MH_dev_147

Q9Y6X2 REA negatively regulates O14788 REA - mediated osteoclastogenesis directly in osteoclast precursors and indirectly via osteoblasts . Cytokine signaling via various transcription factors regulates receptor activator of nuclear factor ( NF ) - kappaB ligand ( O14788 REA ) - mediated osteoclast differentiation from monocyte / macrophage lineage cells involved in propagation and resolution of inflammatory bone destruction . Protein inhibitor of activated P40763 REA ( Q9Y6X2 REA ) was initially identified as a molecule that inhibits DNA binding of P40763 REA and regulates many transcription factors through distinct mechanisms . To analyze Q9Y6X2 REA function in osteoclasts in vivo , we have generated transgenic mice in which Q9Y6X2 REA is specifically expressed in the osteoclast lineage using the tartrate-resistant acid phosphatase ( TRAP ) gene promoter . Q9Y6X2 REA transgenic mice showed an osteopetrotic phenotype due to impairment of osteoclast differentiation . Overexpression of Q9Y6X2 REA in RAW 264.7 cells suppressed O14788 REA - induced osteoclastogenesis by inhibiting the expression of c-Fos and O95644 REA . Interestingly , Q9Y6X2 REA inhibits the transcriptional activity of microphthalmia-associated transcription factor ( O75030 REA ) independent of sumoylation . Down-regulation of Q9Y6X2 REA markedly enhances O14788 REA - mediated osteoclastogenesis in RAW 264.7 cells . Furthermore , overexpression of Q9Y6X2 REA in mouse primary osteoblast ( DB00925 ) , down-regulates O14788 REA expression induced by interleukin - 6 ( P05231 REA ) cytokine family , and inhibits osteoclast formation from bone marrow macrophages ( BMMs ) in vitro coculture system . Down-regulation of Q9Y6X2 REA leads to the accelerated expression of O14788 REA in DB00925 stimulated with P05231 REA and soluble P05231 REA receptor ( sIL - 6R ) . Taken together , our results clearly indicate that Q9Y6X2 REA negatively regulates O14788 REA - mediated osteoclastogenesis directly in osteoclast precursors and indirectly via osteoblasts .

DB00338 MEN , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 REA trafficking . DB00338 MEN is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 REA , a P-type H + / K + ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 MEN topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 MEN had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel 17 , or O75030 REA mRNA levels . Although melanocytes do not express P20648 REA , they do express Q04656 REA , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 REA relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 MEN treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 REA and by enhancing degradation of tyrosinase .

DB01109 MEN ' s anti-inflammatory effects require glucosamine 6 - O-sulfation and are mediated by blockade of L - and P-selectins . DB01109 MEN has been used clinically as an anticoagulant and antithrombotic agent for over 60 years . Here we show that the potent anti-inflammatory property of heparin results primarily from blockade of P16109 REA and P14151 REA . DB01109 MEN and chemically modified analogs were tested as inhibitors of selectin binding to immobilized sialyl Lewis ( X ) and of cell adhesion to immobilized selectins or thrombin-activated endothelial cells . Compared with unfractionated heparin , the modified heparinoids had inhibitory activity in this general order : over-O-sulfated heparin > heparin > 2 - O , 3 - O-desulfated > or = N-desulfated / N-acetylated heparin > or = carboxyl-reduced heparin > or = N - , 2 - O , 3 - O-desulfated heparin > > 6 - O-desulfated heparin . The heparinoids also showed similar differences in their ability to inhibit thioglycollate-induced peritonitis and oxazolone-induced delayed-type hypersensitivity . Mice deficient in P - or L-selectins showed impaired inflammation , which could be further reduced by heparin . However , heparin had no additional effect in mice deficient in both P - and L-selectins . We conclude that ( a ) heparin ' s anti-inflammatory effects are mainly mediated by blocking P - and P14151 REA - initiated cell adhesion ; ( b ) the sulfate groups at P13671 REA on the glucosamine residues play a critical role in selectin inhibition ; and ( c ) some non-anticoagulant forms of heparin retain anti-inflammatory activity . Such analogs may prove useful as therapeutically effective inhibitors of inflammation .

Therapy with interferon-beta modulates endogenous catecholamines in lymphocytes of patients with multiple sclerosis . OBJECTIVE : To investigate the endogenous dopaminergic / adrenergic system of lymphocytes in multiple sclerosis ( MS ) patients during treatment with interferon ( IFN ) - beta . METHODS : Patients with relapsing-remitting MS undergoing IFN-beta treatment were prospectively studied during the first year of treatment . Circulating lymphocytes were obtained at baseline and after 1 , 3 , 6 and 12 months of treatment and assayed for catecholamine ( CA ) production and mRNA expression of tyrosine hydroxylase ( TH , the rate-limiting enzyme in the synthesis of CA ) , beta ( 2 ) - adrenoceptors ( AR ) and D2 , D3 and D5 dopaminergic receptors ( DR ) . RESULTS : In cells from patients treated with IFN-beta for 12 months the production of CA hugely increased and was less sensitive to P01579 REA - induced inhibition . Expression of mRNA for TH , beta ( 2 ) - AR and P21918 REA was already enhanced after 1 month and further increased up to 6-12 months of treatment . On the contrary , P14416 REA mRNA progressively decreased and P35462 REA mRNA did not significantly change over the whole study period . CONCLUSIONS : In MS patients IFN-beta treatment enhances the ability of lymphocytes to produce CA , and induces extensive modifications of both beta ( 2 ) - AR and DR-operated pathways . The clinical relevance of these effects deserves consideration .

P04035 REA inhibitors up-regulate anti-aging klotho mRNA via RhoA inactivation in IMCD 3 cells . OBJECTIVE : Q9UEF7 is thought to play a critical role in the development of age-related disorders including arteriosclerosis . Statins may exert vascular protective effects , independent of the lowering of plasma cholesterol levels . We investigated the impact of statins on mRNA expression of the age-suppressor gene , klotho in mIMCD 3 cells . METHODS AND RESULTS : Q9UEF7 mRNA levels were evaluated with real-time RT-PCR . DB01076 MEN and pitavastatin increased the expression of klotho mRNA in a dose-dependent manner . This stimulatory effect was abolished by the addition of mevalonate , GGPP and FPP , essential molecules for isoprenylation of the small GTPase Rho . As was the case with the statin treatment , inhibition of Rho-kinase by Y27632 up-regulated klotho mRNA . In contrast to the statin treatment , stimulation with angiotensin II down-regulated klotho mRNA expression without obvious morphological changes . Furthermore , pretreatment with atorvastatin blunted the angiotensin II-induced response and ameliorated the decrease in klotho mRNA expression towards basal levels . RhoA activity was further evaluated by detection of its translocation . Angiotensin II activated RhoA , whereas statins potently inactivated RhoA and blocked RhoA activation by angiotensin II . CONCLUSION : Statins inactivate the RhoA pathway , resulting in over-expression of klotho mRNA , which may contribute to the novel pleiotropic effects of statins towards vascular protection .

Dopamine agonists upregulate P05231 REA and P10145 REA production in human keratinocytes . AIM : Catecholamines regulate functions of the nervous , neuroendocrine and immune systems . Dopamine may modulate the activity of keratinocytes , which play a role in secreting cytokines and chemokines . The aim of this study was to evaluate the effect of dopaminergic agonists on the production of P05231 REA and P10145 REA by a non-tumoral human keratinocyte cell line ( HaCaT ) . METHODS : Cells were stimulated with dopamine and the P14416 REA agonist cabergoline . Levels of P05231 REA and P10145 REA in culture supernatants were then determined . Cell proliferation was also assessed . Assays were carried out in the presence or absence of the dopaminergic and β-adrenergic receptor antagonists ( sulpiride and propranolol , respectively ) and ascorbic acid . RESULTS : Dopamine stimulated the production of P05231 REA and P10145 REA in a concentration-dependent manner . The effects observed on the secretion of P05231 REA were more potent than those corresponding to P10145 REA and were reduced by ascorbic acid . The dopamine-induced P05231 REA secretion was partially reduced by sulpiride and abrogated by propranolol . The latter drug was able to block the effect of dopamine on the secretion of P10145 REA . The cabergoline-induced P05231 REA release was reduced by sulpiride . Cell viability was not affected by any of the drugs . CONCLUSIONS : Dopaminergic agonists can stimulate keratinocytes to produce P05231 REA and P10145 REA which are related to inflammatory cutaneous processes . These effects are mediated by dopaminergic and β-adrenergic receptors and by receptor-independent oxidative mechanisms .

Solution structure of APETx 1 from the sea anemone Anthopleura elegantissima : a new fold for an Q12809 REA toxin . APETx 1 is a 42 - amino acid toxin purified from the venom of the sea anemone Anthopleura elegantissima . This cysteine-rich peptide possesses three disulfide bridges ( C4 - C37 , P13671 REA - C30 , and C20 - C38 ) . Its pharmacological target is the Ether-a-gogo potassium channel . We herein determine the solution structure of APETx 1 by use of conventional two-dimensional 1H - NMR techniques followed by torsion angle dynamics and refinement protocols . The calculated structure of APETx 1 belongs to the disulfide-rich all-beta structural family , in which a three-stranded anti-parallel beta-sheet is the only secondary structure . APETx 1 is the first Ether-a-gogo effector discovered to fold in this way . We therefore compare the structure of APETx 1 to those of the two other known effectors of the Ether-a-gogo potassium channel , CnErg 1 and BeKm - 1 , and analyze the topological disposition of key functional residues proposed by analysis of the electrostatic anisotropy . The interacting surface is made of a patch of aromatic residues ( Y5 , Y32 , and F33 ) together with two basic residues ( K8 and P05783 REA ) at the periphery of the surface . We pinpoint the absence of the central lysine present in the functional surface of the two other Ether-a-gogo effectors .

Effective dasatinib uptake may occur without human organic cation transporter 1 ( O15245 REA ) : implications for the treatment of imatinib-resistant chronic myeloid leukemia . We have previously shown that imatinib uptake into chronic myeloid leukemia ( CML ) cells is dependent on human organic cation transporter 1 ( O15245 REA ; O15245 REA ) , and that low O15245 REA expression is an important determinant of clinical outcome to imatinib treatment . We hypothesized that dasatinib might be transported differently than imatinib , possibly accounting for its favorable effects in imatinib-resistant patients . ( 14 ) C-dasatinib uptake was greater in KCL 22 - transfected cells with pcDNA 3 - O15245 REA plasmid ( high O15245 REA - expressing cells ) than in control cells ( P = . 02 ) . However , hOCT inhibitors did not decrease dasatinib uptake into either control or primary cells , in contrast to their block on imatinib uptake . Dasa-tinib decreased the level of phosphorylated CrkL to 49.9 % in control and 40.3 % in high O15245 REA - expressing cells . Dasa-tinib efflux was investigated in confluent P08183 REA - transfected MDCKII cell monolayers . Both dasatinib and imatinib were transported from the basal to the apical layer , indicating that they were transported by P08183 REA , which was confirmed using the P08183 REA inhibitor PSC 833 ( P = . 001 and P < . 001 , respectively ) . Compared with imatinib , dasatinib achieved superior intracellular levels and P11274 REA - P00519 REA suppression even in cells with low or blocked O15245 REA . Efflux of dasatinib and imatinib appear similar via P08183 REA . Dasatinib may therefore offer an advantage over imatinib in patients with low O15245 REA expression .

[ DB00391 MEN in the management of functional dyspepsia and delayed gastric emptying ] . DB00391 MEN is a sulpiride isomer that exerts its prokinetic action through a dual mechanism : 1 ) as a P14416 REA antagonist and 2 ) as a serotonin 5HT ( 4 ) receptor agonist , conferring this drug with a cholinergic effect . At a dosage of 25mg three times daily , levosulpiride accelerates gastric and gallbladder emptying . Clinical trials have shown that this agent is more effective than placebo in reducing the symptoms of dyspepsia , while comparative studies have demonstrated that its effect is similar or superior to that of other dopamine antagonists . The safety profile of levosulpiride is good and the frequency of adverse events is similar to that of other D ( 2 ) dopamine antagonists . Therefore , this drug is a useful therapeutic option in the management of patients with functional dyspepsia , as well as in those with delayed gastric emptying .

DB01296 MEN sulfate inhibits P01375 REA and P01579 REA - induced production of P05362 REA in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 MEN sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) - 1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE - 19 cells were used as a model to determine the effects of GS on the expression of the P05362 REA gene upregulated by P01375 REA or P01579 REA , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 REA were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 REA and P01579 REA increased the expression of P05362 REA at the mRNA and protein levels in a time - and dose-dependent manner in ARPE - 19 cells . GS effectively downregulated the P01375 REA - or P01579 REA - induced expression of P05362 REA in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 REA and phosphorylated P42224 REA in P01579 REA - stimulated ARPE - 19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 REA gene in ARPE - 19 cell stimulated with P01375 REA or P01579 REA through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 REA . This study has demonstrated a potentially important property of GS in reducing P05362 REA mediated inflammatory mechanisms in the eye .

A genomic reservoir for Tnfrsf genes is developmentally regulated and imprinted in the mouse . P01375 REA receptor superfamily is composed of at least 26 members in the mouse , three of which exist as a cluster within the imprinted Kcnq 1 domain on chromosome 7 . Tnfrsf 22 , 23 and 26 contain typical cystein-rich domains and Tnfrsf 22 and 23 can bind ligands but have no signaling capacity . Thus , they are assumed to be decoy receptors . The developmental expression profile of these genes is unknown and knowledge of their imprinting patterns is incomplete and controversial . We found that all three genes are expressed during mouse embryonic development , and that they have a strong maternal bias , indicating that they may be affected by the KvDMR , the Kcnq 1 imprinting control region . We found expression of an antisense non-coding RNA , AK155734 , in embryos and some neonatal tissues . This RNA overlaps the Tnfrsf 22 and possibly the Tnfrsf 23 coding regions and is also expressed with a maternal bias . We were interested in exploring the evolutionary origins of the three Tnfrsf genes , because they are absent in the orthologous human Kcnq 1 domain . To determine whether the genes were deleted from humans or acquired in the rodent lineage , we performed phylogenetic analyses . Our data suggest that TNFRSF sequences were duplicated and / or degenerated or eliminated from the P51787 REA region several times during the evolution of mammals . In humans , multiple mutations ( point mutations and / or deletions ) have accumulated on the ancestral TNFRSF , leaving a single short non-functional sequence .

Imatinib has the potential to exert its antileukemia effects by down-regulating Q12809 REA K + channels in chronic myelogenous leukemia . Imatinib is a powerful protein tyrosine kinase ( PTK ) inhibitor that specifically targets P11274 REA - P00519 REA , P10721 REA , and P09619 REA kinases , has become the current first-line therapy for all newly diagnosed chronic myeloid leukemia ( CML ) . Beside PTKs , PTK inhibitors alter the activity of a large number of voltage-dependent ion channels . Q12809 REA K ( + ) channels are highly expressed in leukemia cells and appear of exceptional importance in favoring leukemogenesis . The present study explored a possible regulatory effect of imatinib upon Q12809 REA K ( + ) channels as a means to uncover new molecular events involved in the antileukemic activity of this PTK inhibitor in CML . The results demonstrated that Q12809 REA was highly detected in K562 cells and primary CML cells , and down-regulated by imatinib at mRNA and protein levels . Furthermore , imatinib markedly reduced hERG currents in HEK 293T - hERG cells , this effect was accompanied by inhibition of CML cell proliferation and apoptosis , as well as suppression of vascular endothelial growth factor ( P15692 REA ) secretion . Moreover , these antileukemia effects of imatinib were potentiated by E - 4031 , a specific Q12809 REA inhibitor . Together , these results provide evidence of a novel potential molecular mechanism of antileukemic activities by imatinib which , independent of targeting tyrosine kinase , highlight Q12809 REA K ( + ) channels as a therapeutic target for CML treatment .

Activation of PPARgamma by rosiglitazone attenuates intestinal Cl - secretion . The thiazolidinedione ( TZD ) drugs rosiglitazone ( Ro ) and pioglitazone ( Po ) are PPARgamma agonists in widespread clinical use as insulin-sensitizing agents in Type 2 diabetes . On the basis of recent evidence implicating PPARgamma as a positive modulator of intestinal epithelial differentiation , we hypothesized that TZD drugs might attenuate intestinal secretory function . To evaluate this possibility , we examined the effects of Ro and Po on electrogenic Cl - secretion [ short-circuit current ( I ( sc ) ) ] in mouse intestinal segments and in cultured human intestinal epithelial cells ( HT29 - Cl . 19A ) . As hypothesized , oral administration of Ro ( 20 mg.kg ( - 1 ) . day ( - 1 ) ) to mice for 8 days markedly reduced intestinal I ( sc ) responses to DB02527 ( forskolin ) - and Ca2 + ( carbachol ) - dependent stimuli . In these Ro-treated mice , cholera toxin-induced intestinal fluid accumulation was reduced 65 % . With continued Ro treatment , the I ( sc ) response to carbachol recovered significantly , whereas that to forskolin remained attenuated . Treatment of HT29 cells for 5 days with 10 muM Ro or Po in vitro brought about a similar hyposecretory state . In HT29 cells , the loss of DB02527 - dependent Cl - secretion was attributable to a reduced expression of P13569 REA Cl - channel , P51787 REA K + channel , and Na-K - 2Cl cotransporter - 1 proteins . The transient loss of Ca2 + - dependent Cl - secretion involved an impairment of basolateral Ca2 + - stimulated K + channel activity without a detectable loss of K ( Ca ) 3.1 channel protein . Our results establish TZD drugs as important modulators of intestinal Cl - secretory function .

DB09280 - DB08820 MENMAX DB08820 MEN in Patients with Cystic Fibrosis Homozygous for Phe 508del P13569 REA .

Upregulation of P51787 REA / P15382 REA K + channels by Q9UEF7 . Q9UEF7 is a transmembrane protein expressed primarily in kidney , parathyroid gland , and choroid plexus . The extracellular domain could be cleaved off and released into the systemic circulation . Q9UEF7 is in part effective as β-glucuronidase regulating protein stability in the cell membrane . Q9UEF7 is a major determinant of aging and life span . Overexpression of Q9UEF7 increases and Q9UEF7 deficiency decreases life span . Q9UEF7 deficiency may further result in hearing loss and cardiac arrhythmia . The present study explored whether Q9UEF7 modifies activity and protein abundance of P51787 REA / P15382 REA , a K ( + ) channel required for proper hearing and cardiac repolarization . To this end , cRNA encoding P51787 REA / P15382 REA was injected in Xenopus oocytes with or without additional injection of cRNA encoding Q9UEF7 . P51787 REA / P15382 REA expressing oocytes were treated with human recombinant Q9UEF7 protein ( 30 ng / mL ) for 24 h . Moreover , oocytes which express both P51787 REA / P15382 REA and Q9UEF7 were treated with 10 μM DSA L ( D-saccharic acid -1,4- lactone ) , a β-glucuronidase inhibitor . The P51787 REA / P15382 REA depolarization-induced current ( I ( Ks ) ) was determined utilizing dual electrode voltage clamp , while P51787 REA / P15382 REA protein abundance in the cell membrane was visualized utilizing specific antibody binding and quantified by chemiluminescence . P51787 REA / P15382 REA channel activity and P51787 REA / P15382 REA protein abundance were upregulated by coexpression of Q9UEF7 . The effect was mimicked by treatment with human recombinant Q9UEF7 protein ( 30 ng / mL ) and inhibited by DSA L ( 10 μM ) . In conclusion , Q9UEF7 upregulates P51787 REA / P15382 REA channel activity by “ mainly ” enhancing channel protein abundance in the plasma cell membrane , an effect at least partially mediated through the β-glucuronidase activity of Q9UEF7 protein .

Augmented potassium current is a shared phenotype for two genetic defects associated with familial atrial fibrillation . Mutations in multiple genes have been implicated in familial atrial fibrillation ( AF ) , but the underlying mechanisms , and thus implications for therapy , remain ill-defined . Among 231 participants in the Vanderbilt AF Registry , we found a mutation in P51787 REA ( encoding the alpha-subunit of slow delayed rectifier potassium current [ I ( Ks ) ] ) and separately a mutation in natriuretic peptide precursor A ( P01160 REA ) gene ( encoding atrial natriuretic peptide , P01160 REA ) , both segregating with early onset lone AF in different kindreds . The functional effects of these mutations yielded strikingly similar I ( Ks ) " gain-of-function . " In Chinese Hamster Ovary ( CHO ) cells , coexpression of mutant P51787 REA with its ancillary subunit P15382 REA generated approximately 3 - fold larger currents that activated much faster than wild-type ( WT ) - I ( Ks ) . Application of the WT P01160 REA peptide fragment produced similar changes in WT-I ( Ks ) , and these were exaggerated with the mutant P01160 REA S64R peptide fragment . Anantin , a competitive P01160 REA receptor antagonist , completely inhibited the changes in I ( Ks ) gating observed with P01160 REA S64R . Computational simulations identified accelerated transitions into open states as the mechanism for variant I ( Ks ) gating . Incorporating these I ( Ks ) changes into computed human atrial action potentials ( AP ) resulted in 37 % shortening ( 120 vs . 192 ms at 300 ms cycle length ) , reflecting loss of the phase II dome which is dependent on L-type calcium channel current . We found striking functional similarities due to mutations in P51787 REA and P01160 REA genes which led to I ( Ks ) " gain-of-function " , atrial AP shortening , and consequently altered calcium current as a common mechanism between diverse familial AF syndromes .

DB00588 MEN - induced regulation of the balance within macrophage subpopulations . In asthma , treatment with inhaled corticosteroids reduces chronic peribronchial inflammation and restores the balance within macrophage subpopulations . This study investigates whether corticosteroids can regulate monocyte differentiation in vitro and thereby influence the balance of functionally distinct macrophages . Graded doses of fluticasone propionate ( FP ) were added to cultures of normal peripheral blood monocytes in the presence or absence of P05112 REA . Cells were harvested after 7 days ' culture . Double immunofluorescence studies were performed on cytospins of differentiated macrophages using the MoAbs RFD 1 and RFD 7 to distinguish inductive and suppressive macrophages by their respective phenotypes . Macrophage function was determined by quantifying allostimulation in a mixed leucocyte reaction and by measuring tumour necrosis factor-alpha ( P01375 REA ) production . FP reduced the number of mature cells with a D1 + antigen-presenting phenotype and up-regulated the development of cells with the D1 / D7 + and D7 + phenotypes . Functionally , this was associated with reduced stimulation of T cell proliferation in a mixed leucocyte reaction ( P08235 REA ) . DB00588 MEN also reversed the increase in both D1 + expression and P01375 REA production induced by P05112 REA . The effect of FP persisted for 24 h after removal of FP from the culture medium . These results suggest that FP treatment of asthmatics may have a direct beneficial effect by normalizing the macrophage subset imbalance that contributes to the chronic peribronchial inflammation present in this condition .

Exposure to an organophosphate ( DB00677 MEN ) during a defined period in neonatal life induces permanent changes in brain muscarinic receptors and behaviour in adult mice . The organophosphate DB00677 MEN ( DB00677 MEN ) is a well-known inhibitor of cholinesterases . We have recently observed that neonatal exposure to a single subsymptomal dose of DB00677 MEN induces permanent alterations in muscarinic cholinergic receptors ( MAChRs ) and in spontaneous behaviour , in the mice as adults . In order to determine if there is a critical period for these effects , neonatal mice were given a single oral dose of 1.5 mg / kg DB00677 MEN b.wt . on postnatal day 3 , 10 or 19 , causing equal inhibition of P22303 REA . At the adult age of 4 months the mice were tested for spontaneous motor behaviour , and were subsequently sacrificed for measurement of density of MAChRs and subpopulations of MAChRs in the cerebral cortex by using the antagonist quinuclidinyl benzilate ( [ 3H ] QNB ) , and agonist carbachol , respectively . At adult age , mice exposed to DB00677 MEN on postnatal day ( P01160 REA ) 3 or 10 showed significant ( P < or = 0.01 ) alterations in spontaneous motor behaviour and a significant ( P < or = 0.01 ) decrease in muscarinic receptor density . There were no alterations mice exposed on P01160 REA 19 . The proportions and affinity-constants of high - and low-affinity MAChR binding sites were not affected in mice showing altered MAChR density . The lack of effect on mice exposed on P01160 REA 19 was not due to differences in P22303 REA activity .

DB01184 SUB treatment for gastroparesis : demographic and pharmacogenetic characterization of clinical efficacy and side-effects . BACKGROUND : DB01184 SUB is a useful alternative to metoclopramide for treatment of gastroparesis due to better tolerability . Effectiveness and side-effects from domperidone may be influenced by patient-related factors including polymorphisms in genes encoding drug-metabolizing enzymes , drug transporters , and domperidone targets . AIMS : The aim of this study was to determine if demographic and pharmacogenetic parameters of patients receiving domperidone are associated with response to treatment or side-effects . METHODS : Patients treated with domperidone for gastroparesis provided saliva samples from which DNA was extracted . Fourteen single-nucleotide polymorphisms ( SNPs ) in seven candidate genes ( P08183 REA , P10635 REA , P14416 REA , P15382 REA , Q9Y6J6 REA , Q12809 REA , P51787 REA ) were used for genotyping . SNP microarrays were used to assess single-nucleotide polymorphisms in the ADRA 1A , P35368 REA , and P25100 REA loci . RESULTS : Forty-eight patients treated with domperidone participated in the study . DNA was successfully obtained from each patient . Age was associated with effectiveness of domperidone ( p= 0.0088 ) . Genetic polymorphism in Q12809 REA was associated with effectiveness of domperidone ( p= 0.041 ) . The efficacious dose was associated with polymorphism in P08183 REA gene ( p= 0.0277 ) . The side-effects of domperidone were significantly associated with the SNPs in the promoter region of P25100 REA gene . CONCLUSIONS : Genetic characteristics associated with response to domperidone therapy included polymorphisms in the drug transporter gene P08183 REA , the potassium channel Q12809 REA gene , and α1D - - adrenoceptor P25100 REA gene . Age was associated with a beneficial response to domperidone . If verified in a larger population , this information might be used to help determine which patients with gastroparesis might respond to domperidone and avoid treatment in those who might develop side-effects .

Enhancement of cytotoxicity of natural product drugs against multidrug resistant variant cell lines of human head and neck squamous cell carcinoma and breast carcinoma by tesmilifene . N , N-diethyl - 2 - [ 4 - ( phenylmethyl ) phenoxyl ] ethanamine ( tesmilifene ) , a tamoxifen derivative with antihistamine activity , greatly enhanced the survival of doxorubicin-treated , advanced stage breast cancer patients in a phase III trial . However , the molecular basis of tesmilifene action is not firmly established . The effects of tesmilifene on activity of several anticancer drugs was investigated using human head and neck squamous cell carcinoma ( HNSCC ) and breast carcinoma cell lines as a model system . Multidrug resistant ( MDR ) variants of an HNSCC cell line , HN - 5a / V15e , and a breast carcinoma cell line , MCF - 7 / V25a , both highly overexpressed mdr 1 ( P08183 REA ) mRNA and the proteins P-glycoprotein and glutathione transferase-pi . Drug sensitivities were measured by a vital stain after 4 days of continuous exposure to anticancer drug in the absence and presence of tesmilifene at a concentration that alone had no antiproliferative effect . DB04905 MEN had minimal effect on drug cytotoxicity against the parental cell lines . However , the same tesmilifene treatment enhanced cytotoxicity of docetaxel , paclitaxel , epirubicin , doxorubicin , and vinorelbine against both MDR cell lines by up to 50 % . Flow cytometric measurement of annexin V / propidium iodide staining demonstrated that tesmilifene increased the killing of HN - 5a / V15e cells caused by docetaxel after 24 and 48h exposure . DB04905 MEN increased accumulation of radiolabelled vincristine in HN - 5a / V15e cells , over 4h , by up to 100 % . The results suggest that tesmilifene might be effective in the treatment of tumors that are resistant to natural product drugs . The mechanism of enhancement appears to be related to expression of an ABC pump-dependent , MDR phenotype .