-acid-induced uncoupling exists . In the present investigation , we used the inhibitor CAtr ( carboxyatractyloside ) to examine the involvement of the ANT ( adenine nucleotide translocator ) in the mediation of this P25874 REA - independent fatty-acid-induced uncoupling in brown-fat mitochondria . We found that the contribution of ANT to fatty-acid-induced uncoupling in P25874 REA - / - brown-fat mitochondria was minimal ( whereas it was responsible for nearly half the fatty-acid-induced uncoupling in liver mitochondria ) . As compared with liver mitochondria , brown-fat mitochondria exhibit a relatively high ( P25874 REA - independent ) basal respiration ( ' proton leak ' ) . Unexpectedly , a large fraction of this high basal respiration was sensitive to CAtr , whereas in liver mitochondria , basal respiration was CAtr-insensitive . Total ANT protein levels were similar in brown-fat mitochondria from wild-type mice and in liver mitochondria , but the level was increased in brown-fat mitochondria from P25874 REA - / - mice . However , in liver , only Ant 2 mRNA was found , whereas in brown adipose tissue , Ant 1 and Ant 2 mRNA levels were equal . The data are therefore compatible with a tentative model in which the P05141 REA isoform mediates fatty-acid-induced uncoupling , whereas the P12235 REA isoform may mediate a significant part of the high basal proton leak in brown-fat mitochondria .

P35372 REA phosphorylation , desensitization , and ligand efficacy . Mu opioid receptors are subject to phosphorylation and desensitization through actions of at least two distinct biochemical pathways : agonist-dependent mu receptor phosphorylation and desensitization induced by a biochemically distinct second pathway dependent on protein kinase C activation ( 1 ) . To better understand the nature of the agonist-induced mu receptor phosphorylation events , we have investigated the effects of a variety of opiate ligands of varying potencies and intrinsic activities on mu receptor phosphorylation and desensitization . Exposure to the potent full agonists sufentanil , dihydroetorphine , etorphine , etonitazine , and [ D-Ala 2 , MePhe 4 , Glyol 5 ] enkephalin ( DAMGO ) led to strong receptor phosphorylation , while methadone , l-alpha-acetylmethadone ( DB01227 MEN ) , morphine , meperidine , DADL , beta-endorphin ( 1-31 ) , enkephalins , and dynorphin A ( 1-17 ) produced intermediate effects . The partial agonist buprenorphine minimally enhanced receptor phosphorylation while antagonists failed to alter phosphorylation . DB00921 and full antagonists each antagonized the enhanced mu receptor phosphorylation induced by morphine or DAMGO . The rank order of opiate ligand efficacies in producing mu receptor-mediated functional desensitization generally paralleled their rank order of efficacies in producing receptor phosphorylation . Interestingly , the desensitization and phosphorylation mediated by methadone and DB01227 MEN were disproportionate to their efficacies in two distinct test systems . This generally good fit between the efficacies of opiates in mu receptor activation , phosphorylation , and desensitization supports the idea that activated receptor / agonist / G-protein complexes and / or receptor conformational changes induced by agonists are required for agonist-induced mu receptor phosphorylation . Data for methadone and DB01227 MEN suggest possible contribution from their enhanced desensitizing abilities to their therapeutic efficacies .

Pharmacological characterization of 3 - [ 3 - tert-butylsulfanyl - 1 - [ 4 - ( 6 - methoxy-pyridin - 3 - yl ) - benzyl ] - 5 - ( pyridin - 2 - ylmethoxy ) - 1H - indol - 2 - yl ] -2,2- dimethyl-propionic acid ( DB05225 MEN ) , a novel selective P09917 REA - activating protein inhibitor that reduces acute and chronic inflammation . Leukotrienes ( LTs ) are proinflammatory lipid mediators synthesized by the conversion of arachidonic acid ( AA ) to P01374 REA ( 4 ) by the enzyme P09917 REA ( P09917 REA ) in the presence of P09917 REA - activating protein ( P20292 REA ) . 3 - [ 3 - tert-Butylsulfanyl - 1 - [ 4 - ( 6 - methoxy-pyridin - 3 - yl ) - benzyl ] - 5 - ( pyridin - 2 - ylmethoxy ) - 1H - indol - 2 - yl ] -2,2- dimethyl-propionic acid ( DB05225 MEN ) is a novel selective P20292 REA inhibitor in development for the treatment of respiratory conditions such as asthma . In a rat ex vivo whole-blood calcium ionophore-induced Q06643 REA ( 4 ) assay , DB05225 MEN ( administered orally at 1 mg / kg ) displayed > 50 % inhibition for up to 6 h with a calculated EC ( 50 ) of approximately 60 nM . When rat lung was challenged in vivo with calcium ionophore , DB05225 MEN inhibited Q06643 REA ( 4 ) and cysteinyl leukotriene ( CysLT ) production with ED ( 50 ) values of 0.8 and 1 mg / kg , respectively . In this model , the EC ( 50 ) derived from plasma DB05225 MEN was approximately 330 nM for inhibition of both Q06643 REA ( 4 ) and CysLT . In an acute inflammation setting , DB05225 MEN displayed dose-dependent inhibition of Q06643 REA ( 4 ) , CysLT , and plasma protein extravasation induced by peritoneal zymosan injection . In a model of chronic lung inflammation using ovalbumin-primed and challenged BALB / c mice , DB05225 MEN reduced the concentrations of eosinophil peroxidase , CysLTs , and interleukin - 5 in the bronchoalveolar lavage fluid . Finally , DB05225 MEN increased survival time in mice exposed to a lethal intravenous injection of platelet-activating factor . In summary , DB05225 MEN is a novel , potent and selective P20292 REA inhibitor that has excellent pharmacodynamic properties in vivo and is effective in animal models of acute and chronic inflammation and in a model of lethal shock .

Hypoxia-independent angiogenesis in adipose tissues during cold acclimation . The molecular mechanisms of angiogenesis in relation to adipose tissue metabolism remain poorly understood . Here , we show that exposure of mice to cold led to activation of angiogenesis in both white and brown adipose tissues . In the inguinal depot , cold exposure resulted in elevated expression levels of brown-fat-associated proteins , including uncoupling protein - 1 ( P25874 REA ) and P20142 - 1alpha . Proangiogenic factors such as P15692 REA were upregulated , and endogenous angiogenesis inhibitors , including thrombospondin , were downregulated . In wild-type mice , the adipose tissues became hypoxic during cold exposure ; in P25874 REA ( - / - ) mice , hypoxia did not occur , but , remarkably , the augmented angiogenesis was unaltered and was thus hypoxia independent . Intriguingly , P35968 REA blockage abolished the cold-induced angiogenesis and significantly impaired nonshivering thermogenesis capacity . Unexpectedly , P17948 REA blockage resulted in the opposite effects : increased adipose vascularity and nonshivering thermogenesis capacity . Our findings have conceptual implications concerning application of angiogenesis modulators for treatment of obesity and metabolic disorders .

DB06589 SUB inhibits the activation of P09619 REA β-expressing astrocytes in the brain metastatic microenvironment of breast cancer cells . Brain metastases occur in more than one-third of metastatic breast cancer patients whose tumors overexpress P04626 REA or are triple negative . Brain colonization of cancer cells occurs in a unique environment , containing microglia , oligodendrocytes , astrocytes , and neurons . Although a neuroinflammatory response has been documented in brain metastasis , its contribution to cancer progression and therapy remains poorly understood . Using an experimental brain metastasis model , we characterized the brain metastatic microenvironment of brain tropic , P04626 REA - transfected MDA-MB - 231 human breast carcinoma cells ( 231 - BR - P04626 REA ) . A previously unidentified subpopulation of metastasis-associated astrocytes expressing phosphorylated platelet-derived growth factor receptor β ( at tyrosine 751 ; p751 - P09619 REA β ) was identified around perivascular brain micrometastases . p751 - P09619 REA β ( + ) astrocytes were also identified in human brain metastases from eight craniotomy specimens and in primary cultures of astrocyte-enriched glial cells . Previously , we reported that pazopanib , a multispecific tyrosine kinase inhibitor , prevented the outgrowth of 231 - BR - P04626 REA large brain metastases by 73 % . Here , we evaluated the effect of pazopanib on the brain neuroinflammatory microenvironment . DB06589 SUB treatment resulted in 70 % ( P = 0.023 ) decrease of the p751 - P09619 REA β ( + ) astrocyte population , at the lowest dose of 30 mg / kg , twice daily . Collectively , the data identify a subpopulation of activated astrocytes in the subclinical perivascular stage of brain metastases and show that they are inhibitable by pazopanib , suggesting its potential to prevent the development of brain micrometastases in breast cancer patients .

Dynamic regulation of platelet-derived growth factor receptor α expression in alveolar fibroblasts during realveolarization . Although the importance of platelet-derived growth factor receptor ( P09619 REA ) - α signaling during normal alveogenesis is known , it is unclear whether this signaling pathway can regulate realveolarization in the adult lung . During alveolar development , P09619 REA - α-expressing cells induce α smooth muscle actin ( α-SMA ) and differentiate to interstitial myofibroblasts . Fibroblast growth factor ( FGF ) signaling regulates myofibroblast differentiation during alveolarization , whereas peroxisome proliferator-activated receptor ( Q07869 REA ) - γ activation antagonizes myofibroblast differentiation in lung fibrosis . Using left lung pneumonectomy , the roles of FGF and Q07869 REA - γ signaling in differentiation of myofibroblasts from P09619 REA - α-positive precursors during compensatory lung growth were assessed . FGF receptor ( FGFR ) signaling was inhibited by conditionally activating a soluble dominant-negative P21802 REA transgene . Q07869 REA - γ signaling was activated by administration of rosiglitazone . Changes in α-SMA and P09619 REA - α protein expression were assessed in P09619 REA - α-green fluorescent protein ( GFP ) reporter mice using immunohistochemistry , flow cytometry , and real-time PCR . Immunohistochemistry and flow cytometry demonstrated that the cell ratio and expression levels of P09619 REA - α-GFP changed dynamically during alveolar regeneration and that α-SMA expression was induced in a subset of P09619 REA - α-GFP cells . Expression of a dominant-negative P21802 REA and administration of rosiglitazone inhibited induction of α-SMA in P09619 REA - α-positive fibroblasts and formation of new septae . Changes in gene expression of epithelial and mesenchymal signaling molecules were assessed after left lobe pneumonectomy , and results demonstrated that inhibition of P21802 REA signaling and increase in Q07869 REA - γ signaling altered the expression of Shh , FGF , Wnt , and Bmp 4 , genes that are also important for epithelial-mesenchymal crosstalk during early lung development . Our data demonstrate for the first time that a comparable epithelial-mesenchymal crosstalk regulates fibroblast phenotypes during alveolar septation .

Identification of the P07947 REA Kinase as a Therapeutic Target in Basal-Like Breast Cancers . Normal cellular behavior can be described as a complex , regulated network of interaction between genes and proteins . Targeted cancer therapies aim to neutralize specific proteins that are necessary for the cancer cell to remain viable in vivo . Ideally , the proteins targeted should be such that their downregulation has a major impact on the survival / fitness of the tumor cells and , at the same time , has a smaller effect on normal cells . It is difficult to use standard analysis methods on gene or protein expression levels to identify these targets because the level thresholds for tumorigenic behavior are different for different genes / proteins . We have developed a novel methodology to identify therapeutic targets by using a new paradigm called " gene centrality . " The main idea is that , in addition to being overexpressed , good therapeutic targets should have a high degree of connectivity in the tumor network because one expects that suppression of its expression would affect many other genes . We propose a mathematical quantity called " centrality , " which measures the degree of connectivity of genes in a network in which each edge is weighted by the expression level of the target gene . Using our method , we found that several P12931 REA proto-oncogenes P07948 REA , P07947 REA , P08631 REA , P06241 REA , and P06239 REA have high centrality in identifiable subsets of basal-like and P04626 REA + breast cancers . To experimentally validate the clinical value of this finding , we evaluated the effect of P07947 REA knockdown in basal-like breast cancer cell lines that overexpress this gene . We found that P07947 REA downregulation has a significant effect on the survival of these cell lines . Our results identify P07947 REA as a target for therapeutics in a subset of basal-like breast cancers .

Comparative transcriptional network modeling of three Q07869 REA - α / γ co-agonists reveals distinct metabolic gene signatures in primary human hepatocytes . AIMS : To compare the molecular and biologic signatures of a balanced dual peroxisome proliferator-activated receptor ( Q07869 REA ) - α / γ agonist , aleglitazar , with tesaglitazar ( a dual Q07869 REA - α / γ agonist ) or a combination of pioglitazone ( Pio ; Q07869 REA - γ agonist ) and fenofibrate ( Feno ; Q07869 REA - α agonist ) in human hepatocytes . METHODS AND RESULTS : Gene expression microarray profiles were obtained from primary human hepatocytes treated with EC ( 50 ) - aligned low , medium and high concentrations of the three treatments . A systems biology approach , Causal Network Modeling , was used to model the data to infer upstream molecular mechanisms that may explain the observed changes in gene expression . DB08915 MEN , tesaglitazar and Pio / Feno each induced unique transcriptional signatures , despite comparable core Q07869 REA signaling . Although all treatments inferred qualitatively similar Q07869 REA - α signaling , aleglitazar was inferred to have greater effects on high - and low-density lipoprotein cholesterol levels than tesaglitazar and Pio / Feno , due to a greater number of gene expression changes in pathways related to high-density and low-density lipoprotein metabolism . Distinct transcriptional and biologic signatures were also inferred for stress responses , which appeared to be less affected by aleglitazar than the comparators . In particular , Pio / Feno was inferred to increase Q16236 activity , a key component of the stress response pathway , while aleglitazar had no significant effect . All treatments were inferred to decrease proliferative signaling . CONCLUSIONS : DB08915 MEN induces transcriptional signatures related to lipid parameters and stress responses that are unique from other dual Q07869 REA - α / γ treatments . This may underlie observed favorable changes in lipid profiles in animal and clinical studies with aleglitazar and suggests a differentiated gene profile compared with other dual Q07869 REA - α / γ agonist treatments .

Quantitative immuno-positron emission tomography imaging of P04626 REA - positive tumor xenografts with an iodine - 124 labeled anti - P04626 REA diabody . Positron emission tomography ( PET ) provides an effective means of both diagnosing / staging several types of cancer and evaluating efficacy of treatment . To date , the only U . S . Food and Drug Administration-approved radiotracer for oncologic PET is ( 18 ) F-fluoro-deoxyglucose , which measures glucose accumulation as a surrogate for malignant activity . Engineered antibody fragments have been developed with the appropriate targeting specificity and systemic elimination properties predicted to allow for effective imaging of cancer based on expression of tumor associated antigens . We evaluated a small engineered antibody fragment specific for the P04626 REA receptor tyrosine kinase ( P13671 REA . 5 diabody ) for its ability to function as a PET radiotracer when labeled with iodine - 124 . Our studies revealed P04626 REA - dependent imaging of mouse tumor xenografts with a time-dependent increase in tumor-to-background signal over the course of the experiments . Radioiodination via an indirect method attenuated uptake of radioiodine in tissues that express the Na / I symporter without affecting the ability to image the tumor xenografts . In addition , we validated a method for using a clinical PET / computed tomography scanner to quantify tumor uptake in small-animal model systems ; quantitation of the tumor targeting by PET correlated with traditional necropsy-based analysis at all time points analyzed . Thus , diabodies may represent an effective molecular structure for development of novel PET radiotracers .

P35372 REA and P00533 REA contribute to skin pigmentation differences between Indigenous Americans and Europeans . Contemporary variation in skin pigmentation is the result of hundreds of thousands years of human evolution in new and changing environments . Previous studies have identified several genes involved in skin pigmentation differences among African , Asian , and European populations . However , none have examined skin pigmentation variation among Indigenous American populations , creating a critical gap in our understanding of skin pigmentation variation . This study investigates signatures of selection at 76 pigmentation candidate genes that may contribute to skin pigmentation differences between Indigenous Americans and Europeans . Analysis was performed on two samples of Indigenous Americans genotyped on genome-wide SNP arrays . Using four tests for natural selection - - locus-specific branch length ( LSBL ) , ratio of heterozygosities ( lnRH ) , Tajima ' s D difference , and extended haplotype homozygosity ( EHH ) - - we identified 14 selection-nominated candidate genes ( SNCGs ) . SNPs in each of the SNCGs were tested for association with skin pigmentation in 515 admixed Indigenous American and European individuals from regions of the Americas with high ground-level ultraviolet radiation . In addition to Q71RS6 and Q9UMX9 , genes previously associated with European / non-European differences in skin pigmentation , P35372 REA and P00533 REA were associated with variation in skin pigmentation in New World populations for the first time .

Synergistic proapoptotic effects of the two tyrosine kinase inhibitors pazopanib and lapatinib on multiple carcinoma cell lines . DB06589 SUB and lapatinib are two tyrosine kinase inhibitors that have been designed to inhibit the P15692 REA tyrosine kinase receptors 1 , 2 and 3 ( pazopanib ) , and the P00533 REA and P04626 REA receptors in a dual manner ( lapatinib ) . DB06589 SUB has also been reported to mediate inhibitory effect on a selected panel of additional tyrosine kinases such as P09619 REA and c-kit . Here , we report that pazopanib and lapatinib act synergistically to induce apoptosis of A549 non-small-cell lung cancer cells . Systematic assessment of the kinome revealed that both pazopanib and lapatinib inhibited dozens of different tyrosine kinases and that their combination could suppress the activity of some tyrosine kinases ( such as c - DB00134 ) that were not or only partially affected by either of the two agents alone . We also found that pazopanib and lapatinib induced selective changes in the transcriptome of A549 cells , some of which were specific for the combination of both agents . Analysis of a panel of unrelated human carcinoma cell lines revealed a signature of 52 genes whose up - or downregulation reflected the combined action of pazopanib and lapatinib . Indeed , pazopanib and lapatinib exerted synergistic cytotoxic effects on several distinct non-small-cell lung cancer cells as well as on unrelated carcinomas . Altogether , these results support the contention that combinations of tyrosine kinase inhibitors should be evaluated for synergistic antitumor effects . Such combinations may lead to a ' collapse ' of pro-survival signal transduction pathways that leads to apoptotic cell death .

P55157 REA inhibitor decreases plasma cholesterol levels in P01130 REA - deficient WHHL rabbits by lowering the VLDL secretion . To examine whether a microsomal triglyceride transfer protein ( P55157 REA ) - inhibitor is effective in patients with homozygous familial hypercholesterolemia , we administered ( 2S ) - 2 - cyclopentyl - 2 - [ 4 - [ ( 2,4- dimethyl - 9H - pyrido [ 2,3- b ] indol - 9 - yl ) methyl ] phenyl ] - N - [ ( 1S ) - 2 - hydroxy - 1 - phenylethyl ] ethanamide ( DB04852 MEN ) , a new P55157 REA inhibitor , to low-density lipoprotein ( LDL ) - receptor-deficient Watanabe heritable hyperlipidemic ( WHHL ) rabbits at doses of 3 , 6 , and 12 mg / kg for 4 weeks . In the 12 mg / kg group , the plasma cholesterol and triglyceride levels were decreased by 70 % and 45 % , respectively , and the very low-density lipoprotein ( VLDL ) secretion rate was decreased by 80 % . The composition of newly secreted VLDL was similar in each group . This suggests that DB04852 MEN diminished the number of VLDL particles secreted from the liver . Although the ratio of vitamin E / LDL was not altered by DB04852 MEN , triglyceride accumulation and a decrease in vitamin E were observed in the liver . In conclusion , an inhibition of VLDL secretion led to a decrease of plasma LDL in WHHL rabbits , and P55157 REA inhibitors should have hypolipidemic effects against homozygous familial hypercholesterolemia .

Wild-type O75581 REA inhibits , whereas atherosclerosis-linked LRP 6R611C increases PDGF-dependent vascular smooth muscle cell proliferation . Vascular smooth muscle cell ( VSMC ) proliferation is an important event in atherosclerosis and other vasculopathies . PDGF signaling is a key mediator of SMC proliferation , but the mechanisms that control its activity remain unclear . We previously identified a mutation in P01130 REA - related protein 6 ( O75581 REA ) , O75581 REA ( R611C ) , that causes early atherosclerosis . Examination of human atherosclerotic coronary arteries showed markedly increased expression of O75581 REA and colocalization with PDGF receptor β ( P09619 REA - β ) . Further investigation showed that wild-type O75581 REA inhibits but O75581 REA ( R611C ) promotes VSMC proliferation in response to PDGF . We found that wild-type O75581 REA forms a complex with P09619 REA - β and enhances its lysosomal degradation , functions that are severely impaired in O75581 REA ( R611C ) . Further , we observed that wild-type and mutant O75581 REA regulate cell-cycle activity by triggering differential effects on PDGF-dependent pathways . These findings implicate O75581 REA as a critical modulator of PDGF-dependent regulation of cell cycle in smooth muscle and indicate that loss of this function contributes to development of early atherosclerosis in humans .

Targeted therapies for adrenocortical carcinoma : IGF and beyond . Standard chemotherapy for adrenocortical cancer currently is under evaluation in the context of the recently completed FIRM-ACT evaluating the combination of mitotane with either streptozocin or etoposide , cisplatin , and doxorubicin . New agents are eagerly sought by the ACC community that hopes to make progress against this deadly disease . Investigators have begun to dissect the molecular and genomic context of ACC with a goal of identifying potential novel therapeutic agents . One gene consistently overexpressed in ACC is insulin growth factor type 2 . Targeting its receptor P08069 REA has shown encouraging results in ACC cell lines and against murine xenografts . As a result , clinical trials to evaluate agents targeting the P08069 REA have been done including mitotane and DB05759 MEN ( a monoclonal antibody ) and the GALACCTIC trial that has just completed accrual to evaluate OSI - 906 , a small molecule P08069 REA antagonist . On the horizon are other agents targeting other tyrosine kinases , including P01133 REA and FGF , and novel strategies such as individualized tumor analysis to select treatment .

Activation of the Q969V6 REA / actin signaling pathway induces hormonal escape in estrogen-responsive breast cancer cell lines . P03372 REA alpha ( ERα ) is generally considered to be a good prognostic marker because almost 70 % of ERα-positive tumors respond to anti-hormone therapies . Unfortunately , during cancer progression , mammary tumors can escape from estrogen control , resulting in resistance to treatment . In this study , we demonstrate that activation of the actin / megakaryoblastic leukemia 1 ( Q969V6 REA ) signaling pathway promotes the hormonal escape of estrogen-sensitive breast cancer cell lines . The actin / Q969V6 REA signaling pathway is silenced in differentiated ERα-positive breast cancer MCF - 7 and T47D cell lines and active in ERα-negative P50135 REA - 3522 DB00451 - 2 and MDA-MB - 231 breast cancer cells , which have undergone epithelial-mesenchymal transition . We showed that Q969V6 REA activation in MCF - 7 cells , either by modulating actin dynamics or using Q969V6 REA mutants , down-regulates ERα expression and abolishes E2 - dependent cell growth . Interestingly , the constitutively active form of Q969V6 REA represses PR and P04626 REA expression in these cells and increases the expression of HB - P01133 REA , TGFβ , and amphiregulin growth factors in an E2 - independent manner . The resulting expression profile ( ER - , PR - , P04626 REA - ) typically corresponds to the triple-negative breast cancer expression profile .

Electrostatic steering at acetylcholine binding sites . The electrostatic environments near the acetylcholine binding sites on the nicotinic acetylcholine receptor ( nAChR ) and acetylcholinesterase were measured by diffusion-enhanced fluorescence energy transfer ( DEFET ) to determine the influence of long-range electrostatic interactions on ligand binding kinetics and net binding energy . Changes in DEFET from variously charged Tb3 + - chelates revealed net potentials of - 20 mV at the nAChR agonist sites and - 14 mV at the entrance to the P22303 REA active site , in physiological ionic strength conditions . The potential at the alphadelta-binding site of the nAChR was determined independently in the presence of DB01199 MEN to be - 14 mV ; the calculated potential at the alphagamma-site was approximately threefold stronger than at the alphadelta-site . By determining the local potential in increasing ionic strength , Debye-Hückel theory predicted that the potentials near the nAChR agonist binding sites are constituted by one to three charges in close proximity to the binding site . Examination of the binding kinetics of the fluorescent acetylcholine analog dansyl - P13671 REA - choline at ionic strengths from 12.5 to 400 mM revealed a twofold decrease in association rate . Debye-Hückel analysis of the kinetics revealed a similar charge distribution as seen by changes in the potentials . To determine whether the experimentally determined potentials are reflected by continuum electrostatics calculations , solutions to the nonlinear Poisson-Boltzmann equation were used to compute the potentials expected from DEFET measurements from high-resolution models of the nAChR and P22303 REA . These calculations are in good agreement with the DEFET measurements for P22303 REA and for the alphagamma-site of the nAChR . We conclude that long-range electrostatic interactions contribute -0.3 and - 1 kcal / mol to the binding energy at the nAChR alphadelta - and alphagamma-sites due to an increase in association rates .

The PEPvIII-KLH ( DB05374 MEN ) vaccine in glioblastoma multiforme patients . Conventional therapies for glioblastoma multiforme ( GBM ) fail to target tumor cells exclusively , resulting in non-specific toxicity . Immune targeting of tumor-specific mutations may allow for more precise eradication of neoplastic cells . P00533 REA variant III ( EGFRvIII ) is a tumor-specific mutation that is widely expressed in GBM and other neoplasms and its expression enhances tumorigenicity . This in-frame deletion mutation splits a codon , resulting in a novel glycine at the fusion junction producing a tumor-specific epitope target for cellular or humoral immunotherapy . We have previously shown that vaccination with a peptide that spans the EGFRvIII fusion junction ( PEPvIII-KLH / DB05374 MEN ) is an efficacious immunotherapy in syngeneic murine models . In this review , we summarize our results in GBM patients targeting this mutation in multiple , multi-institutional Phase II immunotherapy trials . These trials demonstrated that a selected population of GBM patients who received vaccines targeting EGFRvIII had an unexpectedly long survival time . Further therapeutic strategies and potential pitfalls of using this approach are discussed .

Lipoteichoic acid induces matrix metalloproteinase - 9 expression via transactivation of PDGF receptors and NF-kappaB activation in rat brain astrocytes . Bacterial infections have been shown to be involved in several inflammatory diseases such as brain inflammation . A major factor for these findings is due to the secretion of pro-inflammatory mediators by host cells triggered by the components released from the bacteria . Among these components , lipoteichoic acid ( P01374 REA ) , a component of Gram-positive bacterial cell wall , has been found to be elevated in cerebrospinal fluid of patients suffering from meningitis . Moreover , increased plasma levels of matrix metalloproteinases ( MMPs ) , in particular P14780 REA , have been observed in patients with brain inflammatory diseases and may contribute to disease pathology . However , the molecular mechanisms underlying P01374 REA - induced P14780 REA expression in rat brain astrocytes ( RBA - 1 cells ) remain poorly defined . Here , the data with zymographic , Western blotting , RT-PCR , and immunofluorescent staining analyses showed that P01374 REA induced P14780 REA expression and activation via a O60603 REA - activated c-Src-dependent transactivation of P09619 REA pathway . Transactivation of P09619 REA led to activation of PI3K / Akt and Q8NFH3 / Q8TCB0 MAPK and then activated the IKK / NF-kappaB cascade . The activated-NF-kappaB translocated into nucleus which bound to kappaB-binding site of P14780 REA promoter , and thereby turned on transcription of P14780 REA . Eventually , upregulation of P14780 REA by P01374 REA enhanced cell migration of astrocytes . Taken together , these results suggested that in RBA - 1 cells , activation of NF-kappaB by a c-Src-dependent PI3K / Akt - Q8NFH3 / Q8TCB0 MAPK activation mediated through transactivation of P09619 REA is essential for P14780 REA gene upregulation induced by P01374 REA . Understanding the regulation of P14780 REA expression and functional changes by P01374 REA / TLR system on astrocytes may provide potential therapeutic targets of Gram-positive bacterial infection in brain disorders .

Identification of a variant in P35968 REA associated with serum P35968 REA and pharmacodynamics of DB06589 SUB . PURPOSE : P15692 REA receptor ( VEGFR ) kinases are important drug targets in oncology that affect function of systemic endothelial cells . To discover genetic markers that affect VEGFR inhibitor pharmacodynamics , we performed a genome-wide association study of serum soluble vascular P35968 REA concentrations [ sVEGFR 2 ] , a pharmacodynamic biomarker for P35968 REA inhibitors . EXPERIMENTAL DESIGN : We conducted a genome-wide association study ( GWAS ) of [ sVEGFR 2 ] in 736 healthy Old Order Amish volunteers . Gene variants identified from the GWAS were genotyped serially in a cohort of 128 patients with advanced solid tumor with baseline [ sVEGFR 2 ] measurements , and in 121 patients with renal carcinoma with [ sVEGFR 2 ] measured before and during pazopanib therapy . RESULTS : rs34231037 ( C482R ) in P35968 REA , the gene encoding sVEGFR 2 was found to be highly associated with [ sVEGFR 2 ] , explaining 23 % of the variance ( P = 2.7 × 10 ( - 37 ) ) . Association of rs34231037 with [ sVEGFR 2 ] was replicated in 128 patients with cancer with comparable effect size ( P = 0.025 ) . Furthermore , rs34231037 was a significant predictor of changes in [ sVEGFR 2 ] in response to pazopanib ( P = 0.01 ) . CONCLUSION : Our findings suggest that genome-wide analysis of phenotypes in healthy populations can expedite identification of candidate pharmacogenetic markers . Genotyping for germline variants in P35968 REA may have clinical utility in identifying patients with cancer with unusual sensitivity to effects of P35968 REA kinase inhibitors .

Hypermorphic mutation of phospholipase C , γ2 acquired in ibrutinib-resistant CLL confers Q06187 REA independency upon B-cell receptor activation . DB09053 MENMAX DB09053 MEN has significantly improved the outcome of patients with relapsed chronic lymphocytic leukemia ( CLL ) . Recent reports attribute ibrutinib resistance to acquired mutations in Bruton agammaglobulinemia tyrosine kinase ( Q06187 REA ) , the target of ibrutinib , as well as the immediate downstream effector phospholipase C , γ2 ( P16885 REA ) . Although the C481S mutation found in Q06187 REA has been shown to disable ibrutinib ' s capacity to irreversibly bind this primary target , the detailed mechanisms of mutations in P16885 REA have yet to be established . Herein , we characterize the enhanced signaling competence , Q06187 REA independence , and surface immunoglobulin dependence of the P16885 REA mutation at R665W , which has been documented in ibrutinib-resistant CLL . Our data demonstrate that this missense alteration elicits Q06187 REA - independent activation after B-cell receptor engagement , implying the formation of a novel Q06187 REA - bypass pathway . Consistent with previous results , P16885 REA ( R665W ) confers hypermorphic induction of downstream signaling events . Our studies reveal that proximal kinases P43405 REA and P07948 REA are critical for the activation of mutant P16885 REA and that therapeutics targeting P43405 REA and P07948 REA can combat molecular resistance in cell line models and primary CLL cells from ibrutinib-resistant patients . Altogether , our results engender a molecular understanding of the identified aberration at P16885 REA and explore its functional dependency on Q06187 REA , P43405 REA , and P07948 REA , suggesting alternative strategies to combat acquired ibrutinib resistance .