MH_dev_236

Safety , pharmacokinetics and pharmacodynamics of four-hour intravenous infusions of eritoran in healthy Japanese and Caucasian men . DB04933 MEN , a synthetic analogue of lipid A , has been shown to bind to O00206 REA / MD - 2 complex and thereby block the interaction of endotoxins with O00206 REA . We report here the results of a study conducted to assess the single-dose safety and tolerability , as well as the pharmacokinetics and pharmacodynamics , of eritoran infusion in Japanese and Caucasian healthy adult men . Sixty-four men ( aged 20-45 years ; body mass index 18-30 kg / m ( 2 ) ) were randomized into four groups : 4 - mg total dose ( six Japanese and six Caucasian men ) ; 12 - mg total dose ( 12 Japanese and 12 Caucasian men ) ; 28 - mg total dose ( six Japanese and six Caucasian men ) ; and placebo ( eight Japanese and eight Caucasian men ) . DB04933 MEN in single doses up to 28 mg over 4 h was well tolerated , with no apparent ethnic differences noted . Plasma concentrations were slightly higher in Japanese versus Caucasian men ; these differences were not significant after adjustment for differences in body mass ( clearance : approximately 1.2 ml / h / kg ; volume of distribution at steady state : approximately 0.07 l / kg ) . The ex vivo endotoxin inhibitory activity of eritoran was similar in Japanese and Caucasian men . The data do not indicate any need for clinical dose adjustment for possible ethnic-based differences in drug distribution or metabolism .

Initial testing of the multitargeted kinase inhibitor pazopanib by the Pediatric Preclinical Testing Program . DB06589 SUB is an oral angiogenesis inhibitor targeting vascular growth factor receptor - 1 , - 2 , and - 3 , platelet derived growth factor receptor-α , platelet derived growth factor receptor-β , and P10721 REA that has demonstrated activity against a variety of adult cancer xenografts . DB06589 SUB was tested against a panel of pediatric rhabdomyosarcoma and Ewing sarcoma xenografts at a dose of 108 mg / kg / day or 100 mg / kg twice daily , administered orally for 28 days . While no objective responses were observed , pazopanib induced statistically significant differences in event-free survival compared to controls in approximately one-half of the sarcoma xenograft models tested . Though well tolerated , pazopanib showed limited activity against the sarcoma models evaluated , with the best tumor responses being growth delay .

Effective antibody therapy induces host-protective antitumor immunity that is augmented by O00206 REA agonist treatment . Toll-like receptors are potent activators of the innate immune system and generate signals leading to the initiation of the adaptive immune response that can be utilized for therapeutic purposes . We tested the hypothesis that combined treatment with a Toll-like receptor agonist and an antitumor monoclonal antibody is effective and induces host-protective antitumor immunity . C57BL / 6 human mutated P04626 REA ( hmHER 2 ) transgenic mice that constitutively express kinase-deficient human P04626 REA under control of the CMV promoter were established . These mice demonstrate immunological tolerance to D5 - P04626 REA , a syngeneic human P04626 REA - expressing melanoma cell line . This human P04626 REA - tolerant model offers the potential to serve as a preclinical model to test both antibody therapy and the immunization potential of human P04626 REA - targeted therapeutics . Here , we show that E6020 , a Toll-like receptor - 4 ( O00206 REA ) agonist effectively boosted the antitumor efficacy of the monoclonal antibody trastuzumab in immunodeficient C57BL / 6 SCID mice as well as in C57BL / 6 hmHER 2 transgenic mice . E6020 and trastuzumab co-treatment resulted in significantly greater inhibition of tumor growth than was observed with either agent individually . Furthermore , mice treated with the combination of trastuzumab and the O00206 REA agonist were protected against rechallenge with human P04626 REA - transfected tumor cells in hmHER 2 transgenic mouse strains . These findings suggest that combined treatment with trastuzumab and a O00206 REA agonist not only promotes direct antitumor effects but also induces a host-protective human P04626 REA - directed adaptive immune response , indicative of a memory response . These data provide an immunological rationale for testing O00206 REA agonists in combination with antibody therapy in patients with cancer .

P15121 REA inhibition suppresses azoxymethane-induced colonic premalignant lesions in C57BL / KsJ-db / db mice . Type - 2 diabetes and obesity-related metabolic abnormalities are major risk factors for the development of colon cancer . In the present study , we examined the effects of polyol pathway enzyme aldose reductase ( AR ) inhibitor , fidarestat , on the development of azoxymethane ( AOM ) - induced colonic premalignant lesions in C57BL / KsJ-db / db obese mice . Our results indicate that fidarestat given in the drinking water caused a significant reduction in the total number of colonic premalignant lesions in the AOM treated obese mice . Further , the expression levels of PKC-β 2 , AKT , P35354 REA and P35228 REA in the colonic mucosa of AOM-treated mice were significantly decreased by fidarestat . The serum levels of IL - 1α , P02778 REA , Q07325 REA , P01375 REA - α and P15692 REA are significantly suppressed in AOM + fidarestat treated obese mice . DB02021 MEN also decreased the expression of P35354 REA , P35228 REA , P98170 REA , survivin , β-catenin and NF-κB in high glucose-treated HT29 colon cancer cells . In conclusion , our results indicate that fidarestat inhibits the development of colonic premalignant lesions in an obesity-related colon cancer and is chemopreventive to colorectal carcinogenesis in obese individuals .

P35354 REA predicts adverse effects of tamoxifen : a possible mechanism of role for nuclear P04626 REA in breast cancer patients . P35354 REA ( P35354 REA ) is associated with breast tumour progression . Clinical and molecular studies implicate human epidermal growth factor receptor 2 ( P04626 REA ) in the regulation of P35354 REA expression . Recent reports raise the possibility that P04626 REA could mediate these effects through direct transcriptional mechanisms . The relationship between P04626 REA and P35354 REA was investigated in a cohort of breast cancer patients with or without endocrine treatment . A tissue microarray comprising tumours from 560 patients with 10 - year follow-up was analysed for P04626 REA , P27361 REA / 2 , polyoma enhancer activator 3 ( P43268 REA ) and P35354 REA expression . Subcellular localisation of P04626 REA was assessed by immunofluorescence and confocal microscopy . Expression of markers examined was analysed in relation to classic clinicopathological parameters and disease-free survival in the presence and absence of tamoxifen . P35354 REA expression associated with both membranous and nuclear expression of P04626 REA ( P= 0.0033 and P < 0.00001 respectively ) . No association was detected between P35354 REA and either P27361 REA / 2 or P43268 REA ( P= 0.7 and P= 0.3 respectively ) . None of the markers were found to be independently prognostic . Membrane P04626 REA , nuclear P04626 REA and P35354 REA , however , were all found to predict poor disease-free survival in patients on endocrine treatment ( P= 0.0017 , P= 0.0003 and P= 0.0202 respectively ) . Moreover , patients who were positive for P35354 REA predicted adverse effects of tamoxifen ( P= 0.0427 ) . These clinical ex vivo data are consistent with molecular observations that P04626 REA can regulate P35354 REA expression through direct transcriptional mechanisms . P35354 REA expression correlates with disease progression on endocrine treatment . This study supports a role for P35354 REA as a predictor of adverse effects of tamoxifen in breast cancer patients .

Relatively selective neuronal nitric oxide synthase inhibition by 7 - nitroindazole in monkey isolated cerebral arteries . The selectivity of 7 - nitroindazole in inhibiting endothelial and neuronal nitric oxide synthases ( P29474 REA and P29475 REA ) was investigated by comparing its inhibitory action on relaxations mediated by nitric oxide ( NO ) in response to stimulation of perivascular nerves and in response to histamine in monkey cerebral artery strips . DB02207 MEN at 2 x 10 ( - 5 ) M moderately attenuated the response to transmural electrical stimulation and to nicotine , but did to alter the endothelium-dependent relaxation in response to histamine in cimetidine-treated strips . Raising the concentration of 7 - nitroindazole to 10 ( - 4 ) M abolished the neurogenic response , partially inhibited the histamine-induced relaxation , but did not affect the response to NO . It is concluded that 7 - nitroindazole is a relatively selective P29475 REA inhibitor ; however , at high concentrations , it inhibits P29474 REA in monkey cerebral arteries .

Neovastat ( DB05387 MEN ) inhibits the airway inflammation and hyperresponsiveness in a murine model of asthma . Matrix metalloproteinase ( MMP ) - 9 plays an important role in the pathogenesis of bronchial asthma . Neovastat , having significant antitumor and antimetastatic properties , is classified as a naturally occurring multifunctional antiangiogenic agent . We evaluated the therapeutic effect of Neovastat on airway inflammation in a mouse model of asthma . BALB / c mice were immunized subcutaneously with ovalbumin ( OVA ) on days 0 , 7 , 14 , and 21 and challenged with inhaled OVA on days 26 , 29 , and 31 . Neovastat was administrated by gavage ( 5 mg / kg body weight ) three times with 12 h intervals , beginning 30 min before OVA inhalation . On day 32 , mice were challenged with inhaled methacholine , and enhanced pause ( Penh ) was measured as an index of airway hyperresponsiveness . The severity of airway inflammation was determined by differential cell count of bronchoalveolar lavage ( BAL ) fluid . The P14780 REA concentration in BAL fluid samples was measured by ELISA , and P14780 REA activity was measured by zymography . The untreated asthma group showed an increased inflammatory cell count in BAL fluid and Penh value compared with the normal control group . Mice treated with Neovastat had significantly reduced Penh values and inflammatory cell counts in BAL fluid compared with untreated asthmatic mice . Furthermore , mice treated with Neovastat showed significantly reduced P14780 REA concentrations and activity in BAL fluid . These results demonstrate that Neovastat might have new therapeutic potential for airway asthmatic inflammation .

DB08896 MEN ( DB08896 MEN ) : a new oral multikinase inhibitor of angiogenic , stromal and oncogenic receptor tyrosine kinases with potent preclinical antitumor activity . Angiogenesis , a critical driver of tumor development , is controlled by interconnected signaling pathways . Vascular endothelial growth factor receptor ( VEGFR ) 2 and tyrosine kinase with immunoglobulin and epidermal growth factor homology domain 2 play crucial roles in the biology of normal and tumor vasculature . DB08896 MEN ( DB08896 MEN ) , a novel oral multikinase inhibitor , potently inhibits these endothelial cell kinases in biochemical and cellular kinase phosphorylation assays . Furthermore , regorafenib inhibits additional angiogenic kinases ( P17948 REA / 3 , platelet-derived growth factor receptor-β and fibroblast growth factor receptor 1 ) and the mutant oncogenic kinases P10721 REA , P07949 REA and B-RAF . The antiangiogenic effect of regorafenib was demonstrated in vivo by dynamic contrast-enhanced magnetic resonance imaging . DB08896 MEN administered once orally at 10 mg / kg significantly decreased the extravasation of Gadomer in the vasculature of rat GS9L glioblastoma tumor xenografts . In a daily ( qd ) × 4 dosing study , the pharmacodynamic effects persisted for 48 hr after the last dosing and correlated with tumor growth inhibition ( TGI ) . A significant reduction in tumor microvessel area was observed in a human colorectal xenograft after qd × 5 dosing at 10 and 30 mg / kg . DB08896 MEN exhibited potent dose-dependent TGI in various preclinical human xenograft models in mice , with tumor shrinkages observed in breast MDA-MB - 231 and renal 786 - O carcinoma models . Pharmacodynamic analyses of the breast model revealed strong reduction in staining of proliferation marker Ki - 67 and phosphorylated extracellular regulated kinases 1/2 . These data demonstrate that regorafenib is a well-tolerated , orally active multikinase inhibitor with a distinct target profile that may have therapeutic benefit in human malignancies .

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 .

The postnatal rat aorta contains pericyte progenitor cells that form spheroidal colonies in suspension culture . Pericytes play an important role in modulating angiogenesis , but the origin of these cells is poorly understood . To evaluate whether the mature vessel wall contains pericyte progenitor cells , nonendothelial mesenchymal cells isolated from the rat aorta were cultured in a serum-free medium optimized for stem cells . This method led to the isolation of anchorage-independent cells that proliferated slowly in suspension , forming spheroidal colonies . This process required basic fibroblast growth factor ( P09038 REA ) in the culture medium , because P09038 REA withdrawal caused the cells to attach to the culture dish and irreversibly lose their capacity to grow in suspension . Immunocytochemistry and RT-PCR analysis revealed the expression of the precursor cell markers P28906 REA and Tie - 2 and the absence of endothelial cell markers ( CD31 and endothelial nitric oxide synthase , P29474 REA ) and smooth muscle cell markers ( alpha-smooth muscle actin , alpha-SMA ) . In addition , spheroid-forming cells were positive for Q99942 REA , nestin , PDGF receptor ( P09619 REA ) - alpha , and P09619 REA . Upon exposure to serum , these cells lost P28906 REA expression , acquired alpha-SMA , and attached to the culture dish . Returning these cells to serum-free medium failed to restore their original spheroid phenotype , suggesting terminal differentiation . When embedded in collagen gels , spheroid-forming cells rapidly migrated in response to DB00102 and became dendritic . Spheroid-forming cells cocultured in collagen with angiogenic outgrowths of rat aorta or isolated endothelial cells transformed into pericytes . These results demonstrate that the rat aorta contains primitive mesenchymal cells capable of pericyte differentiation . These immature cells may represent an important source of pericytes during angiogenesis in physiological and pathological processes . They may also provide a convenient supply of mural cells for vascular bioengineering applications .

P04626 REA mRNA status contributes to the discrepancy between gene amplification and protein overexpression in gastric cancer . BACKGROUND : Human epidermal growth factor receptor 2 ( P04626 REA ) is an important proto-oncogene of prognostic use in gastric cancer ( GC ) . Fluorescence in-situ hybridization ( Q5TCZ1 ) and immunohistochemistry ( IHC ) are the main clinical methods of detection of P04626 REA , but consistency between the methods is poor and the cause of the discrepancy is unclear . AIM : To investigate the involvement of P04626 REA mRNA status in the disparity between gene amplification and protein overexpression . METHODS : We investigated P04626 REA gene , mRNA , and protein profiles in gastric precancer and cancer tissues by use of the molecular approaches Q5TCZ1 , real-time polymerase chain reaction , and IHC . The relationships between P04626 REA and matrix metalloproteinase 9 ( P14780 REA ) and O15105 REA expression were analyzed and the involvement of P04626 REA in the interaction between tumor cells and lymphocytes was investigated by coculturing GC cell lines with peripheral blood mononuclear cells ( PBMCs ) . RESULTS : P04626 REA protein expression was significantly increased in cancer compared with precancer ( P = 0.003 ) , and the corresponding mRNA levels were significantly lower in precancer and cancer tissues than in normal tissues ( κ = 0.290 , P = 0.025 ) . P04626 REA mRNA levels were significantly higher in tumor than in peritumor tissue ( P = 0.028 ) , and were positively correlated with P14780 REA and O15105 REA mRNA levels in tumor tissues . P04626 REA mRNA expression in GC cell lines was increased by coculture with PBMCs . CONCLUSIONS : Different P04626 REA mRNA profiles , possibly in relation to contact between tumor cells and lymphocytes , might help to explain the discrepancy between gene amplification and protein overexpression results .

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 .

DB06273 MEN infusion therapy normalizes inflammation in sporadic P35858 REA patients . Patients with sporadic amyotrophic lateral sclerosis ( sALS ) show inflammation in the spinal cord and peripheral blood . The inflammation is driven by stimulation of macrophages by aggregated superoxide dismutase 1 ( P00441 REA ) through caspase 1 , interleukin 1 ( IL1 ) , P05231 REA and chemokine signaling . Inflammatory gene activation is inhibited in vitro by tocilizumab , a humanized antibody to P05231 REA receptor ( P08887 REA ) . DB06273 MENMAX DB06273 MEN inhibits global interleukin - 6 ( P05231 REA ) signaling , a key mechanism in chronic rheumatoid disorders . Here we studied in vivo baseline inflammatory gene transcription in peripheral blood mononuclear cells ( PBMCs ) of 10 sALS patients , and the effects of tocilizumab ( Actemra ( R ) ) infusions . At baseline , one half of P35858 REA subjects had strong inflammatory activation ( Group 1 ) ( 8 genes up regulated > 4 - fold , P < 0.05 vs . controls ) and the other half ( Group 2 ) had weak activation . All patients showed greater than four-fold up regulation of P03956 REA , P8 0098 , Q99616 REA and O00175 . DB06273 MEN infusions in the Group 1 patients resulted in down regulation of inflammatory genes ( in particular IL1β ) , whereas in the Group 2 patients in up regulation of inflammatory genes . Post-infusion serum and P04141 REA concentrations of tocilizumab inhibited caspase 1 activation in vitro . Three of 5 patients receiving tocilizumab infusions showed time-limited attenuation of clinical progression . In conclusion , inflammation of sALS patients at baseline is up - or down-regulated in comparison to controls , but is partially normalized by tocilizumab infusions .

Interrelationship between signal transduction pathways and 1,25 ( OH ) 2D3 in UMR 106 osteoblastic cells . In this study , the interrelationship between signal transduction pathways and 1,25- dihydroxyvitamin D ( 3 ) [ 1,25 ( OH ) 2D3 ] action was examined in UMR 106 osteoblastic cells . Treatment of these cells with 8 - bromo - DB02527 ( 1 mM ) resulted in an upregulation of the vitamin D receptor ( P11473 REA ) and an augmentation in the induction by 1,25 ( OH ) 2D3 of DB00146 MEN 24 - hydroxylase [ 24 ( OH ) ase ] and osteopontin ( P10451 REA ) mRNAs as well as gene transcription . Transfection with constructs containing the vitamin D response element devoid of other promoter regulatory elements did not alter the DB02527 - mediated potentiation , suggesting that DB02527 - enhanced transcription is due , at least in part , to upregulation of P11473 REA . Treatment with phorbol ester [ 12 - O-tetradecanoyl-phorbol - 13 - acetate ( TPA ) 100 nM ] , an activator of protein kinase C , significantly enhanced 1,25 ( OH ) 2D3 - induced P10451 REA mRNA and transcription but had no effect on P11473 REA or on 24 ( OH ) ase mRNA or transcription . Studies using P10451 REA promoter constructs indicate that TPA-enhanced P10451 REA transcription is mediated by an effect on the P10451 REA promoter separate from an effect on P11473 REA . Thus interactions with signal transduction pathways can enhance 1,25 ( OH ) 2D3 induction of 24 ( OH ) ase and P10451 REA gene expression , and , through different mechanisms , changes in cellular phosphorylation may play a significant role in determining the effectiveness of 1,25 ( OH ) 2D3 on transcriptional control in cells expressing skeletal phenotypic properties .

An osteopontin promoter polymorphism is associated with aggressiveness in breast cancer . Metastasis-related genes are deregulated in cancer by aberrant expression or splicing . Here , we analyzed polymorphic sites in the osteopontin promoter as potential contributors to aberrant expression in breast cancers . This study comprised 241 breast cancer specimens , for which DNA from normal surrounding tissue was available for 111 , and 65 healthy breast samples . The polymorphic site in position - 443 of the promoter was associated with tumor grade . As expected , there was no association between promoter single nucleotide polymorphisms ( SNPs ) and tumor stage or in situ carcinoma versus cancer , as stage and early transformation are determined by the sampling time more than by tumor genetics . In a subset of samples , osteopontin RNA expression levels had previously been obtained . The allelic distribution in positions - 443 and - 1748 was distinct between high and low expressors , confirming the importance of promoter SNPs . These two sites also form a haplotype . P10451 REA expression has been associated with breast cancer progression , regardless of the histological subtype of the cancer . Remarkably , the polymorphic site at - 443 , but not - 1748 or - 1776 , showed differences between ER-positive and ER-negative breast cancers and between PR-positive and PR-negative breast cancers , but there was no association with P04626 REA status . In five cases , the genotype of the tumor was different from the genotype of the host , implying the possibility of somatic mutations in the osteopontin promoter that may affect expression . Our results corroborate that the osteopontin promoter SNPs - 443 ( rs11730582 ) and - 1748 ( rs2728127 ) are important for gene expression and breast cancer aggressiveness .

[ DB00594 MEN attenuates hypoxia-induced proliferation of rats pulmonary artery smooth muscle cells by suppressing Na + / H + exchanger - 1 ] . AIM : To study the influence of Na + / H + exchange inhibitor amiloride on hypoxia-induced proliferation in rats pulmonary artery smooth muscle cells ( PASMCs ) , also observe the change of Na + / H + exchanger - 1 ( P19634 REA ) activity and expression . METHODS : Rats PASMGs were cultured in normoxia ( 21 % O2 ) or hypoxia ( 2 % O2 ) for 24 hours , as well as administered amiloride with various concentrations , cultured for 24 hours , then determined MTT OD values and rates of P12004 REA positive cells to investigate cells proliferation , moreover intracellular pH was determined by interactive Laser Cytometer , and Na + / H + exchanger - 1 mRNA expression was determined by RT-PCR . RESULTS : Hypoxic exposure heightened intracellular pH and mRNA expression of P19634 REA in PASMCs , however , 3.123- 50 micromol / L amiloride depressed them gradually . Additionally , hypoxic exposure raised MTT OD value and rates of P12004 REA positive cells , similarly , the above two indexes descended gradually with presence of 3.125- 50 micromol / L amiloride . CONCLUSION : Na + / H + exchange inhibitor amiloride can suppress hypoxia-induced proliferation in pulmonary artery smooth muscle cells , which is due to depress activity and expression of P19634 REA .

Lack of biological relevance of platelet cyclooxygenase - 2 dependent thromboxane A2 production . INTRODUCTION : There is emerging evidence of a considerable variability of the impact of aspirin on clinical outcome and laboratory findings . Persistent TxA 2 production seems to be the most likely reason . Aim of this study was to determine whether the mechanism responsible for TxA 2 persistent production is , at least partially , dependent upon aspirin-insensitive platelet P35354 REA enzymatic pathway . METHODS AND RESULTS : In 100 consecutive patients , under chronic aspirin anti-platelet treatment ( 100-160 mg / day ) selected on the basis of detectable plasma salicylate levels , serum and DB04557 MEN ( AA ) - induced platelet TxA 2 production , immunoblot analysis of platelet P23219 REA / P35354 REA expression and P35354 REA activity were studied . Immunoblot revealed P35354 REA expression in 46 % patients , in an amount that was markedly lower than P23219 REA . In 10 P35354 REA positive patients with TxA 2 levels over the median , AA-induced TxA 2 production performed in vitro in the presence of the P35354 REA inhibitor CAY 10404 and aspirin demonstrated that P35354 REA dependent TxA 2 production is less than 2 % . CONCLUSION : Our data demonstrate that the inter-individual variability of platelet sensitivity to aspirin is due to a reduced efficacy of aspirin on platelet P23219 REA despite ascertained patient compliance . We suggest that serum TxA 2 assay might be performed in future clinical studies to improve our knowledge on the residual TxA 2 production in aspirin-treated patients .

P61968 is an essential mediator of ErbB 2 / P04626 REA / Neu-induced breast cancer cell cycle progression . ErbB 2 / P04626 REA / Neu-overexpressing breast cancers are characterized by poor survival due to high proliferation and metastasis rates and identifying downstream targets of ErbB 2 should facilitate developing novel therapies for this disease . Gene expression profiling revealed the transcriptional regulator LIM-only protein 4 ( P61968 ) is upregulated during ErbB 2 - induced mouse mammary gland tumorigenesis . Although P61968 is frequently overexpressed in breast cancer and P61968 - overexpressing mice develop mammary epithelial tumors , the mechanisms involved are unknown . In this study , we report that P61968 is a downstream target of ErbB 2 and PI3K in ErbB 2 - dependent breast cancer cells . Furthermore , P61968 silencing reduces proliferation of these cells , inducing a G2 / M arrest that was associated with decreased cullin - 3 , an E3 - ubiquitin ligase component important for mitosis . Loss of P61968 subsequently results in reduced P12004 REA D1 and P12004 REA E . Further supporting a role for P61968 in modulating proliferation by regulating cullin - 3 expression , we found that P61968 expression oscillates throughout the cell cycle with maximum expression occurring during G2 / M and these changes precede oscillations in cullin - 3 levels . P61968 levels are also highest in high-grade / less differentiated breast cancers , which are characteristically highly proliferative . We conclude that P61968 is a novel cell cycle regulator with a key role in mediating ErbB 2 - induced proliferation , a hallmark of ErbB 2 - positive disease .

Hypoxic / normoxic preconditioning increases endothelial differentiation potential of human bone marrow CD133 + cells . CD133 + cells are hemangioblasts that have capacity to generate into both hematopoietic and endothelial cells ( ECs ) . Hypoxia / normoxia has shown to be the regulator of the balance between stemness and differentiation . In this study we performed Agilent ' s whole human genome oligo microarray analysis and examined the differentiation potential of the bone-marrow-derived CD133 + cells after hypoxic / normoxic preconditioning of CD133 + cells . Results showed that there was no significant increase in erythroid colony forming unit ( CFU-E ) and CFU-granulocyte , erythrocyte , monocyte , and megakaryocyte formation with cells treated under hypoxia / normoxia . However , a significant increment of EC forming unit at 24 h ( 143.2 + / - 8.0 % ) compared to 0 h ( 100 + / - 11.4 % ) was observed in CFU-EC analysis . Reverse transcription-polymerase chain reaction and immunostaining analysis showed that the differentiated cells diminished hematopoietic stem cell surface markers and acquired the gene markers and functional phenotype of ECs . The transcriptome profile revealed a cluster of 232 downregulated and 498 upregulated genes in cells treated for 24 h under hypoxia . The upregulated genes include angiogenic genes , angiogenic growth factor genes , angiogenic cytokine and chemokine genes , as well as angiogenic-positive regulatory genes , including Q14512 REA , PDGFB , Q16663 , P48061 REA , P8 0162 , P05231 REA , P21246 , O14944 REA , P04626 REA , O95136 REA , P11487 REA , Q92913 REA , Q99988 , P05412 REA , L1CAM , Q02297 REA , P08138 REA , and PDGFB . On the other hand , angiogenesis inhibitors and related genes , including P29459 REA , P98177 REA , Q9NY15 , and P16035 REA , are downregulated . Taken together , hypoxic / normoxic preconditioning may lead to the differentiation of CD133 + cells toward endothelial lineage , which may improve the current clinical trial studies .