interacts_with DB00215?
The opposite effects of P40933 and Q9HBE4 on CLL B cells correlate with differential activation of the JAK/ P35610 and P27361 /2 pathways . The clonal expansion of chronic lymphocytic leukemia ( CLL ) cells requires the interaction with the microenvironment and is under the control of several cytokines . Here , we investigated the effect of P40933 and Q9HBE4 , which are closely related to P60568 and share the usage of the common gamma chain and of its P52333 -associated pathway . We found remarkable differences in the signal transduction pathways activated by these cytokines , which determined different responses in CLL cells . P40933 caused cell proliferation and prevented apoptosis induced by surface IgM cross-linking . These effects were more evident in cells stimulated via surface P25942 , which exhibited increased cell expression of IL-15Ralpha chain and , in some of the cases , also of IL-2Rbeta . Q9HBE4 failed to induce CLL cell proliferation and instead promoted apoptosis . Following cell exposure to P40933 , phosphorylation of P42229 was predominantly observed , whereas , following stimulation with Q9HBE4 , there was predominant P42224 and P40763 activation . Moreover , P40933 but not Q9HBE4 caused an increased phosphorylation of Shc and P27361 /2 . Pharmacological inhibition of P52333 or of MEK , which phosphorylates P27361 /2 , efficiently blocked P40933 -induced CLL cell proliferation and the antiapoptotic effect of this cytokine . The knowledge of the signaling pathways regulating CLL cell survival and proliferation may provide new molecular targets for therapeutic intervention . P15692 directly suppresses activation of T cells from ascites secondary to ovarian cancer via P15692 receptor type 2 . BACKGROUND : Vascular endothelial growth factor action in tumour angiogenesis is well characterised ; nevertheless , it functions as a key element in the promotion of the immune system 's evasion by tumours . We sought to investigate the possible direct effect of P15692 on T-cell activation and through which type of P15692 receptor it exerts this effect on cells isolated from ovarian cancer patients ' ascites . METHODS : T cells isolated from the ascites of ovarian cancer patients were cultured with anti-CD3 and P60568 , with or without P15692 for 14 days and the number of viable T cells was counted . Cytotoxic activity of cultured T cells and expression of P15692 receptor-2 ( P35968 ) , was assayed . RESULTS : The addition of P15692 in cultures significantly reduced the number and proliferation rate of T cells in a dose-dependent manner and CD3(+) T cells expressed P35968 on their surface upon activation . Experiments with specific anti- P35968 antibodies revealed that the direct suppressive effect of P15692 on T-cell proliferation is mediated by P35968 . We also showed that P15692 significantly reduced the cytotoxic activity of T cells . CONCLUSION : Our study showed that ascites-derived T cells secrete P15692 and express P35968 upon activation . Vascular endothelial growth factor directly suppresses T-cell activation via P35968 . Bypassing tumor-specific and bispecific antibodies : triggering of antitumor immunity by expression of anti-FcgammaR scFv on cancer cell surface . We have developed a novel immunostimulatory molecule against tumor cells , composed of an anti-FcgammaRIII ( CD16 ) scFv fused to the platelet-derived growth factor receptor ( P09619 ) transmembrane region . This fusion molecule was stably expressed on the tumor cell surface and retained the ability of the parental antibody to bind soluble CD16 . Tumor cells expressing anti-CD16 scFv triggered the release of P60568 by Jurkat-CD 16/gamma cells and of TNFalpha by monocytes when co-cultured with these cells . Furthermore , NK cells could kill scFv-transfected HLA+ class I H1299 lung carcinoma tumor cells , but not the parental cells , indicating that anti-CD16 scFv tumor expression prevents the killer inhibitory receptor ( P55040 ) -mediated inhibition of NK cell cytotoxicity . This anti-CD16 scFv tumor expression also enhanced tumor phagocytosis by IFNgamma-activated macrophages , a mechanism known to induce a protective long-term adaptative immunity to tumors . In vivo Winn tests performed in SCID mice showed that the expression of anti-CD16 scFv on tumor cells , but not of the negative control anti-phOx scFv , prevented tumor cell growth . Thus , expression of FcR antibodies or other FcR-specific ligands on tumor cells represents a novel and potent antibody-based gene therapy approach , which may have clinical applications in cancer DB09559 , a fully human IgG1 mAb directed against the P00533 for the potential treatment of cancer . DB09559 ( DB05774 ) , under development by ImClone Systems in collaboration with Bristol-Myers Squibb , is a fully human IgG1 mAb targeting the epidermal growth factor receptor ( P00533 ) , for the potential intravenous treatment of cancer , in particular NSCLC . In vitro studies demonstrate that necitumumab inhibits downstream targets in the P00533 pathway ( eg , MAPK ) , which are important for cellular proliferation , differentiation , invasion and metastasis . Furthermore , because necitumumab is an IgG1 construct , it has the potential to induce antibody-dependent cell-mediated cytotoxicity against tumor cells . Preclinical studies indicated that the antitumor activity of necitumumab is either comparable with or superior to that of ImClone 's chimeric anti- P00533 mAb cetuximab . In a phase I clinical trial in patients with advanced solid malignancies , necitumumab displayed nonlinear pharmacokinetic behavior . The toxicity profile of necitumumab is acceptable , with skin toxicity being the most frequently reported adverse event in the phase I and II clinical trials conducted to date . Preliminary data from a phase II clinical trial of necitumumab in combination with chemotherapy for the first-line treatment of advanced colon cancer are promising . Success in the ongoing phase III clinical trials in patients with advanced NSCLC would lead to necitumumab becoming a valuable addition to future therapeutic strategies in oncology . Regulation of hepatic lecithin:retinol acyltransferase activity by retinoic acid receptor-selective retinoids . The microsomal enzyme O95237 esterifies retinol and has been implicated in the hepatic storage of vitamin A . Previously , we showed that hepatic O95237 activity is negligible during vitamin A deficiency and that all-trans-retinoic acid ( all-trans-RA ) rapidly induces the activity of liver O95237 in retinoid-deficient rats . In the present studies , we have examined the ability of natural and synthetic retinoids to induce liver O95237 activity in retinoid-deficient rats . The natural retinoids retinol , all-trans-RA ( 100 microg ) , 9- DB00982 , or equal molar amounts of other retinoids were injected ip and O95237 specific activity was measured in liver homogenates 17-18 h later . In retinoid-deficient rats , liver O95237 activity was extremely low [ 0.13 +/- 0.03 pmol retinyl ester ( RE ) /min/mg liver protein , mean +/- SE ] . The natural retinoids retinol and all-trans-RA strongly induced O95237 activity ( 12.71 +/- 1.09 and 13.10 +/- 1.55 pmol RE/min/mg , respectively ) , whereas 9- DB00982 induced a lower level of O95237 activity ( 3.96 +/- 1.88 pmol RE/min/mg , P < 0.001 vs all-trans-RA ) . The retinoic acid receptor ( RAR ) -selective analog ( RAR pan-agonist ) all-trans-UAB8 and the P10276 -selective retinoid Am580 also strongly induced O95237 activity . In contrast , neither RXR-selective agonists nor retinoids having a retro structure were active . For retinoids with significant P10276 binding activity there was a strong direct correlation between receptor binding in vitro and the ability to induce hepatic O95237 activity in vivo ( r2 = 0.920 ) . These data implicate the RARs in the induction of hepatic O95237 and suggest a predominant role for P10276 -active ligands . Modulation of the P22301 /IL-12 cytokine circuit by interferon-beta inhibits the development of epitope spreading and disease progression in murine autoimmune encephalomyelitis . IFN-beta has been shown to be effective in the treatment of multiple sclerosis ( MS ) . However , the primary mechanism by which IFN-beta mediates its therapeutic effect remains unclear . Recent studies indicate that under defined conditions , IFN-beta may downregulate DC expression of IL-12 . We and others have shown that IFN-beta may also downregulate P22301 . In light of the recently proposed paradigm that an P22301 /IL-12 immunoregulatory circuit controls susceptibility to autoimmune disease , we examined the effect of IFN-beta on the development and behavior of the autoreactive T cell repertoire during experimental autoimmune encephalomyelitis ( EAE ) , an animal model sharing many features with MS . SWXJ mice were immunized with the immunodominant p139-151 determinant of myelin proteolipid protein ( PLP ) , and at onset of EAE were treated every other day with IFN-beta . After eight weeks of treatment , we assessed autoreactivity and observed no significant IFN-beta effect on splenocyte proliferation or splenocyte production of P01579 , P60568 , P05112 , or P05113 in response to the priming determinant used to initiate disease . However , in IFN-beta treated mice , the cytokine profile in response to the priming immunogen was significantly skewed toward an increased production of P22301 and a concurrent decreased production of IL-12 . Moreover , the in vivo modulation of the P22301 /IL-12 immunoregulatory circuit in response to the priming immunogen was accompanied by an aborted development of epitope spreading . Our results indicate that IFN-beta induces a reciprocal modulation of the P22301 /IL-12 cytokine circuit in vivo . This skewed autoreactivity establishes an inflammatory microenvironment that effectively prevents endogenous self-priming thereby inhibiting the progression of disease associated with epitope spreading . Expression and biological activity of O95477 in alveolar epithelial cells . The mechanisms used by alveolar type I pneumocytes for maintenance of the lipid homeostasis necessary to sustain these large squamous cells are unknown . The processes may involve the DB00171 -binding cassette transporter A1 ( O95477 ) , a transport protein shown to be crucial in apolipoprotein A-I ( apoA-I ) -mediated mobilization of cellular cholesterol and phospholipid . Immunohistochemical data demonstrated the presence of O95477 in lung type I and type II cells and in cultured pneumocytes . Type II cells isolated from rat lungs and cultured for 5 days in 10 % serum trans-differentiated toward cells with a type I-like phenotype which reacted with the type I cell-specific monoclonal antibody VIIIB2 . Upon incubation of the type I-like pneumocytes with agents that up-regulate the O95477 gene ( 9-cis-retinoic acid [ 9cRA ] and 22-hydroxycholesterol [ 22-OH , 9cRA/22-OH ] ) , O95477 protein levels were enhanced to maximum levels after 8 to 16 hours and remained elevated for 24 hours . In the presence of apoA-I and 9cRA/22-OH , efflux of radioactive phospholipid and cholesterol from pneumocytes was stimulated 3- to 20-fold , respectively , over controls . Lipid efflux was inhibited by DB01599 . DB02772 density gradient analysis of the media from stimulated cells incubated with apoA-I identified heterogeneous lipid particles that isolated at a density between 1.063 and 1.210 g/ml , with low or high apoA-I content . Thus , pneumocytes with markers for the type I phenotype contained functional O95477 protein , released lipid to apoA-I protein , and were capable of producing particles resembling nascent high-density lipoprotein , indicating an important role for O95477 in the maintenance of lung lipid homeostasis . delta- DB00855 dehydratase ( P13716 ) porphyria : the first case in North America with two novel P13716 mutations . The molecular basis of the enzymatic defect responsible for delta-aminolevulinate dehydratase ( P13716 ) porphyria ( ADP ) was investigated in a 14-year-old male who presented clinical and laboratory findings typical of ADP . Nucleotide sequence analysis of P13716 cDNAs from the proband revealed two novel mutations , a 265G to A base transition ( C1 ) and a 394C to T base transition ( P06681 ) , resulting in amino acid substitutions , Glu89Lys and Cys132Arg , respectively . Both mutations were present within exon 5 of the P13716 gene , and appeared to influence the binding of zinc to the enzyme which is essential for enzyme activity . It was found that the C1 mutation was inherited from his father , while the P06681 mutation was from his mother . Expression of these mutant P13716 cDNAs in Chinese hamster ovary cells produced normal P13716 mRNA levels , but markedly decreased P13716 protein and enzyme activity . These results suggest that the combination of the two aberrant ALADs with little enzyme activity accounts for the markedly decreased P13716 activity observed in the proband . This case represents the molecular analysis of the P13716 gene defects in the first case of ADP identified in North America , who is a compound heterozygote for two novel P13716 gene defects . DB00067 receptor expression and mutation analysis in corticotropin-secreting tumors . DB00067 is an important regulator of hypothalamo-pituitary-adrenal axis activation , primarily acting through the V3 receptor ( V3R ) . Many patients with DB01285 -secreting pituitary adenomas , but not normal individuals , respond to desmopressin , a relatively V2-specific vasopressin agonist , with increased DB01285 and cortisol levels . We have searched for mutations of the V3R gene in DB01285 -secreting pituitary adenomas and one ectopic DB01285 -secreting tumor . No abnormalities were found in 12 tumors studied by PCR-single strand conformation polymorphism ( PCR-SSCP ) analysis . We then verified by RT-PCR whether the response to desmopressin was due to overexpression of the V3R or abnormal expression of the P30518 in the pituitary tumor . We found that the P30518 gene was expressed in a number of corticotroph tumors and in the DB01285 -secreting ectopic tumor , and that the V3R gene appears to be overexpressed in these tumors . We conclude that V3R mutations are unlikely to be present in the DB01285 -secreting tumors we examined , but that the P30518 gene is expressed in the majority of the samples tested , and the V3R is expressed in all of these tumors . We speculate that the response to the desmopressin test observed in patients with Cushing 's disease may be due to abnormal expression of V3R or P30518 in DB01285 -secreting tumors . DB01599 alleviates atherosclerosis and improves high density lipoprotein function . BACKGROUND : DB01599 is a unique hypolipidemic agent that decreases high density lipoprotein cholesterol ( HDL-C ) . However , it is not definite that whether probucol hinders the progression of atherosclerosis by improving HDL function . METHODS : Eighteen New Zealand White rabbits were randomly divided into the control , atherosclerosis and probucol groups . Control group were fed a regular diet ; the atherosclerosis group received a high fat diet , and the probucol group received the high fat diet plus probucol . Hepatocytes and peritoneal macrophages were isolated for [ (3)H ] labeled cholesterol efflux rates and expression of O95477 and SR-B1 at gene and protein levels ; venous blood was collected for serum paraoxonase 1 , myeloperoxidase activity and lipid analysis . Aorta were prepared for morphologic and immunohistochemical analysis after 12 weeks . RESULTS : Compared to the atherosclerosis group , the paraoxonase 1 activity , cholesterol efflux rates , expression of O95477 and Q8WTV0 in hepatocytes and peritoneal macrophages , and the level of O95477 and Q8WTV0 in aortic lesions were remarkably improved in the probucol group , But the serum HDL cholesterol concentration , myeloperoxidase activity , the IMT and the percentage plaque area of aorta were significantly decreased . CONCLUSION : DB01599 alleviated atherosclerosis by improving HDL function . The mechanisms include accelerating the process of reverse cholesterol transport , improving the anti-inflammatory and anti-oxidant functions . Q9UEF7 endows hepatoma cells with resistance to anoikis via P35968 /PAK1 activation in hepatocellular carcinoma . Q9UEF7 was originally characterized as an aging suppressor gene that predisposed Q9UEF7 -deficient mice to premature aging-like syndrome . Although Q9UEF7 was recently reported to exhibit tumor suppressive properties during various malignant transformations , the functional role and molecular mechanism of Q9UEF7 in hepatocarcinogenesis remains poorly understood . In our present study , immunohistochemical Q9UEF7 staining levels in a clinical follow-up of 52 hepatoma patients were significantly associated with liver cirrhosis , tumor multiplicity and venous invasion . The overall survival rate of hepatoma patients with high Q9UEF7 expression was significantly lower than those patients with low Q9UEF7 expression . Moreover , Q9UEF7 overexpression increased cellular migration , anchorage-independent growth , and anoikis resistance in hepatoma cells . Q9UEF7 overexpression elevated Q13153 ( PAK1 ) expression and shRNA-mediated PAK1 knockdown and kinase activity inhibition with kinase dead mutant PAK1 K299R coexpression or allosteric inhibitor IPA3 treatment reversed anoikis resistance in Q9UEF7 -overexpressed hepatoma cells . More importantly , the pivotal significance of upregulated P35968 protein levels mediated by Q9UEF7 expression was confirmed by P35968 inhibitor DB06626 and blocking antibody treatment in hepatoma cells . DB06626 treatment sensitized anoikis was reversed by constitutive active mutant PAK1 T423E coexpression in Q9UEF7 -overexpressed hepatoma cells . Conversely , knockdown of Q9UEF7 reduced P35968 /PAK1 dependent anoikis resistance , which could be reversed by PAK1 T423E . These results revealed a novel oncogenic function of Q9UEF7 in promoting anoikis resistance via activating P35968 /PAK1 signaling , thus facilitating tumor migration and invasion during hepatoma progression , which could provide a putative molecular mechanism for tumor metastasis . [ 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 ( P62158 ) metabolism is reportedly enhanced by induction of hepatic drug-metabolizing enzymes ( P08684 ) 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 . Short-term biomarker modulation prevention study of anastrozole in women at increased risk for second primary breast cancer . The selective estrogen receptor modulators ( SERM ) , Tamoxifen and raloxifen reduce risk breast cancer . Patient acceptance of SERMs for breast cancer prevention is low due to toxicities . New agents with a better toxicity profile are needed . P11511 inhibitors ( AI ) reduce the risk of contralateral breast cancer and risk of new breast cancer in high risk women . However , the mechanism by which AIs reduce breast risk is not known . Surrogate biomarkers are needed to evaluate the effect of preventive agents . The objective of this prospective short-term prevention study was to evaluate the effect of anastrozole on biomarkers in breast tissue and serum of women at increased risk for developing a contralateral breast cancer . Women with a history of stage I , II breast cancer who started anastrozole for standard adjuvant treatment were eligible . Patients underwent baseline fine needle aspiration of the unaffected breast and serum collection for biomarker analysis before starting anastrozole at 1 mg per oral/day and again at 6 months . Biomarkers included changes in cytology , insulin-like growth factor 1 ( DB01277 ) , P08833 ( P08833 ) , and P17936 . Thirty-seven patients were enrolled . There was a significant modulation in serum P08833 levels between pre- and postsamples ( P = 0.02 ) . No change was observed in DB01277 , P17936 , and breast cytology.We showed a significant modulation of P08833 levels with six months anastrozole . DB01217 is currently being studied as a prevention agent in a large phase III trial and our results provide support for continued evaluation of P08833 as a surrogate endpoint biomarker in prospective breast chemoprevention studies . Serotonergic mechanisms in human allergic contact dermatitis . Expression of serotonin ( 5-hydroxytryptamine ; 5-HT ) , 5-HT receptors 1A ( 5-HT1AR ) and 2A , and serotonin transporter protein ( P31645 ) was studied in positive epicutaneous reactions to nickel sulphate in nickel-allergic patients , at 72 h post-challenge with the antigen . In addition , the effects of 5-HT2AR agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , and the selective serotonin reuptake inhibitors ( SSRIs ) citalopram and fluoxetine , were tested on nickel-stimulated peripheral blood mononuclear cells from nickel-allergic patients , regarding their proliferation and interleukin ( IL ) -2 production , as well as the effect of these SSRIs on a murine Langerhans ' cell-like line ( XS52 ) , regarding its IL-1beta production . Serotonin-positive platelets were increased in the inflamed skin compared with control skin . A decrease ( p < 0.01 ) in 5-HT1AR-positive mononuclear cells was evident in the eczematous skin compared with control skin , whereas 5-HT2AR- and P31645 -positive cells were increased ( p < 0.001 for both ) in the eczematous skin . Treatment of nickel-stimulated peripheral blood mononuclear cells with 5x10(-5) mol/l of DOI inhibited ( p < 0.01 ) the proliferation of nickel-stimulated peripheral blood mononuclear cells , while no effect was found regarding P60568 production . DB00215 at 10(-6) mol/l tended to inhibit the production of IL-1beta by the XS52 cell line . These results indicate the implication of the serotonergic system in the contact allergic reaction . DB00067 -induced cytoplasmic and nuclear calcium signaling in embryonic cortical astrocytes : dynamics of calcium and calcium-dependent kinase translocation . The present study sought to determine the downstream consequences of V1a vasopressin receptor ( P37288 ) activation of Ca2+ signaling in cortical astrocytes . Results of these analyses demonstrated that P37288 activation led to a marked increase in both cytoplasmic and nuclear Ca2+ . We also investigated P37288 activation of Ca2+-activated signaling kinases , protein kinase C ( PKC ) , Ca2+/calmodulin-dependent protein kinase II ( CaMKII ) , and the mitogen-activated protein ( Q96HU1 ) kinases [ MAPK and extracellular signal-regulated kinases 1 and 2 ( P27361 /2 ) ] , their localization within cytoplasmic and nuclear compartments , and activation of their downstream nuclear target , the transcription factor DB02527 response element-binding protein ( CREB ) . Results of these analyses demonstrated that P37288 activation led to a significant rise in PKC , CaMKII , and P27361 /2 activation , with CaMKII and P27361 /2 demonstrating dynamic transport between cytoplasmic and nuclear compartments . Although no evidence of PKC translocation was apparent , PKC and CaMKs were required for activation and nuclear translocation of P27361 /2 . Subsequent to CaMKII and P27361 /2 translocation to the nucleus , CREB activation occurred and was found to be dependent on upstream activation of P27361 /2 and CaMKs . These data provide the first systematic analysis of the P37288 -induced Ca2+ signaling cascade in cortical astrocytes . In addition , results of this study introduce a heretofore unknown effect of vasopressin , dynamic Ca2+ signaling between the cytoplasm and nucleus that leads to comparable dynamics of kinase activation and shuttling between cytoplasmic and nuclear compartments . Implications for development and regeneration induced by P37288 activation of CREB-regulated gene expression in cortical astrocytes are discussed . Glucocorticoids regulate calcineurin-dependent trans-activating pathways for interleukin-2 gene transcription in human T lymphocytes . Glucocorticoids ( GC ) inhibit P60568 gene transcription by interfering with the binding of the nuclear factor activator protein-1 on the P60568 promoter . Calcineurin , a Ca2+/calmodulin-dependent protein phosphatase , is an essential component of the T cell antigen receptor signal transduction pathway leading to P60568 gene transcription . Therefore , we have asked whether this phosphatase may also be regulated by GC . Jurkat T cells were cotransfected with plasmids containing the intact P60568 promoter or its NF-AT and P14859 motifs , and a deletion mutant ( delta P62158 -AI ) of calcineurin known to have Ca(2+)-independent constitutive phosphatase activity . Cotransfection of P60568 promoter with delta P62158 -AI allowed the activation of P60568 promoter in the presence of phorbol ester alone . Under these conditions dexamethasone ( DB00514 ; 10(-6) M ) inhibited P60568 promoter activation by 50-60 % . The inhibitory effect of DB00514 was specific , as demonstrated by experiments using an unrelated promoter ( simian virus 40 ) and estradiol . Furthermore , it was completely reversed in the presence of excess amounts of the glucocorticoid antagonist RU 486 , which suggests that it is mediated through the glucocorticoid receptor . Overexpression of calcineurin via delta P62158 -AI in Jurkat cells decreased their apparent sensitivity to DB00514 ( approximately 5-fold increase in IC50 ) . Similar results were obtained with the NF-AT and P14859 constructs , which are also known to be activated by calcineurin . Thus , in addition to their known inhibitory effects on activator protein-1 , GC also inhibit calcineurin-dependent pathways for T cell activation . DB06626 for renal cell carcinoma . BACKGROUND : The approval of sunitinib , sorafenib and temsirolimus has dramatically altered the management of renal cell carcinoma ( RCC ) . DB00112 plus IFN may also be added to the therapeutic armamentarium . DB06626 ( AG-013736 ) is an oral and selective tyrosine kinase inhibitor . OBJECTIVE : Data supporting the development of axitinib for RCC are reviewed . METHODS : Preclinical and clinical data available for axitinib for RCC are presented . RESULTS : DB06626 inhibits P17948 , P35968 and P35916 with picomolar potencies , and P16234 , P09619 and c-kit with nanomolar potencies . Phase II clinical trials of axitinib in pretreated RCC following sorafenib or cytokine treatment have demonstrated promising activity accompanied by a favorable toxicity profile . Further development of axitinib for RCC is warranted . A fully human recombinant IgG-like bispecific antibody to both the epidermal growth factor receptor and the insulin-like growth factor receptor for enhanced antitumor activity . Both the epidermal growth factor receptor ( P00533 ) and the insulin-like growth factor receptor ( IGFR ) have been implicated in the tumorigenesis of a variety of cancers . Here we propose that simultaneous targeting of both receptors with a bispecific antibody would lead to enhanced antitumor activity . To this end , we produced a recombinant human IgG-like bispecific antibody , a Di-diabody , using the variable regions from two antagonistic antibodies : DB05774 to P00533 and DB05759 to IGFR . The Di-diabody binds to both P00533 and IGFR and effectively blocked both P01133 - and IGF-stimulated receptor activation and tumor cell proliferation . The Di-diabody also inherited the biological properties from both of its parent antibodies ; it triggers rapid and significant IGFR internalization and degradation and mediates effective antibody-dependent cellular cytotoxicity in a variety of tumor cells . Finally , the Di-diabody strongly inhibited the growth of two different human tumor xenografts in vivo . Our results underscore the benefits of simultaneous targeting of two tumor targets with bispecific antibodies . P11511 inhibitors and cyclooxygenase-2 ( P35354 ) 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 overexpression has recently been detected in endometriotic tissue . P11511 ( p450arom ) is responsible for converting C19 androgens into estrogen in several types of human tissue . P11511 activity causes local estrogen biosynthesis , which , in turn , stimulates prostaglandin E2 production by upregulating cyclooxygenase-2 ( P35354 ) . 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 are thought to be promising new therapeutic targets . Thus , specific aromatase inhibitors ( e.g. DB01006 / DB01006 , DB01217 /Arimidex or Exemestan/Aromasin ) or selective P35354 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 . Silencing of ALA dehydratase affects ALA-photodynamic therapy efficacy in K562 erythroleukemic cells . Synthesis of protoporphyrin IX ( PpIX ) by malignant cells is essential for the success of ALA-based photodynamic therapy ( PDT ) . Two key enzymes that were described as affecting PpIX accumulation during ALA treatment are porphobilinogen deaminase ( P08397 ) and ferrochelatase . Here , we show that down regulation of ALA dehydratase ( P13716 ) expression and activity by specific shRNA induced a marked decrease in PpIX synthesis in K562 erythroleukemic cells . Photo-inactivation efficacy following DB00855 was directly correlated with P13716 -silencing and cellular levels of PpIX . MTT metabolism following DB00855 was shown to be 60 % higher in P13716 -silenced cells in comparison to control cells , indicating that mitochondria were protected in the silenced cells . Morphological analysis by scanning electron microscopy ( SEM ) of cells treated by DB00855 showed no morphological changes in P13716 -silenced cells , in contrast to controls exhibiting cell deformations and lysis . Membrane integrity following DB00855 was kept intact and undamaged in P13716 -silenced cells as examined by P08758 -FITC/PI staining and LDH-L leakage . We conclude that P13716 , although it is present in the cell at abundant levels , has a major and limiting role in regulating PpIX synthesis and DB00855 outcome . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . Structural basis for P01133 receptor inhibition by the therapeutic antibody DB05774 . Therapeutic anticancer strategies that target and inactivate the epidermal growth factor receptor ( P00533 ) are under intense study in the clinic . Here we describe the mechanism of P00533 inhibition by an antibody drug DB05774 . DB05774 is a fully human antibody that has similar antitumor potency as the chimeric cetuximab/Erbitux and might represent a safer therapeutic alternative . We report the X-ray crystal structure of the Fab fragment of DB05774 ( Fab11F8 ) in complex with the entire extracellular region and with isolated domain III of P00533 . We compare this to our previous study of the cetuximab/ P00533 interaction . Fab11F8 interacts with a remarkably similar epitope , but through a completely different set of interactions . Both the similarities and differences in binding of these two antibodies have important implications for the development of inhibitors that could exploit this same mechanism of P00533 inhibition . DNA methylation differences after exposure to prenatal famine are common and timing- and sex-specific . Prenatal famine in humans has been associated with various later-life consequences , depending on the gestational timing of the insult and the sex of the exposed individual . Epigenetic mechanisms have been proposed to underlie these associations . Indeed , animal studies and our early human data on the imprinted P01344 locus indicated a link between prenatal nutritional and DNA methylation . However , it remains unclear how common changes in DNA methylation are and whether they are sex- and timing-specific paralleling the later-life consequences of prenatal famine exposure . To this end , we investigated the methylation of 15 loci implicated in growth and metabolic disease in individuals who were prenatally exposed to a war-time famine in 1944-45 . Methylation of INSIGF was lower among individuals who were periconceptionally exposed to the famine ( n = 60 ) compared with their unexposed same-sex siblings ( P = 2 x 10(-5) ) , whereas methylation of P22301 , P41159 , O95477 , GNASAS and Q9UI56 was higher ( all P < 10(-3) ) . A significant interaction with sex was observed for INSIGF , P41159 and GNASAS . Next , methylation of eight representative loci was compared between 62 individuals exposed late in gestation and their unexposed siblings . Methylation was different for GNASAS ( P = 1.1 x 10(-7) ) and , in men , P41159 ( P = 0.017 ) . Our data indicate that persistent changes in DNA methylation may be a common consequence of prenatal famine exposure and that these changes depend on the sex of the exposed individual and the gestational timing of the exposure . Neurohypophysial receptor gene expression by thymic T cell subsets and thymic T cell lymphoma cell lines . Neurohypophysial oxytocin ( OT ) and vasopressin ( VP ) genes are transcribed in thymic epithelium , while immature T lymphocytes express functional neurohypophysial receptors . Neurohypophysial receptors belong to the G protein-linked seven-transmembrane receptor superfamily and are encoded by four distinct genes , OTR , V1R , P30518 and V3R . The objective of this study was to identify the nature of neurohypophysial receptor in thymic T cell subsets purified by immunomagnetic selection , as well as in murine thymic lymphoma cell lines RL12-NP and BW5147 . OTR is transcribed in all thymic T cell subsets and T cell lines , while V3R transcription is restricted to P01730 +CD8+ and CD8+ thymic cells . Neither V1R nor P30518 transcripts are detected in any kind of T cells . The OTR protein was identified by immunocytochemistry on thymocytes freshly isolated from C57BL/6 mice . In murine fetal thymic organ cultures , a specific OTR antagonist does not modify the percentage of T cell subsets , but increases late T cell apoptosis further evidencing the involvement of OT/OTR signaling in the control of T cell proliferation and survival . According to these data , OTR and V3R are differentially expressed during T cell ontogeny . Moreover , the restriction of OTR transcription to T cell lines derived from thymic lymphomas may be important in the context of T cell leukemia pathogenesis and treatment . Local immunotherapy of glioma patients with a combination of 2 bispecific antibody fragments and resting autologous lymphocytes : evidence for in situ t-cell activation and therapeutic efficacy . After adoptive transfer of pre-activated lymphocytes into the operation cavity of glioma patients , tumor regression and improved survival have been reported in some patients . Results were most impressive when bispecific antibodies with tumor x CD3 specificity were also applied . In this study , we attempted to avoid time-consuming pre-activation procedures for adoptively transferred cells by using a combination of bispecific antibodies directed to the P01133 receptor ( P00533 ) on tumor cells and to CD3 and P10747 on T cells . Eleven patients with high-grade malignant glioma received 3 injections of 2 bispecific antibody fragments ( P00533 x CD3 and P00533 x P10747 ) together with freshly isolated autologous lymphocytes via an Ommaya reservoir . Intracavitary fluid aspirated during immunotherapy was examined for markers of T-cell activation . Increased levels of soluble P60568 receptor and P01375 were detected in the intracavitary fluid of all patients tested . Two of the 11 treated patients experienced a beneficial response to therapy as defined by a transient contrast enhancement in subsequent Q9BWK5 scans and prolonged survival . Side effects were transient and consisted of fever , nausea , headache and aggravation of pre-existing neurologic deficits . These adverse effects were most likely due to the antibody construct containing anti-CD3 specificity . Two patients developed cerebral edema and required steroid treatment . [ P11511 inhibitors -- theoretical concept and present experiences in the treatment of endometriosis ] . The medical treatment of endometriosis needs to be optimized . Therapeutic management strategies of endometriosis-associated pain or recurrent disease is primarily aimed at downregulating the ovarian function or at antagonizing the effect of estrogen in ectopic endometrial implants . In this context , basic research is delivering powerful tools for the possible development of new , specific treatment modalities . Recently , aromatase overexpression has been detected in endometriotic tissue . P11511 ( p450arom ) is responsible for conversion of C19 androgens to estrogen in several human tissues . P11511 activity gives rise to local estrogen biosynthesis , which , in turn , stimulates prostaglandin E(2) production by upregulation of cyclooxygenase-2 ( P35354 ) , thus establishing a positive feedback cycle . Another abnormality in endometriosis , i. e. the deficiency in 17 beta-hydroxysteroiddehydrogenase type-II ( 17 beta-HSD-Type-II ) expression , impairs the inactivation of estradiol to estrone . In contrast to the eutopic endometrium , these molecular aberrations collectively favour accumulation of increasing amounts of local estradiol and prostaglandin E(2) 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 are promising new therapeutic targets . In summary , specific aromatase inhibitors ( such as Letrozole , DB01217 or Exemestan ) or selective P35354 inhibitors ( e.g. Celecoxib , DB00533 ) are of great interest to be studied in clinical trials in premenopausal woman with endometriosis to extend the spectrum of currently available treatment options . Retinoic acid-receptor activation of P07988 gene transcription in respiratory epithelial cells . Retinoids are known to play important roles in organ development of the lung . Retinoids exert their activity by modulating the expression of numerous genes , generally influencing gene transcription , in target cells . In the present work , the mechanism by which retinoic acid ( RA ) regulates surfactant protein ( SP ) B expression was assessed in vitro . RA ( 9- DB00982 ) enhanced P07988 mRNA in pulmonary adenocarcinoma cells ( H441 cells ) and increased transcriptional activity of the P07988 promoter in both H441 and mouse lung epithelial cells ( MLE-15 ) . Cotransfection of H441 cells with retinoid nuclear receptor ( RAR ) -alpha , -beta , and -gamma and retinoid X receptor ( RXR ) -gamma further increased the response of the P07988 promoter to RA . Treatment of H441 cells with RA increased immunostaining for the P07988 proprotein and increased the number of cells in which the P07988 proprotein was detected . An RA responsive element mediating RA stimulation of the human P07988 promoter was identified . P10276 and -gamma and RXR-alpha but not P10826 or RXR-beta and -gamma were detected by immunohistochemical analysis of H441 cells . RA , by activating RAR activity , stimulated the transcription and synthesis of P07988 in pulmonary adenocarcinoma cells . DB01599 inhibits O95477 -mediated cellular lipid efflux . OBJECTIVE : DB00171 -binding cassette transporter A1 ( O95477 ) mediates the efflux of lipids from cells to lipid-poor apolipoproteins . In this article , we characterize the effect of probucol on cellular O95477 -mediated lipid efflux . METHODS AND RESULTS : DB01599 inhibited cholesterol efflux up to 80 % in J774 macrophages expressing O95477 . In Fu5AH hepatoma cells that contain scavenger receptor class B , type I , but not functional O95477 , we observed no effect of probucol on cholesterol efflux . DB01599 inhibited cholesterol efflux from normal human skin fibroblasts but not from fibroblasts from a Tangier patient . Fluorescent confocal microscopy and biotinylation assay demonstrated that in J774 cells probucol impaired the translocation of O95477 from intracellular compartments to the plasma membrane . DB01599 also inhibited the formation of an O95477 -linked cholesterol oxidase sensitive plasma membrane domain . Consistent with the inhibitory effect on O95477 translocation to the plasma membrane , probucol reduced cell surface-specific [ 125I ] -labeled apolipoprotein-AI binding . CONCLUSIONS : We conclude that probucol is an effective inhibitor of O95477 -mediated cholesterol efflux without influencing scavenger receptor class B type I-mediated efflux . The inhibition of O95477 translocation to the plasma membrane may in part explain the reported in vivo high-density lipoprotein-lowering action of probucol . Q14954 -positive NK cells mediate alloresponse against the P06681 HLA- P55040 ligand group in vitro . The inhibitory 2DL1 and activating 2DS1 killer Ig-like receptors ( P55040 ) both have shared ligand specificity for codon sequences in the P06681 group HLA-Cw Ags . In this study , we have investigated NK cell activation by allogeneic target cells expressing different combinations of the HLA- P55040 ligand groups C1 , P06681 , and Bw4 . We demonstrate that fresh NK cells as well as P60568 -propagated NK cells from 2DS1-positive donors that are homozygous for the C1 ligand group are activated in vitro by B lymphoblastoid cell lines expressing the P06681 group . This response is , in part , due to the absence of C1 group recognition mediated by the inhibitory receptor 2DL2/3 . This " missing self " alloresponse to P06681 , however , is rarely observed in NK cells from donors lacking 2DS1 . Even in presence of 2DS1 , the NK alloresponse is dramatically reduced in donors that have P06681 group as " self. " Analysis of selected NK clones that express 2DS1 mRNA and lack mRNA for 2DL1 demonstrates that activation by the P06681 ligand and mAb cross-linking of 2DS1 in these clones induces P01579 . Furthermore , this P06681 group-induced activation is inhibited by Abs to both HLA class I and the receptor . Collectively , these studies demonstrate that NK cells from 2DS1-positive donors are activated by target cells that express the P06681 group as an alloantigen . This leads to increased P01579 -positive fresh NK cells and induces NK allocytotoxicity in P60568 -propagated polyclonal NK cells and NK clones . This study also provides support for the concept that incompatibility for the HLA- P55040 ligand groups C1 , P06681 , and Bw4 dominates NK alloactivation in vitro . Prevention of acute and chronic allograft rejection by a novel retinoic acid receptor-alpha-selective agonist . To investigate the involvement of retinoic acid receptor ( RAR ) -alpha in allograft rejection , we investigated the effect of a novel selective agonist to the receptor , ER-38925 , in a mouse cardiac allograft model . Prophylactic treatment with ER-38925 inhibited the acute rejection of the mouse cardiac allograft ( BALB/c --> C3H/HeN ) at 0.3 and 3 mg/kg , and its effect was enhanced in combination with tacrolimus . In this model , ER-38925 remarkably inhibited cytotoxic T lymphocyte induction and alloantigen-stimulated production of cytokines , i.e. P60568 , IL-12 and P01579 . In the chronic rejection model , combined treatment with tacrolimus and ER-38925 reduced the grade and incidence of arteriosclerosis in the cardiac allografts significantly more potently than tacrolimus monotherapy . ER-38925 inhibited the proliferation of rat aortic smooth muscle cells stimulated in vitro , presumably through the induction of a cyclin-dependent kinase inhibitor , p27(kip-1) . Those results provide a rationale for using P10276 agonists as immunosuppressants in human organ transplantation . Fluorescence energy transfer analysis of calmodulin-peptide complexes . The interactions between calmodulin and the tryptophan residues of synthetic peptides corresponding to the calmodulin binding domains of skeletal muscle myosin light-chain kinase and the plasma membrane calcium pump were examined . The single tryptophan residue contained in each peptide became relatively immobilized and inaccessible to iodide ion upon binding to calmodulin , indicating that the indole side chain was inserted into a hydrophobic cleft in the surface of calmodulin . Fluorescence energy transfer from peptidyl tryptophan residues to an AEDANS moiety attached to cysteine-26 of spinach calmodulin was measured . Included in these analyses was a tryptophan-containing peptide analog of the calmodulin binding domain of neuromodulin . These data indicated that the indole ring of each peptide inserted 32-35 A away from cysteine-26 and may therefore interact with the carboxyl-terminal lobe of P62158 in its " bent " conformation [ Persechini & Kretsinger ( 1988a ) J. Cardiovasc. Pharmacol. 12 ( Suppl 5 ) , S1- P28222 ; Ikura et al. ( 1992 ) Science 256 , 632-638 ; Vorherr et al. ( 1992 ) Eur. J. Biochem. 204 , 931-937 ] . The interchange of tryptophan-3 and phenylalanine-21 of the calcium pump peptide increased the efficiency of energy transfer to the AEDANS-moiety approximately 12-fold , reducing the calculated distance to 20 A . These data suggest that phenylalanine-21 of the calcium pump peptide interacts with the hydrophobic cleft in the amino-terminal lobe of P62158 . Immunomodulatory role of high-density lipoproteins : impact on immunosenescence . Natural aging is accompanied by a dysregulation of the host immune response that has well-known clinical consequences but poorly defined underlying causes . It has previously been reported that advancing age is associated with an increase in membrane cholesterol level in T cells . The aim of this study was to investigate whether high-density lipoprotein ( HDL ) can modulate the age-related accumulation of membrane cholesterol in T cells and impact on their subsequent responsiveness . Our data reveal that cholesterol metabolism , influx , and efflux are altered in T cells with aging , which may in part explain the increase in membrane cholesterol level observed in T cells in elderly individuals . HDL was unable to promote reverse cholesterol transport in T cells from elderly subjects with the same efficiency as was observed in T cells from young subjects besides unchanged ABCA-1 and Q8WTV0 expressions . HDL exhibited a short-acting co-stimulatory effect by enhancing T cell production of interleukin-2 ( P60568 ) . Moreover , HDL from healthy normolipemic individuals exerted differential effects on T cell proliferation that depended on the age of the HDL donor . Finally , HDL modulated TCR/ P10747 activation by inducing sustained signaling through pLck , pERK , and pAkt . These data suggest that HDL has immunomodulatory effects on T cells that are influenced by age . Regulation of porphyrin synthesis and photodynamic therapy in heavy metal intoxication . Protoporphyrin IX ( PpIX ) synthesis by malignant cells is successfully exploited for photodynamic therapy ( PDT ) following administration of 5-aminolevulinic acid ( ALA ) and light irradiation . The influence of two environmental heavy metal poisons , lead and gallium , on PpIX-synthesis and DB00855 was studied in two neu-ronal cell lines , SH-SY5Y neuroblastoma and PC12 pheochromocytoma . The heavy metal intoxication affected two of the heme-synthesis enzymes , ALA-dehydratase ( P13716 ) and porphobilinogen deaminase ( P08397 ) . The present results show that lead poisoning significantly decreased the P08397 cellular level and inhibited its enzymatic activity , whereas the effects of gallium were less prominent . Although , the protein levels were reduced , the mRNA levels of P08397 remained unchanged during metal intoxication . These findings show additional inhibitory activity of lead on top of its classical effect on P13716 . Proteasome activity was enhanced during lead treatment , as measured by the AMC fluorigenic proteasome assay . The reduction in P08397 levels was not a consequence of P08397 mRNA reduced synthesis , which remained unchanged as shown by RT-PCR analysis . As a result of the lead poisoning , marked alterations in the cell cycle were observed , including a decreased P55008 phase and an increased number of S phase cells . The efficacy of DB00855 was reduced in correlation with decreased activities of the enzymes during lead intoxication . We may conclude that lead poisoning adversely affects the outcome of ALA photodynamic therapy of cancer . Chronic methamphetamine exposure alters immune function in normal and retrovirus-infected mice . Methamphetamine ( MA ) abuse represents a growing problem in the USA with an increase of sudden death . To evaluate the immune function alterations due to chronic methamphetamine use , we examined C57BL/C mice with LP-BM5 retrovirus infection plus methamphetamine exposure . Mice were randomly assigned to the following groups : placebo , placebo retrovirus-infected , uninfected MA treated and retrovirus-infected MA treated . Placebo , MA-treated groups were intraperitoneally injected with saline , MA , respectively , with a gradually increasing dose from 15 to 40 mg/kg for 12 weeks ( 5 days/week ) . Con A- and LPS-induced mitogenesis of splenocytes , cytokine production by splenocytes culture and lipid peroxides in the liver were measured . Heart tissue histopathology was analyzed in all the groups with murine cytomegalovirus ( CMV ) superinfection . Our data showed that MA treatment significantly decreased production of P60568 and interferon gamma ( P01579 ) in uninfected mice but did not further suppress the reduced Th1 cytokines in retrovirus-infected mice . There were no significant effects on cytokines P05112 and P05231 . However , tumor necrosis factor ( P01375 ) was significantly increased in both uninfected and infected mice due to MA treatment . Lipid peroxides in liver were significantly increased both in uninfected and retrovirus-infected mice due to MA exposure . DB00163 levels in liver were significantly decreased in uninfected mice due to MA treatment . CMV superinfection greatly increased the cardiac lesions in retrovirus-infected mice while no significant histopathology changes were detected due to MA treatment . Our data suggest that MA has immunomodulation activity , suppressing Th1 cytokine production and enhancing some Th2 cytokine secretion , as well as increasing lipid peroxides in uninfected mice . The interaction between LP-BM5 and MA remains unclear . Postnatal development of serotonin 1B , 2 A and 2C receptors in brainstem motoneurons . The effects of serotonin ( 5-HT ) on motoneurons are mediated via multiple receptor subtypes . In hypoglossal ( XII ) motoneurons , the prototypic brainstem motoneurons whose functions change during the postnatal period , 5-HT effects evolve from inhibitory to excitatory , probably in association with changes in receptor expression . We studied P28222 , 5- Q13049 and P28335 receptor mRNA in 414 dissociated XII motoneurons and 5- Q13049 protein in the XII , facial and spinal cervical ( P06681 -3 ) motor nuclei . The percentage of motoneurons expressing distinct mRNAs varied with the postnatal age ( P09131 -33 days ) and receptor subtype . Initially , P28222 mRNA was present in 50-85 % of cells , but on P14 its expression transiently decreased below 35 % . 5- Q13049 mRNA was present in nearly all cells after P6 , but in less than 65 % on P09131 -5 . Normal and/or short splice variants of the P28335 mRNA were expressed in less than 20 % of motoneurons on P09131 -9 , and in approximately 35 % thereafter . P28222 and 5- Q13049 mRNAs often were expressed in different cells during early and intermediate postnatal periods , whereas P28335 mRNA never occurred alone . The 5- Q13049 receptor protein level gradually increased through P15 in the XII and facial nuclei , with dendritic labelling appearing in XII motoneurons only after P12 . In spinal motoneurons , both somatic and dendritic labelling was strongest on Q15084 and then decreased . The development of 5-HT receptors in XII motoneurons may be related to changes in feeding behaviour , whereas different cues regulate 5-HT receptor expression in upper spinal motoneurons . A Minireview on DB00067 -regulated P41181 in Kidney Collecting Duct Cells . The kidney collecting duct is an important renal tubular segment for the regulation of body water and salt homeostasis . DB09145 reabsorption in the collecting duct cells is regulated by arginine vasopressin ( AVP ) via the vasopressin V2-receptor ( P30518 ) . AVP increases the osmotic water permeability of the collecting duct cells through aquaporin-2 ( P41181 ) and aquaporin-3 ( Q92482 ) . AVP induces the apical targeting of P41181 and transcription of P41181 gene in the kidney collecting duct principal cells . The signaling transduction pathways resulting in the P41181 trafficking to the apical plasma membrane of the collecting duct principal cells , include P41181 phosphorylation , RhoA phosphorylation , actin depolymerization and calcium mobilization , and the changes of P41181 protein abundance in water balance disorders have been extensively studied . These studies elucidate the underlying cellular and molecular mechanisms of body water homeostasis and provide the basis for the treatment of body water balance disorders . Heart allograft protection with low-dose carbon monoxide inhalation : effects on inflammatory mediators and alloreactive T-cell responses . BACKGROUND : DB11588 ( CO ) , a byproduct of heme catalysis , has lately received considerable attention as a regulatory molecule in cellular and biological processes . CO has been shown to provide potent protection against a variety of tissue injuries . We hypothesized in this study that low concentration CO would be beneficial for organ allografts , which frequently undergo several types of injury such as ischemia/reperfusion , alloimmune reaction , and inflammation METHODS : The efficacy of low-dose CO was examined in a fully allogeneic LEW to BN rat heterotopic heart transplantation ( HHTx ) model . Recipients were kept in air or exposed to low-dose CO ( 20 ppm ) for 14 , 28 , or 100 days after HHTx under short-course tacrolimus RESULTS : CO treatment ( d0-28 , 0-100 ) was remarkably effective in prolonging heart allograft survival to a median of > 100 from 45 days in the air-control group , with significant reductions of arteritis , fibrosis , and cellular infiltration , including macrophages and T cells . CO inhibited intragraft upregulation of Th1 type cytokines ( P60568 , IFNgamma ) , proinflammatory mediators ( IL-1beta , TNFalpha , P05231 , P35354 ) , and adhesion molecule . Shorter CO exposure in early ( 0-13d ) and late ( 14-28d ) posttransplant periods also prolonged graft survival , with a significant inhibition of inflammatory mediators CONCLUSIONS : These results show that low dose CO inhalation protects heart allografts and can considerably prolong their survival . CO appears to function via multiple mechanisms , including direct inhibition of Th1 type cytokine production and regulation of inflammatory responses . Farnesyl diphosphate synthase : the art of compromise between substrate selectivity and stereoselectivity . Farnesyl diphosphate ( FPP ) synthase catalyzes the consecutive head-to-tail condensations of isopentenyl diphosphate ( IPP , P01031 ) with dimethylallyl diphosphate ( DMAPP , P01031 ) and geranyl diphosphate ( GPP , Q99622 ) to give ( E,E ) -FPP ( C15 ) . The enzyme belongs to a genetically distinct family of chain elongation enzymes that install E-double bonds during each addition of a five-carbon isoprene unit . Analysis of the Q99622 and C15 products from incubations with avian P14324 reveals that small amounts of neryl diphosphate ( Z- Q99622 ) and ( Z,E ) -FPP are formed along with the E-isomers during the P01031 --> Q99622 and Q99622 --> C15 reactions . Similar results were obtained for P14324 from Escherichia coli , Artemisia tridentata ( sage brush ) , Pyrococcus furiosus , and Methanobacter thermautotrophicus and for GPP and FPP synthesized in vivo by E. coli P14324 . When ( R ) -[2-2H]IPP was a substrate for chain elongation , no deuterium was found in the chain elongation products . In contrast , the deuterium in ( S ) -[2-2H]IPP was incorporated into all of the products . Thus , the pro-R hydrogen at P06681 of IPP is lost when the E- and Z-double bond isomers are formed . The synthesis of Z-double bond isomers by P14324 during chain elongation is unexpected for a highly evolved enzyme and probably reflects a compromise between optimizing double bond stereoselectivity and the need to exclude DMAPP from the IPP binding site . In vivo effects of a combined P28222 receptor/ P31645 antagonist in experimental pulmonary hypertension . AIMS : A mechanism for co-operation between the serotonin ( 5-hydroxytryptamine , 5-HT ) transporter and P28222 receptor in mediating pulmonary artery vasoconstriction and proliferation of pulmonary artery smooth muscle cells has been demonstrated in vitro . Here we determine , for the first time , the in vivo effects of a combined P28222 receptor/serotonin transporter antagonist ( LY393558 ) with respect to the development of pulmonary arterial hypertension ( PAH ) and its in vitro effects in human pulmonary artery smooth muscle cells ( hPASMCs ) derived from idiopathic PAH ( IPAH ) patients . METHODS AND RESULTS : We determined the effects of LY393558 as well as a selective serotonin transporter inhibitor , citalopram , on right ventricular pressure , right ventricular hypertrophy , and pulmonary vascular remodelling in wildtype mice and mice over-expressing serotonin transporter ( P31645 + mice ) before and after hypoxic exposure . We also compared their effectiveness at reversing PAH in P31645 + mice and hypoxic mice . Further , we examined the proliferative response to serotonin in IPAH hPASMCs . We also clarified the pharmacology of serotonin-induced vasoconstriction and P28222 receptor/serotonin transporter interactions in mouse isolated pulmonary artery . DB00215 had a moderate effect at preventing and reversing experimental PAH in vivo whereas LY393558 was more effective . LY393558 was more effective than citalopram at reversing serotonin-induced proliferation in IPAH hPASMCs . There is synergy between P28222 receptor and serotonin transporter inhibitors against serotonin-induced vasoconstriction in mouse pulmonary arteries . CONCLUSION : P28222 receptor and serotonin transporter inhibition are effective at preventing and reversing experimental PAH and serotonin-induced proliferation of PASMCs derived from IPAH patients . Targeting both the serotonin transporter and P28222 receptor may be a novel therapeutic approach to PAH . In vitro cell death of activated lymphocytes in Omenn 's syndrome . Omenn 's syndrome ( OS ) is characterized by erythrodermia , hepatosplenomegaly , lymphadenopathy , hypereosinophilia and elevated IgE levels associated with increased susceptibility to severe infections . Peripheral blood T cells , though usually present in normal number , show an activated phenotype ( including an increased expression of CD95/Fas ) , a Th2 pattern of cytokine secretion and defective proliferative response to mitogens . In this report , we demonstrate that T cells from patients with OS undergo an excessive cell death in vitro resulting from two mechanisms . First , a substantial number of peripheral blood lymphocytes from OS patients die in unstimulated cultures ( p = 0.009 vs. healthy controls ) . This spontaneous apoptosis is associated with reduced expression of bcl-2 gene product ( p < 0.05 ) and can be prevented by addition of interleukin ( IL ) -2 ( which also prevents down-modulation of bcl-2 ) , while is independent from CD95 signaling . Second , lymphocytes from OS patients are highly susceptible to activation-induced cell death ( AICD ) induced with mitogens . This mechanism is largely independent from P60568 , while it can be significantly inhibited blocking CD95 with an IgG2b monoclonal antibody ( mAb ) . The dependence of AICD from signals transduced via CD95 was confirmed showing that cross-linking CD95 with an IgM mAb induces a higher cell death in purified P01730 + CD45R0+ cells from OS patients than in controls ( comparable for CD95 expression ) . Both mechanisms of cell death observed in this study result from lymphocyte hyperactivation occurring in vivo in these patients and may contribute to functional T cell defects of OS . The cholinergic system is involved in regulation of the development of the hematopoietic system . Gene expression profiling demonstrated that components of the cholinergic system , including choline acetyltransferase , acetylcholinesterase and nicotinic acetylcholine receptors ( nAChRs ) , are expressed in embryonic stem cells and differentiating embryoid bodies ( EBs ) . Triggering of nAChRs expressed in EBs by nicotine resulted in activation of MAPK and shifts of spontaneous differentiation toward hemangioblast . In vivo , non-neural nAChRs are detected early during development in fetal sites of hematopoiesis . Similarly , in vivo exposure of the developing embryo to nicotine resulted in higher numbers of hematopoietic progenitors in fetal liver . However postpartum , the number of hematopoietic stem/progenitor cells ( O14818 ) was decreased , suggesting an impaired colonization of the fetal bone marrow with HSPCs . This correlated with increased number of circulating O14818 and decreased expression of P61073 that mediates migration of circulating cells into the bone marrow regulatory niche . In addition , protein microarrays demonstrated that nicotine changed the profile of cytokines produced in the niche . While the levels of IL1alpha , IL1beta , P60568 , P15248 and P22301 were not changed , the production of hematopoiesis-supportive cytokines including DB00099 , GM- P04141 , P08700 , P05231 and P17936 was decreased . This correlated with the decreased repopulating ability of O14818 in vivo and diminished hematopoietic activity in bone marrow cultures treated with nicotine . Interestingly , nicotine stimulated the production of P05112 and P05113 , implying a possible role of the cholinergic system in pathogenesis of allergic diseases . Our data provide evidence that the nicotine-induced imbalance of the cholinergic system during gestation interferes with normal development and provides the basis for negative health outcomes postpartum in active and passive smokers . N6-isopentenyladenosine , an endogenous isoprenoid end product , directly affects cytotoxic and regulatory functions of human NK cells through P14324 modulation . iPA is a naturally occurring nucleoside with an isopentenyl moiety derived from the mevalonate pathway and a well-established anti-tumor activity . In analogy to the unique specificity for phosphoantigens , such as IPP , shown by human Vγ9Vδ2 T cells , here , we report for the first time the ability of iPA to selectively expand and directly target human NK cells . Interestingly , submicromolar doses of iPA stimulate resting human NK cells and synergize with P60568 to induce a robust activation ex vivo with significant secretion of P13501 and P10147 and a large increase in P01375 -α and IFN-γ production when compared with P60568 single cytokine treatment . Moreover , iPA promotes NK cell proliferation and up-regulates the expression of specific NK cell-activating receptors , as well as Q07108 and CD107a expression . Accordingly , this phenotype correlates with significantly greater cytotoxicity against tumor targets . At the molecular level , iPA leads to a selective , potent activation of MAPK signaling intermediaries downstream of the IL-2R . The effect results , at least in part , from the fine modulation of the P14324 activity , the same enzyme implicated in the stimulation of the human γδ T cells . The iPA-driven modulation of P14324 can cause an enhancement of post-translational prenylation essential for the biological activity of key proteins in NK signaling and effector functions , such as Ras . These unanticipated properties of iPA provide an additional piece of evidence of the immunoregulatory role of the intermediates of the mevalonate pathway and open novel therapeutic perspectives for this molecule as an immune-modulatory drug . Gene transcription abnormalities in canine atopic dermatitis and related human eosinophilic allergic diseases . Canine atopic dermatitis ( AD ) is clinically similar to human AD , implicating it as a useful model of human eosinophilic allergic disease . To identify cutaneous gene transcription changes in relatively early inflammation of canine AD , microarrays were used to monitor transcription in normal skin ( n=13 ) and in acute lesional AD ( P13716 ) and nearby visibly nonlesional AD ( NLAD ) skin ( n=13 ) from dogs . Scanning the putative abnormally transcribed genes , several potentially relevant genes , some abnormally transcribed in both NLAD and P13716 ( e.g. P05231 , Q8NET5 , Q9UJ68 , and P43405 ) , were observed . Comparison for abnormally transcribed genes common to two related human diseases , human AD and asthmatic chronic rhinosinusitis with nasal polyps ( aCRSwNP ) , further identified genes or gene sets likely relevant to eosinophilic allergic inflammation . These included : ( 1 ) genes associated with alternatively activated monocyte-derived cells , including members of the monocyte chemotactic protein ( MCP ) gene cluster , ( 2 ) members of the IL1 family gene cluster , ( 3 ) eosinophil-associated seven transmembrane receptor Q14246 and Q9BY15 genes , ( 4 ) interferon-inducible genes , and ( 5 ) keratin genes associated with hair and nail formation . Overall , numerous abnormally transcribed genes were observed only in canine AD ; however , many others are common to related human eosinophilic allergic diseases and represent therapeutic targets testable in dogs with AD . Effect of axitinib ( AG-013736 ) on fatigue , thyroid-stimulating hormone , and biomarkers : a phase I study in Japanese patients . DB06626 is an oral , potent , and selective inhibitor of vascular endothelial growth factor receptor ( VEGFR ) 1 , 2 , and 3 . This phase I study evaluated the safety , pharmacokinetics , pharmacodynamics , antitumor activity , and recommended starting dose of axitinib in patients with advanced solid tumors . Twelve patients received single-dose axitinib 5 mg and were monitored for > or =48 h . Continuous 5 mg twice-daily dosing was then initiated . One patient had dose-limiting toxicity ( grade 3 proteinuria and fatigue ) . Common treatment-related adverse events were anorexia , fatigue , and diarrhea . Grade 3 treatment-related adverse events were fatigue and hypertension . Maximum axitinib plasma concentration occurred 1-4 h after steady-state dosing . Eleven patients experienced thyroid-stimulating hormone elevation ; time-course change and fatigue onset appeared to be related in some patients . Significant correlation was observed between thyroid-stimulating hormone change and area under the plasma concentration-time curve ( AUC ; r = 0.80 , P = 0.005 ) . DB06626 decreased plasma soluble vascular endothelial growth factor receptor 2 ( s- P35968 ) , with significant correlation between change in s- P35968 and AUC ( r = -0.92 , P < 0.0001 ) . Fluorodeoxyglucose positron emission tomography revealed a substantial decrease in tumor metabolic activity associated with axitinib . Tumor size decreased in nine patients . The time-course of thyroid-stimulating hormone change appeared correlated with fatigue . There were significant correlations between thyroid-stimulating hormone or s- P35968 and axitinib exposure . DB06626 5 mg twice-daily is the recommended starting dose for Japanese patients . This trial is registered with ClinicalTrials.gov , identifier NCT00447005 . The Retinoic Acid Receptor-alpha mediates human T-cell activation and Th2 cytokine and chemokine production . BACKGROUND : We have recently demonstrated that all-trans-retinoic acid ( DB00755 ) and 9-cis-retinoic acid ( 9-cis RA ) promote P05112 , P05113 and P35225 synthesis , while decreasing P01579 and P01375 expression by activated human T cells and reduces the synthesis of IL-12p70 from accessory cells . Here , we have demonstrated that the observed effects using DB00755 and 9-cis RA are shared with the clinically useful RAR ligand , 13-cis retinoic acid ( 13-cis RA ) , and the retinoic acid receptor-alpha ( P10276 ) -selective agonist , AM580 but not with the P10826 /gamma ligand , 4-hydroxyphenylretinamide ( DB05076 ) . RESULTS : The increase in type 2 cytokine production by these retinoids correlated with the expression of the T cell activation markers , Q07108 and P28907 . The P10276 -selective agonist , AM580 recapitulated all of the T cell activation and type 2 cytokine-inducing effects of DB00755 and 9- DB00982 , while the P10276 -selective antagonist , RO 41-5253 , inhibited these effects . CONCLUSION : These results strongly support a role for P10276 engagement in the regulation of genes and proteins involved with human T cell activation and type 2 cytokine production .