interacts_with DB09280?
Repeated cisplatin treatment can lead to a multiresistant tumor cell population with stem cell features and sensitivity to 3-bromopyruvate . DB00515 is used in treatment of several types of cancer , including epithelial ovarian carcinoma ( EOC ) . In order to mimic clinical treatment and to investigate longterm effects of cisplatin in surviving cancer cells , two EOC cell lines were repeatedly treated with low doses . In the SKOV-3 cell line originating from malignant ascites , but not in A2780 cells from a primary tumor , this led to emergence of a stable population ( SKOV-3-R ) which in the absence of cisplatin showed increased motility , epithelial-mesenchymal transition ( EMT ) and expression of cancer stem cell markers CD117 , P16070 and P00352 . Accordingly , the cells formed self-renewing spheres in serum-free stem cell medium . Despite upregulation of mitochondrial mass and cytochrome c , and no upregulation of Bcl-2/Bcl-xL , SKOV-3-R were multiresistant to antineoplastic drugs . Cancer stem cells , or tumor-initiating cells ( TICs ) are highly chemoresistant and are believed to cause relapse into disseminated and resistant EOC . Our second aim was therefore to target resistance in these Q8NDX1 -like cells . Resistance could be correlated with upregulation of hexokinase-II and P21796 , which are known to form a survival-promoting mitochondrial complex . The cells were thus sensitive to 3-bromopyruvate , which dissociates hexokinase-II from this complex , and were particularly sensitive to combination treatment with cisplatin at doses down to 0.1 x IC 50 . 3-bromopyruvate might thus be of use in targeting the especially aggressive Q8NDX1 populations . Potentiator ivacaftor abrogates pharmacological correction of ΔF508 P13569 in cystic fibrosis . Cystic fibrosis ( CF ) is caused by mutations in the CF transmembrane conductance regulator ( P13569 ) . Newly developed " correctors " such as DB09280 ( VX-809 ) that improve P13569 maturation and trafficking and " potentiators " such as ivacaftor ( VX-770 ) that enhance channel activity may provide important advances in CF therapy . Although VX-770 has demonstrated substantial clinical efficacy in the small subset of patients with a mutation ( G551D ) that affects only channel activity , a single compound is not sufficient to treat patients with the more common P13569 mutation , ΔF508 . Thus , patients with ΔF508 will likely require treatment with both correctors and potentiators to achieve clinical benefit . However , whereas the effectiveness of acute treatment with this drug combination has been demonstrated in vitro , the impact of chronic therapy has not been established . In studies of human primary airway epithelial cells , we found that both acute and chronic treatment with VX-770 improved P13569 function in cells with the G551D mutation , consistent with clinical studies . In contrast , chronic VX-770 administration caused a dose-dependent reversal of VX-809-mediated P13569 correction in ΔF508 homozygous cultures . This result reflected the destabilization of corrected ΔF508 P13569 by VX-770 , markedly increasing its turnover rate . Chronic VX-770 treatment also reduced mature wild-type P13569 levels and function . These findings demonstrate that chronic treatment with P13569 potentiators and correctors may have unexpected effects that can not be predicted from short-term studies . Combining these drugs to maximize rescue of ΔF508 P13569 may require changes in dosing and/or development of new potentiator compounds that do not interfere with P13569 stability . Endogenous concentrations of ouabain act as a cofactor to stimulate fluid secretion and cyst growth of in vitro ADPKD models via DB02527 and P00533 -Src-MEK pathways . In autosomal-dominant polycystic kidney disease ( ADPKD ) , renal cysts develop by aberrant epithelial cell proliferation and transepithelial fluid secretion . We previously showed that ouabain increases proliferation of cultured human ADPKD cells via stimulation of the P01133 receptor ( P00533 ) -Src-MEK/ P29323 signaling pathway . We examined whether ouabain affects fluid secretion and in vitro cyst growth of human ADPKD cell monolayers , ADPKD cell microcysts cultured in a three-dimensional collagen matrix , and metanephric organ cultures from Pkd1(m1Bei) mice . Physiological concentrations of ouabain alone did not affect net transepithelial basal-to-apical fluid transport in ADPKD monolayers or growth of cultured ADPKD microcysts . In contrast , in the presence of forskolin or 8-bromo- DB02527 , ouabain significantly enhanced ADPKD fluid secretion and microcyst expansion . Ouabain exerted this effect by enhancing DB02527 -dependent Cl(-) secretion via the P13569 . Similarly , ouabain accelerated DB02527 -dependent cyst enlargement in Pkd1(m1Bei) mice metanephroi , with a more prominent response in homozygous than heterozygous mice . Ouabain had no effect on fluid secretion and cystogenesis of normal human kidney cells and caused only slight cystic dilations in wild-type mouse kidneys . The effects of ouabain in ADPKD cells and Pkd1(m1Bei) metanephroi were prevented by inhibitors of P00533 ( AG1478 ) , Src ( Q99463 ) , and MEK ( U0126 ) . Together , our results show that ouabain , used in physiological concentrations , has synergistic effects on DB02527 -mediated fluid secretion and cyst growth , via activation of the P00533 -Src-MEK pathway . These data provide important evidence for the role of ouabain as an endogenous hormone that exacerbates ADPKD cyst progression . Expression of the low-density lipoprotein receptor , P04035 , and multidrug resistance ( Mdr1 ) genes in colorectal carcinomas . Some malignant cells have elevated uptake of plasma low-density lipoprotein ( LDL ) . We determined the expressions in colorectal cancers of the P01130 gene , of the gene for the rate-limiting enzyme in cholesterol synthesis , 3-hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase , and of the multidrug resistance gene ( mdr1 ) by quantitative RNA-RNA solution hybridisation . P01130 RNA levels in tumor tissue exceeded those in normal mucosa in 20 of 23 patients ( 2-11-fold higher in 17 of 23 patients ) , with a mean +/- SD of 7.8 +/- 5.8 copies/cell in tumor tissue vs 3.5 +/- 2.5 in normal mucosa ( P = 0.002 ) . The P04035 gene was similarly expressed in tumor and normal tissue , with means and SD of 2.0 +/- 1.3 copies/cell versus 2.2 +/- 1.9 ( pi = 21 ) . Mdr1 RNA was undetectable ( < 0.15 copies/cell ) in 5 of 20 tumors , with a mean +/- SD of 1.0 +/- 1.1 copies/cell vs 1.6 +/- 1.7 in normal mucosa . Expression of all three genes was , in most cases , higher in normal liver than in liver metastasis of colorectal carcinomas or normal colon mucosa . The results may form the basis for using LDL as a drug carrier for treatment of colorectal carcinomas , and may indicate that drug resistance in these tumors is not due to overexpression of the mdr1 gene . Selective use of multiple vitamin D response elements underlies the 1 alpha,25-dihydroxyvitamin D3-mediated negative regulation of the human O15528 gene . The human 25-hydroxyvitamin D3 ( DB00146 ) 1alpha-hydroxylase , which is encoded by the O15528 gene , catalyzes the metabolic activation of the DB00146 into 1alpha,25-dihydroxyvitamin D3 ( DB00136 ) , the most biologically potent vitamin D3 metabolite . The most important regulator of O15528 gene activity is DB00136 itself , which down-regulates the gene . The down-regulation of the O15528 gene has been proposed to involve a negative vitamin D response element ( nVDRE ) that is located approximately 500 bp upstream from transcription start site ( TSS ) . In this study , we reveal the existence of two new P11473 -binding regions in the distal promoter , 2.6 and 3.2 kb upstream from the TSS , that bind vitamin D receptor-retinoid X receptor complexes . Since the down regulation of the O15528 gene is tissue- and cell-type selective , a comparative study was done for the new DB00136 -responsive regions in P29320 -293 human embryonic kidney and MCF-7 human breast cancer cells that reflect tissues that , respectively , are permissive and non-permissive to the phenomenon of DB00136 -mediated down-regulation of this gene . We found significant differences in the composition of protein complexes associated with these O15528 promoter regions in the different cell lines , some of which reflect the capability of transcriptional repression of the O15528 gene in these different cells . In addition , chromatin architecture differed with respect to chromatin looping in the two cell lines , as the new distal regions were differentially connected with the proximal promoter . This data explains , in part , why the human O15528 gene is repressed in P29320 -293 but not in MCF-7 cells . Multiple endocytic signals in the C-terminal tail of the cystic fibrosis transmembrane conductance regulator . The cystic fibrosis transmembrane conductance regulator ( P13569 ) is a DB02527 -dependent protein kinase ( PKA ) -activated chloride channel that is localized to the plasma membrane and endosomal compartment . Endosomal targeting of P13569 is attributed to the DB00135 (1424)-based internalization signal , identified in the C-terminal tail of the channel . Mutation of the DB00135 (1424) residue could partly inhibit the endocytosis of P13569 and its association with the adapter protein P05549 . To reveal additional endosomal targeting signals , site-directed mutagenesis of both a chimaera , composed of a truncated form of interleukin 2 receptor alpha chain ( TacT ) and the C-terminal tail of P13569 ( Ct ) , and the full-length P13569 was performed . Morphological and functional assays revealed the presence of multiple internalization motifs at the C-terminus , consisting of a phenylalanine-based motif ( DB00120 (1413) ) and a bipartite endocytic signal , comprising a tyrosine ( DB00135 (1424) ) and a di- DB00149 -based ( DB00149 (1430)- DB00149 ) motif . Whereas the replacement of any one of the three internalization motifs with alanine prevented the endocytosis of the TacT-Ct chimaera , mutagenesis of DB00120 (1413)- DB00149 impaired the biosynthetic processing of P13569 , indicating that DB00120 (1413) is indispensable for the native structure of P13569 . In contrast , replacement of DB00149 (1430)- DB00149 - and DB00135 (1424)-based signals with alanine increased the cell-surface density of both the chimaeras and P13569 in an additive manner . These results suggest that the internalization of P13569 is regulated by multiple endocytic sorting signals . Preclinical rationale for combining an P00533 antibody with cisplatin/gemcitabine for the treatment of NSCLC . BACKGROUND : Although the addition of epidermal growth factor receptor ( P00533 ) antibodies to various platinum-based chemotherapy regimens for non-small cell lung cancer ( NSCLC ) is being actively pursued in the clinic , rationale for the prioritization of specific regimens is lacking . MATERIALS AND METHODS : We evaluated the antitumor effects of necitumumab , a recombinant human IgG1 antibody targeting P00533 , in combination with cisplatin plus gemcitabine , pemetrexed , or paclitaxel in a panel of 9 subcutaneous tumor models of NSCLC established in nu/nu athymic mice . RESULTS : DB09559 in combination with cisplatin/gemcitabine was particularly effective , although interestingly , the mechanisms underlying these benefits were model dependent . For example , increased tumor cell apoptosis contributed towards combination efficacy in the A549 model , in association with increased expression of hsa-miR-29b and reduced expression of antiapoptotic genes including DNA methyltransferase Q9UBC3 , commonly up-regulated in patients with NSCLC . Such inverse effects of combination therapy on Q9UBC3 and hsa-miR-29b expression were found in multiple models . Importantly , in the A549 model , hsa-miR-29b down-regulation of DMNT3b reduced promoter methylation of tumor suppressor genes such as Q9BY67 ( Q9BY67 ) , Ras associated ( Q12967 /AF-6 ) domain family member 1 ( Q9NS23 ) , and Fragile histidine triad gene ( P49789 ) , increasing their expression . CONCLUSION : These results offer a preclinical rationale for combining an P00533 antibody with cisplatin/gemcitabine for patients with NSCLC , and provide potential molecular biomarkers for tailoring therapy . Regulation of male fertility by P13569 and implications in male infertility . BACKGROUND : The cystic fibrosis transmembrane conductance regulator ( P13569 ) is a DB02527 -activated Cl(-) and HCO(3)(-) conducting channel , mutations of which are known to be associated with male infertility . However , the underlying mechanisms remain elusive . METHODS : Literature databases were searched for papers on the topics related to P13569 and male fertility and infertility with relevant keywords . Unpublished data from authors ' laboratory were also included for analysis . RESULTS : Clinical evidence shows increased mutation frequency or reduced P13569 expression in men with congenital bilateral absence of vas deferens ( CBAVD ) or sperm abnormalities , such as azoospermia teratospermia and oligoasthenospermia . Studies on primary rodent Sertoli cells and germ cells , as well as testes from P13569 knockout mice or a cryptorchidism model , yield findings indicating the involvement of P13569 in spermatogensis through the HCO(3)(-)/ Q96PN6 / DB02527 /CREB( Q03060 ) pathway and the NF-κB/ P35354 /PGE(2) pathway . Evidence also reveals a critical role of P13569 in sperm capacitation by directly or indirectly mediating HCO(3)(-) entry that is essential for capacitation . P13569 is emerging as a versatile player with roles in mediating different signaling pathways pertinent to various reproductive processes , in addition to its long-recognized role in electrolyte and fluid transport that regulates the luminal microenvironment of the male reproductive tract . CONCLUSIONS : P13569 is a key regulator of male fertility , a defect of which may result in different forms of male infertility other than CBAVD . It would be worthwhile to further investigate the potential of developing novel diagnostic and contraceptive methods targeting P13569 . Telomere shortening is associated with reduced duodenal HCOFormula secretory but normal gastric acid secretory capacity in aging mice . The incidence of duodenal ulcer , especially Helicobacter pylori-negative duodenal ulcer , strongly increases with age . In humans , telomere length shortening is considered to be one critical factor in cellular senescence and organ survival . In this study , we compared basal and stimulated gastric acid and duodenal HCO(3)(-) secretory rates in aged late-generation ( G(3) ) telomerase-deficient ( mTERC(-/-) ) mice , which are characterized by severe telomere dysfunction due to the inability to elongate telomeres during cell division . We found that basal and forskolin-stimulated HCO(3)(-) secretion and short-circuit current ( I(sc) ) in isolated duodenal mucosa of G(3) mTERC(-/-) mice were markedly reduced compared with age-matched wild-type mice . In contrast , basal and forskolin-stimulated acid secretory rates in isolated G(3) mTERC(-/-) gastric mucosa were not significantly altered . Correspondingly , duodenal mucosa of G(3) mTERC(-/-) mice showed slimming and shortening of villi , whereas gastric mucosal histology was not significantly altered . However , the ratios of cystic fibrosis transmembrane conductance regulator ( P13569 ) and solute-linked carrier 26 gene family ( Slc26a6 ) mRNA expression in relation to cytokeratin-18 were not altered in duodenal mucosa . The further knockout of P38936 , which is a downstream effector of telomere shortening-induced senescence , rescued villus atrophy of duodenal mucosa , and basal and forskolin-stimulated duodenal HCO(3)(-) secretion and I(sc) in mTERC(-/-) P38936 (-/-) double-knockout mice were not different from wild-type controls . In conclusion , genetic ablation of telomerase resulted in P38936 -dependent duodenal mucosal atrophy and reduced duodenal HCO(3)(-) secretory capacity , whereas gastric morphology and acid secretory function were preserved . This suggests that telomere shortening during aging may result in an imbalance between aggressive and protective secretions against duodenal mucosa and thus predispose to ulcer formation . Interplay between inhibitory ferric and stimulatory curcumin regulates phosphorylation-dependent human cystic fibrosis transmembrane conductance regulator and ΔF508 activity . Curcumin potentiates cystic fibrosis transmembrane conductance regulator ( P13569 ) activation in an DB00171 -independent but phosphorylation-dependent manner . The underlying molecular mechanisms are unclear . Here , P29320 -293T cells cultured in an Fe(3+)-containing medium were transiently transfected with P13569 constructs , and the role of the inhibitory Fe(3+) bridge between intracellular loop 3 and the regulatory domain of P13569 in this pathway was investigated . The results showed that ethylenediaminetetraacetic acid ( DB00974 ) stimulated phosphorylation-dependent P13569 activation and the stimulation was suppressed by the deletion of the regulatory domain or the insertion of a C832A mutation that removes the Fe(3+)-binding interface . Furthermore , curcumin potentiation of P13569 was significantly weakened not only by Fe(3+)-insensitive mutations at the interface between the regulatory domain and intracellular loop 3 but also by N-ethylmaleimide or DB00974 pretreatment that removes Fe(3+) . More importantly , potentiation of P13569 was completely suppressed by sufficient Fe(3+) . Finally , the insertion of Fe(3+)-insensitive H950R/S768R increased the curcumin-independent activity of ΔF508 but weakened its curcumin potentiation . Thus , Fe(3+) homeostasis in epithelia may play a critical role in regulating P13569 activity , and targeting Fe(3+)-chelating potentiators may direct new therapies for cystic fibrosis . Caenorhabditis elegans expressed sequence tags identify gene families and potential disease gene homologues . A database containing mapped partial cDNA sequences from Caenorhabditis elegans will provide a ready starting point for identifying nematode homologues of important human genes and determining their functions in C. elegans . A total of 720 expressed sequence tags ( ESTs ) have been generated from 585 clones randomly selected from a mixed-stage C. elegans cDNA library . Comparison of these ESTs with sequence databases identified 422 new C. elegans genes , of which 317 are not similar to any sequences in the database . Twenty-six new genes have been mapped by YAC clone hybridization . Members of several gene families , including cuticle collagens , GTP-binding proteins , and RNA helicases were discovered . Many of the new genes are similar to known or potential human disease genes , including P13569 and the P01130 . Osteopetrosis : genetics , treatment and new insights into osteoclast function . Osteopetrosis is a genetic condition of increased bone mass , which is caused by defects in osteoclast formation and function . Both autosomal recessive and autosomal dominant forms exist , but this Review focuses on autosomal recessive osteopetrosis ( ARO ) , also known as malignant infantile osteopetrosis . The genetic basis of this disease is now largely uncovered : mutations in Q13488 , P51798 , Q86WC4 , Q9Y5X0 and Q9Y4G2 lead to osteoclast-rich ARO ( in which osteoclasts are abundant but have severely impaired resorptive function ) , whereas mutations in O14788 and Q9Y6Q6 lead to osteoclast-poor ARO . In osteoclast-rich ARO , impaired endosomal and lysosomal vesicle trafficking results in defective osteoclast ruffled-border formation and , hence , the inability to resorb bone and mineralized cartilage . ARO presents soon after birth and can be fatal if left untreated . However , the disease is heterogeneous in clinical presentation and often misdiagnosed . This article describes the genetics of ARO and discusses the diagnostic role of next-generation sequencing methods . The management of affected patients , including guidelines for the indication of haematopoietic stem cell transplantation ( which can provide a cure for many types of ARO ) , are outlined . Finally , novel treatments , including preclinical data on in utero stem cell treatment , O14788 replacement therapy and denosumab therapy for hypercalcaemia are also discussed . Vitamin D metabolism , mechanism of action , and clinical applications . DB00169 is made in the skin from 7-dehydrocholesterol under the influence of UV light . DB00153 ( ergocalciferol ) is derived from the plant sterol ergosterol . Vitamin D is metabolized first to 25 hydroxyvitamin D ( 25OHD ) , then to the hormonal form 1,25-dihydroxyvitamin D ( 1,25(OH)2D ) . Q6VVX0 is the most important 25-hydroxylase ; O15528 is the key 1-hydroxylase . Both 25OHD and 1,25(OH)2D are catabolized by Q07973 . 1,25(OH)2D is the ligand for the vitamin D receptor ( P11473 ) , a transcription factor , binding to sites in the DNA called vitamin D response elements ( VDREs ) . There are thousands of these binding sites regulating hundreds of genes in a cell-specific fashion . P11473 -regulated transcription is dependent on comodulators , the profile of which is also cell specific . Analogs of 1,25(OH)2D are being developed to target specific diseases with minimal side effects . This review will examine these different aspects of vitamin D metabolism , mechanism of action , and clinical application . Differential regulation of chondrogenic differentiation by the serotonin2B receptor and retinoic acid in the embryonic mouse hindlimb . Retinoic acid ( RA ) synthesizing and metabolizing enzymes are coordinately expressed with serotonin 2B ( P41595 ) receptors at sites of epithelial-mesenchymal ( E-M ) interaction in the mouse embryo ( Bhasin et al. , 1999 ) . The promoter of the P41595 receptor contains potential RA response element ( RAREs ) as well as an P05549 site . Because both retinoid and serotonergic signaling have been implicated in the regulation of chondrogenic differentiation , the present study investigated whether these signals may work together to regulate this morphogenetic process in hindlimb bud micromass cultures . Results indicate that 5-HT promotes [35S]sulfate incorporation ( chondrogenic differentiation ) by activation of P41595 receptors , which use the mitogen activated protein kinase ( Q8NFH3 MAPK ) signal transduction pathway , whereas RA dose-dependently inhibits sulfate incorporation and promotes expression of RARbeta , which could lead to inhibition of p38 MAPK . No evidence was found to support the possibility that RA negatively regulates expression of P41595 receptors . Taken together , these results suggest that 5-HT and RA may act as opposing signals to regulate chondrogenic differentiation in the developing hindlimb , possibly mediated by different MAPK signal transduction pathways . Protective effect of treatment with low-dose gliclazide in a model of middle cerebral artery occlusion and reperfusion in rats . The aim of this study was to explore the expression of sulfonylurea receptor 1 ( Q09428 ) , the regulatory subunit of the NCCa- DB00171 channel , and to investigate the protective effects of gliclazide following middle cerebral artery occlusion ( MCAO ) /reperfusion in male Wistar rats . Adult rats underwent 2h of the left MCAO using the intraluminal thread technique before reperfusion . The core areas of the infarct at different reperfusion time points were examined for the mRNA level and protein expression of Q09428 using reverse transcription-polymerase chain reaction ( RT-PCR ) and western blotting respectively . DB01120 was administered intravenously into the right jugular vein for 12h simultaneously with the reperfusion . The number of apoptotic cells was determined using the TUNEL assay . The neurological functional deficits were evaluated using Bederson׳s test , and the cerebral infarction volume was visualized with TTC staining . We found up-regulation of Q09428 mRNA and protein levels in ischemic infarct tissues after reperfusion following MCAO , and Q09428 mRNA and protein were maximally upregulated 8-12h after a 2-hour ischemia . The treatment with low-dose of gliclazide reduced the total number of TUNEL-positive cells , the neurological functional deficits and the brain infarct volume . These results suggest that the Q09428 -regulated NCCa- DB00171 channel may be associated with MCAO/reperfusion injury and the infarct-reducing effects of intravenous treatment with gliclazide may be due , in part , to the blocked upregulation of Q09428 expression , the decreased infarct size and the reduced apoptosis in the ischemia-reperfusion brain . Acute ethanol preexposure promotes liver regeneration after partial hepatectomy in mice by activating P05091 . It is known that chronic ethanol significantly impairs liver regeneration . However , the effect of acute ethanol exposure on liver regeneration remains largely unknown . To address this question , C57Bl6/J mice were exposed to acute ethanol ( 6 g/kg intragastrically ) for 3 days , and partial hepatectomy ( PHx ) was performed 24 h after the last dose . Surprisingly , acute ethanol preexposure promoted liver regeneration . This effect of ethanol did not correlate with changes in expression of cell cycle regulatory genes ( e.g. , cyclin D1 , P38936 , and p27 ) but did correlate with protection against the effect of PHx on indices of impaired lipid and carbohydrate metabolism . DB00898 preexposure protected against inhibition of the oxidant-sensitive mitochondrial enzyme , aconitase . The activity of aldehyde dehydrogenase 2 ( P05091 ) was significantly increased by ethanol preexposure . The effect of ethanol was blocked by inhibiting ( DB02115 ) and was mimicked by activating ( Alda-1 ) P05091 . Lipid peroxides are also substrates for P05091 ; indeed , alcohol preexposure blunted the increase in lipid peroxidation ( 4OH-nonenal adducts ) caused by PHx . Taken together , these data suggest that acute preoperative ethanol exposure " preconditions " the liver to respond more rapidly to regenerate after PHx by activating mitochondrial P05091 , which prevents oxidative stress in this compartment . Biophysical and pharmacological characterization of hypotonically activated chloride currents in cortical astrocytes . Rat cortical astrocytes regulate their cell volume in response to hypotonic challenge . This regulation is believed to depend largely on the release of chloride or organic osmolytes through anion channels . Using whole-cell recordings , we identified weakly outwardly rectifying chloride currents that could be activated in response to hypotonic challenge . These currents exhibited the following permeability sequence upon replacement of chloride in the bathing solution with various anions : I- > NO3- > Cl- > Gluc- > or = MeS- > Ise- . Interestingly , extracellular I- , albeit showing the greatest permeability , blocked the currents with an IC50 of approximately 50 mM . Currents were almost completely inhibited by 123 microM P16860 and partially inhibited by 200 microM niflumic acid or 200 microM DIDS . Additionally , the total number of Cl- ions effluxed through the hypotonically activated channels was markedly similar to the total solute efflux during volume regulation . We therefore propose the hypotonically activated chloride channel as a major contributor to volume regulation of astrocytes . To examine potential candidate chloride channel genes expressed by astrocytes , we employed RT-PCR to demonstrate the presence of transcripts for P51788 , 3 , 4 , 5 , and 7 , as well as for P21796 and P13569 in cultured astrocytes . Moreover , we performed immunostaining with antibodies against each of these channels and showed the strongest expression of P51788 and P51790 , strong expression of P51795 and P21796 , weak expression of P51798 and very weak expression of P51793 and P13569 . Intriguingly , although we found at least seven Cl- channel proteins from three different gene families in astrocytes , none appeared to be active in resting cells . DB05822 , a nitric oxide-releasing aspirin derivative , exhibits a significant antiproliferative effect and alters cell cycle progression in human colon adenocarcinoma cell lines . DB00435 -releasing non-steroidal antiinflammatory drugs ( NO-NSAIDs ) are safer than NSAIDs due to their ability to reduce gastric toxicity . We assessed the cytotoxic activity of a new aspirin derivative , DB05822 , after different exposure schedules , in three human colon adenocarcinoma cell lines . All the lines were positive for P23219 protein and mRNA , as evaluated by Western blot and RT-PCR , respectively , while only one was positive for P35354 . The cytostatic and cytocidal activity was determined by sulforhodamine B assay and evaluated according to Monks ' model . Cytostatic activity was observed after a 24-h drug exposure and 50 % growth inhibition was reached at concentrations ranging from 165 to 250 micro M in all cell lines , whereas with aspirin the IC50 was never reached , even at the maximum concentration tested ( 500 micro M ) , and was independent of P23219 or P35354 status . Cytocidal activity was observed only at the highest concentrations and persisted for a long time after drug removal . Flow cytometric analysis showed that the NO-aspirin compound induced a persistent accumulation of cells in G2-M phase in all the cell lines after at least 48 h exposure . Specifically , the block pertained mainly to G2 phase , whereas mitotic index was not affected at all . Our results indicate that DB05822 has an in vitro cytostatic activity superior to that of its parental aspirin compound , which makes it a potentially important tumor preventive agent . Furthermore , the cytocidal effect observed at the highest concentrations and the induction of a specific block in G2 phase renders it a promising candidate for drug combination treatments . DB11582 suppresses osteoclastogenesis induced by O14788 and cancer cells through inhibition of inflammatory pathways : a new use for an old drug . BACKGROUND AND PURPOSE : Most patients with cancer die not because of the tumour in the primary site , but because it has spread to other sites . Common tumours , such as breast , multiple myeloma , and prostate tumours , frequently metastasize to the bone . To search for an inhibitor of cancer-induced bone loss , we investigated the effect of thiocolchicoside , a semi-synthetic colchicoside derived from the plant Gloriosa superba and clinically used as a muscle relaxant , on osteoclastogenesis induced by receptor activator of NF-κB ligand ( O14788 ) and tumour cells . EXPERIMENTAL APPROACH : We used RAW 264.7 ( murine macrophage ) cells , a well-established system for osteoclastogenesis , and evaluated the effect of thiocolchicoside on O14788 -induced NF-κB signalling and osteoclastogenesis as well as on osteoclastogenesis induced by tumour cells . KEY RESULTS : DB11582 suppressed osteoclastogenesis induced by O14788 , and by breast cancer and multiple myeloma cells . Inhibition of the NF-κB pathway was responsible for this effect since the colchicoside inhibited O14788 -induced NF-κB activation , activation of IκB kinase ( IKK ) and suppressed inhibitor of NF-κBα ( IκBα ) phosphorylation and degradation , an inhibitor of NF-κB . Furthermore , an inhibitor of the IκBα kinase γ or NF-κB essential modulator , the regulatory component of the IKK complex , demonstrated that the NF-κB signalling pathway is mandatory for osteoclastogenesis induced by O14788 . CONCLUSIONS AND IMPLICATIONS : Together , these data suggest that thiocolchicoside significantly suppressed osteoclastogenesis induced by O14788 and tumour cells via the NF-κB signalling pathway . Thus , thiocolchicoside , a drug that has been used for almost half a century to treat muscle pain , may also be considered as a new treatment for bone loss . Differences in transcript levels of ABC transporters between pancreatic adenocarcinoma and nonneoplastic tissues . OBJECTIVES : The aim of this study was to evaluate transcript levels of all 49 human DB00171 -binding cassette transporters ( ABCs ) in one of the most drug-resistant cancers , namely , the pancreatic ductal adenocarcinoma ( PDAC ) . Association of ABCs levels with clinical-pathologic characteristics and P01116 mutation status was followed as well . METHODS : Tumors and adjacent nonneoplastic tissues were obtained from 32 histologically verified PDAC patients . The transcript profile of ABCs was assessed using quantitative real-time polymerase chain reaction with a relative standard curve . P01116 mutations in exon 2 were assessed by high-resolution melting analysis and sequencing . RESULTS : Most ABCs were deregulated in PDAC and 10 ABCs were associated with clinical-pathologic characteristics . P01116 mutations did not change the global expression profile of ABCs . CONCLUSIONS : The expression of ABC transporters was significantly deregulated in PDAC tumors when compared to nonmalignant tissues . The observed up-regulation of P21439 , O95342 , P33527 , O15438 , O15440 , Q5T3U5 , and Q9UNQ0 in tumors may contribute to the generally poor treatment response of PDAC . The up-regulation of O95477 , Q8IZY2 , and P45844 implicates a serious impairment of cellular cholesterol homeostasis in PDAC . On the other hand , the observed down-regulation of Q99758 , O95255 , P13569 , and Q09428 suggests a possible role of stem cells in the development and progression of PDAC . [ Larval stages of Ascaris lumbricoides : hyaluronan-binding capacity ] . DB08818 has important functions in inflammatory and tissue reparation processes . Owing to the varied strategies of the parasites to evade the host 's immune response , as well as the multiple functions and physiological importance of hyaluronic acid , the aim was to study the hyaluronan binding capacity by Ascaris lumbricoides larval stages . Larval concentrates were prepared by hatching A. lumbricoides eggs . The larvae were collected by the Baermann method . The test of serum soluble P16070 detection by Agregation Inhibition was modified . All the larval concentrates presented hyaluronan binding capacity . The obtained results allow to suppose the existence of an hyaluronic acid specific receptor in A. lumbricoides . This receptor eventually might compete with the usual receptors of the host . The parasite might use this mechanism to evade the immune response . Agonism at P41595 receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5-hydroxytryptamine2B ( P41595 ) receptors . To evaluate whether agonism at P41595 receptors is a phenomenon of the class of the ergolines , we studied P41595 receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC50 8.42 and 8.72 ) . DB01200 acted as a partial agonist ( pEC50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5-HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 receptors seems not to be a class effect of the ergolines .