interacts_with DB00707?
P40189 -linked signal transduction promotes the differentiation and maturation of dendritic cells . In order to explore the role of P40189 -linked signal transduction in the differentiation and maturation of dendritic cells ( DC ) , the mAb , B- P28222 , an agonist of P40189 , was used for the activation of P40189 on DC . The effects of cytokines and of anti- P40189 mAb on the proliferation of DC , and their expression of IL-12 and P33681 ( P33681 -1 ) by DC were evaluated . DC differentiating from peripheral blood mononuclear cells did not express the P05231 receptor alpha chain , but expressed P40189 . Anti- P40189 mAb promoted the proliferation of DC , induced by P05112 and granulocyte macrophage colony stimulating factor ( GM- P04141 ) , by up-regulating the GM- P04141 receptor on DC . DC induced by P40189 mAb and cytokines expressed DC-derived CC chemokine , as measured by RT-PCR . Induced DC also stimulated strong proliferation of autologous T cells in mixed lymphocyte reaction since an up-regulated expression of IL-12 and P33681 ( P33681 -1 ) was observed in DC activated by anti- P40189 mAb . Thus , P40189 signal transduction is important for the differentiation and maturation of DC . P08235 antagonizes Dot1a-Af9 complex to increase αENaC transcription . DB04630 is a major regulator of Na(+) absorption and acts by activating the mineralocorticoid receptor ( MR ) to stimulate the epithelial Na(+) channel ( ENaC ) . MR(-/-) mice exhibited pseudohypoaldosteronism type 1 ( hyponatremia , hyperkalemia , salt wasting , and high levels of aldosterone ) and died around postnatal day 10 . However , if and how MR regulates ENaC transcription remain incompletely understood . Our earlier work demonstrated that aldosterone activates αENaC transcription by reducing expression of Dot1a and Af9 and by impairing Dot1a-Af9 interaction . Most recently , we reported identification of a major Af9 binding site in the αENaC promoter and upregulation of αENaC mRNA expression in mouse kidneys lacking Dot1a . Despite these findings , the putative antagonism between the MR/aldosterone and Dot1a-Af9 complexes has never been addressed . The molecular defects leading to PHA-1 in MR(-/-) mice remain elusive . Here , we report that MR competes with Dot1a to bind Af9 . MR/aldosterone and Dot1a-Af9 complexes mutually counterbalance ENaC mRNA expression in inner medullary collecting duct 3 ( IMCD3 ) cells . Real-time RT-quantitative PCR revealed that 5-day-old MR(-/-) vs. MR(+/+) mice had significantly lower αENaC mRNA levels . This change was associated with an increased Af9 binding and H3 Q5XKE5 hypermethylation in the αENaC promoter . Therefore , this study identified MR as a novel binding partner and regulator of Af9 and a novel mechanism coupling MR-mediated activation with relief of Dot1a-Af9-mediated repression via MR-Af9 interaction . Impaired ENaC expression due to failure to inhibit Dot1a-Af9 may play an important role in the early stages of PHA-1 ( before postnatal day 8 ) in MR(-/-) mice . DB00998 . black triangle DB00998 , a new serotonin receptor agonist developed for the acute treatment of migraine , has high affinity for serotonin P28222 and P28221 receptor subtypes and is a potent stimulator of contraction in human basilar arteries. black triangle A long terminal elimination half-life ( approximately 26 hours ) is a distinctive pharmacokinetic feature of frovatriptan which appears to be independent of dose , age , gender and renal function. black triangle A single oral dose of frovatriptan 2.5mg was effective in the acute treatment of migraine providing meaningful relief within 2 hours to approximately twice as many recipients as placebo in clinical trials. black triangle Consistent relief of migraine symptoms was achieved in patients who treated a number of consecutive attacks with frovatriptan and the incidence of 24-hour migraine recurrence was reduced. black triangle DB00998 was well tolerated in clinical trials , with the overall incidence of adverse events occurring with frovatriptan 2.5mg only slightly higher than that reported with placebo . Mild to moderate fatigue , nausea and paraesthesia were the most commonly reported drug-related adverse events . P35354 promotes early atherosclerotic lesion formation in P01130 -deficient mice . BACKGROUND : Atherosclerosis has features of an inflammatory disease . Because cyclooxygenase ( P36551 ) -2 is expressed in atherosclerotic lesions and promotes inflammation , we tested the hypotheses that selective P35354 inhibition would reduce early lesion formation in P01130 -deficient ( P01130 -/- ) mice and that macrophage P35354 expression contributes to atherogenesis in P01130 -/- mice . METHODS AND RESULTS : Treatment of male P01130 -/- mice fed the Western diet with rofecoxib or indomethacin for 6 weeks resulted in significant reductions in atherosclerosis in the proximal aorta ( 25 % and 37 % ) and in the aorta en face ( 58 % and 57 % ) , respectively . DB00533 treatment did not inhibit platelet thromboxane production , a P23219 -mediated process , but it significantly reduced the urinary prostacyclin metabolite 2,3-dinor-6-keto-PGF1alpha . Fetal liver cell transplantation was used to generate P01130 -/- mice null for expression of the P35354 gene by macrophages . After 8 weeks on the Western diet , P35354 -/- --> P01130 -/- mice developed significantly less ( 33 % to 39 % ) atherosclerosis than control P35354 +/+ --> P01130 -/- mice . In both the inhibitor studies and the transplant studies , serum lipids did not differ significantly between groups . CONCLUSIONS : The present studies provide strong pharmacological and genetic evidence that P35354 promotes early atherosclerotic lesion formation in P01130 -/- mice in vivo . These results support the potential of anti-inflammatory approaches to the prevention of atherosclerosis . Lectin-like oxidized P01130 -1 ( P78380 ) acts as a receptor for remnant-like lipoprotein particles ( RLPs ) and mediates RLP-induced migration of vascular smooth muscle cells . OBJECTIVE : Remnant-like lipoprotein particles ( RLPs ) have been implicated in atherogenesis especially by diabetic dyslipidemia ; however , their receptor(s) and effects on vascular smooth muscle cells ( VSMCs ) remain unclear . In this study , we examined if lectin-like oxidized P01130 -1 ( P78380 ) acts as a receptor for RLPs and its biological effects in VSMCs . METHODS AND RESULTS : RLPs were isolated from human plasma by immunoaffinity gel containing anti-apolipoprotein A-I and anti-apolipoprotein B-100 monoclonal antibodies . DiI-labeled RLPs were taken up by CHO- P04264 cells stably expressing P78380 but not by wild-type CHO- P04264 cells . RLPs induced P78380 expression and cell migration in bovine VSMCs ( BVSMCs ) , which were significantly suppressed by transfection with P78380 specific siRNAs . Inhibitors of metalloproteinases , epidermal growth factor ( P01133 ) receptor tyrosine kinase , heparin-binding P01133 -like growth factor ( HB- P01133 ) , p38 mitogen-activated protein kinase ( p38 MAPK ) , MAPK kinase ( Q02750 ) and phosphoinositide 3-kinase ( PI3K ) significantly blocked RLP-induced P78380 expression and cell migration of BVSMCs . CONCLUSIONS : The present study provides direct evidence that P78380 is a novel receptor for RLPs in VSMCs . P78380 -mediated uptake of RLPs may thus play important roles in atherogenesis by inducing P78380 expression and VSMC migration especially in the settings of postprandial hyperlipidemia , diabetes and metabolic syndrome . Immunoglobulin treatment reduces atherosclerosis in apolipoprotein E-/- low-density lipoprotein receptor-/- mice via the complement system . Atherosclerosis is associated with activation of the immune system . Intravenously applied normal polyclonal immunoglobulins ( DB00028 ) have broad therapeutic applications in the treatment of autoimmune and systemic inflammatory diseases . Recently , DB00028 have been shown to inhibit atherogenesis in experimental animal models . To investigate the role of the complement system in this process , we used third complement component-deficient ( P01024 (-/-) ) and control atherosclerosis-prone apolipoprotein E ( ApoE ) and low-density lipoprotein receptor ( P01130 ) double knock-out mice fed a normal diet . DB00028 treatment reduced lesion fraction area in the aortic root of complement-sufficient mice whereas the lesion fraction area of P01024 (-/-) mice was not affected . Thus , complement activation plays a role in the anti-atherosclerotic effects of DB00028 , possibly by P01024 -derived fragments generated through Fc-dependent complement activation . Acute renal failure from hemoglobinuric and interstitial nephritis secondary to iodine and mefenamic acid . DB00784 ingestion , usually in excess and over prolonged period is known to produce interstitial nephritis , or less commonly papillary necrosis , with acute renal failure . However , it is not dose-dependent for the induction of tubulointerstitial damage . Excess iodine ingestion is known to produce toxicity and possible death , but acute renal failure is rare . There is evidence from clinical and experimental data that iodine has toxic effect on tubular epithelial cells . Iodine has not been documented to produce red cell hemolysis and hemoglobinuria . We present a unique case of acute renal failure from hemoglobinuric and acute interstitial nephritis secondary to suicidal ingestion of potassium iodide solution and also ingestion of a few mefenamic acid tablets . These agents led to potentiation of the renal injury from hemoglobinuric tubulopathy , probably from the iodine , and renal dysfunction from alteration of renal perfusion by selective P23219 inhibition of prostaglandin production , and induction of acute interstitial nephritis from mefenamic acid , leading to acute renal failure which was reversible by hemodialysis and supportive therapy . Zerumbone suppresses phorbol ester-induced expression of multiple scavenger receptor genes in THP-1 human monocytic cells . Unregulated uptake of oxidized low-density lipoproteins ( ox-LDL ) via macrophage scavenger receptors ( SRs ) , such as lectin-like ox- P01130 -1 ( P78380 ) , is a key event in atherosclerosis . In the present study , we used differentiated Caco-2 cells as a model of the human small intestine to evaluate the suppressive effects of 16 traditional food items selected from Okinawa on 12-O-tetradecanoylphorbol-13-acetate ( TPA ) -induced P78380 mRNA expression in THP-1 human monocyte-like cells . Three Zingiberaceae plants , Curcuma aromatica Salisbury , Curcuma longa L. , and Zingiber zerumbet Smith , markedly suppressed that expression . When added to the apical sides of Caco-2 monolayers , zerumbone , a sesquiterpene from Z. zerumbet Smith , was found to permeate into the basolateral medium as an intact structure in a time-dependent manner . alpha-Humulene , a structural analog of zerumbone lacking the alpha,beta-unsaturated carbonyl group , did not suppress P78380 mRNA expression , indicating that its electrophilic moiety might play pivotal roles in its activities . Further , zerumbone attenuated the expression of SR-A , Q9H2A7 , and P16671 , but not that of P34810 or Q8WTV0 , leading to a blockade of DiI-acLDL uptake , while it also inhibited the transcriptional activities of activator protein-1 and nuclear factor-kappaB . Together , our results indicate that zerumbone is a potential phytochemical for regulating atherosclerosis with reasonable action mechanisms . Binding of low density lipoprotein to platelet apolipoprotein E receptor 2 ' results in phosphorylation of p38MAPK . Binding of low density lipoprotein ( LDL ) to platelets enhances platelet responsiveness to various aggregation-inducing agents . However , the identity of the platelet surface receptor for LDL is unknown . We have previously reported that binding of the LDL component apolipoprotein B100 to platelets induces rapid phosphorylation of p38 mitogen-activated protein kinase ( p38MAPK ) . Here , we show that LDL-dependent activation of this kinase is inhibited by receptor-associated protein ( P30533 ) , an inhibitor of members of the P01130 family . Confocal microscopy revealed a high degree of co-localization of LDL and a splice variant of the P01130 family member apolipoprotein E receptor-2 ( apoER2 ' ) at the platelet surface , suggesting that apoER2 ' may contribute to LDL-induced platelet signaling . Indeed , LDL was unable to induce p38MAPK activation in platelets of apoER2-deficient mice . Furthermore , LDL bound efficiently to soluble apoER2 ' , and the transient LDL-induced activation of p38MAPK was mimicked by an anti-apoER2 antibody . Association of LDL to platelets resulted in tyrosine phosphorylation of apoER2 ' , a process that was inhibited in the presence of P50391 , an inhibitor of Src-like tyrosine kinases . Moreover , phosphorylated but not native apoER2 ' co-precipitated with the Src family member Fgr . This suggests that exposure of platelets to LDL induces association of apoER2 ' to Fgr , a kinase that is able to activate p38MAPK . In conclusion , our data indicate that apoER2 ' contributes to LDL-dependent sensitization of platelets . Glioma cell activation by Alzheimer 's peptide Abeta1-42 , alpha1-antichymotrypsin , and their mixture . We compared the effects ofAlzheimer 's peptide ( Abeta1-42 ) , a,-antichymotrypsin ( ACT ) and an ACT/Abeta1-42 mixture on human glioma DK-MG cells . The solution of Abeta ( 5 microM ) formed by 2-h incubation at room temperature induced tumour necrosis factor-alpha ( P01375 ) and interleukin ( IL ) -6 levels by 55 and 45 % , respectively , and increased gelatinase B activity by 67 % , while exposure of cells to the ACT/Abeta1-42 mixture ( 1:10 molar ratio ACT : Abeta1-42 ) under the same experimental conditions showed no effect on P05231 levels or gelatinase B activity , but strongly induced P01375 ( by 190 % ) , compared to the controls . Stimulation of the cells with Abeta1-42 alone , but not with ACT , increased by about 20 % low-density lipoprotein ( LDL ) uptake and mRNA levels for P01130 and P04035 , while the ACT/Abeta1-42 mixture significantly increased LDL uptake ( by 50 % ) , up-regulated mRNA levels for P01130 and P04035 by 48 and 63 % , respectively , and increased lipid accumulation by about 20-fold . These data suggest a possible new role for Abeta in Alzheimer 's disease through its interaction with the inflammatory reactant , ACT . P35354 promotes early atherosclerotic lesion formation in ApoE-deficient and C57BL/6 mice . Cyclooxygenase ( P36551 ) 2 is expressed in atherosclerotic lesions . We have previously reported that selective inhibition of P35354 reduces early atherosclerosis in P01130 deficient mice . To examine the role of P35354 in atherosclerosis in other mouse models , we studied the effects of selective P35354 inhibition ( by rofecoxib and NS-398 ) and nonselective P36551 inhibition ( by indomethacin ) on early atherosclerotic lesion formation in apolipoprotein E-deficient ( apoE(-/-) ) mice . Selective P35354 and nonselective P36551 inhibition reduced atherosclerosis in female apoE(-/-) mice by 35-38 % and 38-51 % in the proximal and en face aortas , respectively . Next we investigated the role of macrophage P35354 by transplanting P35354 (-/-) fetal liver cells into C57BL/6 mice and challenging the mice with an atherogenic diet . Genetic deletion of P35354 from hematopoietic cells reduced atherosclerosis by 51 % . In addition , LPS activated P35354 (-/-) macrophages had decreased expression of monocyte chemoattractant protein-1 ( P13500 ) and tumor necrosis factor-alpha ( TNFalpha ) . The results demonstrate that selective inhibition of P35354 and elimination of P35354 from macrophages significantly reduces early atherosclerotic lesion formation in apoE-deficient and C57BL/6 mice . These results are compatible with P35354 expression by macrophages having a proatherogenic role , and support the potential of anti-inflammatory therapeutic approaches for atherosclerosis . Modification of Q9H2A7 functions by multi-point mutations of basic amino acid residues . Q9H2A7 can function as a scavenger receptor , a chemokine and an adhesion molecule , and it could be an interesting player in the formation of atherosclerotic lesions . Our previous studies demonstrated that basic amino acid residues in the chemokine domain of Q9H2A7 are critical for its functions . In this study the combinations of the key basic amino acids in the chemokine domain of Q9H2A7 have been identified . Five combinations of basic amino acid residues that may form conformational motif for Q9H2A7 functions were selected for multi-point mutants . The double mutants of K61AR62A , R76AK79A , R82AH85A , and treble mutants of R76AR78AK79A , R78AR82AH85A were successfully constructed by replacing the combinations of two or three basic amino acid residues with alanine . After successful expression of these mutants on the cells , the functional studies showed that the cells expressing R76AK79A and R82AH85A mutants significantly increased the activity of oxLDL uptake compared with that of wild-type Q9H2A7 . Meanwhile , the cells expressing R76AK79A mutant also dramatically enhanced the phagocytotic activity of Q9H2A7 . However , the cells expressing the construct of combination of R78A mutation in R76AK79A or R82AH85A could abolish these effects . More interestingly , the adhesive activities were remarkably down regulated in the cells expressing the multi-point mutants respectively . This study revealed that some conformational motifs of basic amino acid residues , especially R76 with Q5XKE5 in Q9H2A7 , may form a common functional motif for its critical functions . R78 in Q9H2A7 has the potential action to stabilize the function of oxLDL uptake and bacterial phagocytosis . The results obtained may provide new insight for the development of drug target of atherosclerosis . Dimerization effect of sucrose octasulfate on rat P05230 . Fibroblast growth factors ( FGFs ) constitute a family of at least 23 structurally related heparin-binding proteins that are involved in regulation of cell growth , survival , differentiation and migration . DB01901 ( SOS ) , a chemical analogue of heparin , has been demonstrated to activate FGF signalling pathways . The structure of rat P05230 crystallized in the presence of SOS has been determined at 2.2 A resolution . SOS-mediated dimerization of P05230 was observed , which was further supported by gel-filtration experiments . The major contributors to the sulfate-binding sites in rat P05230 are Lys113 , Lys118 , Arg122 and Lys128 . An arginine at position 116 is a consensus residue in mammalian FGF molecules ; however , it is a serine in rat P05230 . This difference may be important for SOS-mediated P05230 dimerization in rat . DB01109 -binding growth factor type one and platelet-derived growth factor are required for the optimal expression of cell surface low density lipoprotein receptor binding activity in human adult arterial smooth muscle cells . Purified heparin-binding growth factor-1 ( P05230 ) stimulated low density lipoprotein binding , internalization , and degradation in isolated human adult arterial smooth muscle cells . Exposure of quiescent cells to P05230 in serum-free , defined medium increased both low density lipoprotein ( LDL ) receptor activity and de novo cholesterol biosynthesis . Both events preceded the onset of DNA synthesis by 6 to 9 h . P05230 acted additively with platelet-derived growth factor ( PDGF ) to maximally stimulate cell surface P01130 binding activity and DNA synthesis in the smooth muscle cells . The presence of LDL was required for maximal mitogenic activity of P05230 and PDGF . In the presence of LDL , growth factor-stimulated , proliferating human smooth muscle cells accumulated cholesterol ester and triglycerides . The results suggest that P05230 , PDGF , and LDL act together to promote the maximal proliferation of smooth muscle cells in culture . Chronic exposure to the three growth promoters may contribute to the smooth muscle cell hyperplasia and lipid accumulation observed in atherosclerotic lesions . [ DB00707 sodium ( Photofrin-II ) ] . DB00707 sodium ( DB00707 ) is a photosensitizer which distributes selectively to tumor tissues , and causes tumor cell death by combination with light irradiation . Photodynamic therapy ( PDT ) by combination of porfimer sodium and laser was developed as a new cancer therapy . Tumor selectivity of porfimer sodium are based on the following reasons ; 1 ) high affinity for lipoprotein , especially , low density lipoprotein ( LDL ) , 2 ) elevation of P01130 activity in cancer tissue , and 3 ) lack or imcompleteness of lymphatic system in cancer tissue . DB00707 sodium is activated by laser irradiation at 630 nm , which can reacts with tissue oxygen to produce highly reactive excited siglet oxygen ( 1O2 ) . This highly reactive molecule is subsequently capable of killing tumor cells through oxidation of cellular component like mitochondrial enzymes . In addition , this highly reactive intermediate causes destruction of the tumor capillaries , which accelerates tumor cell death . The growth suppression or lethal damage to tumor cells by PDT of porfimer sodium and excimer dye laser were observed in experimental tumor models . In human clinical trials , the rates of complete response ( CR ) for roentgenographically occult lung cancer , stage I lung cancer , superficial esophageal cancer , superficial gastric cancer and carcinoma in situ or dysplasia of the cervix were 84.8 % , 50.0 % , 90.0 % , 87.5 % and 94.4 % , respectively . The major side effects were cutaneous symptoms e.g. photosensitivity , pigmentation , increasing GOT , GPT but these symptoms were not severe . PDT using porfimer sodium and excimer dye laser must be clinically useful for the treatment of inoperable early cancer or conservation of organ functions . 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 . DB01064 inhibits fibroblast growth factor-2-induced growth of renal epithelial cells . The signal transduction pathways modulating P09038 effects in renal tubular epithelial cells ( RTEc ) are not completely understood . Since the DB02527 and the mitogen-activated protein kinase ( MAPK ) pathways can modulate the growth of RTEc , we studied whether two DB02527 elevating agents , isoproterenol and 8-bromo- DB02527 , would modulate basic fibroblast growth factor ( P09038 ) induction of MAPK activity ( P28482 ) and cell proliferation in human renal proximal tubular epithelial cells ( RPTEc ) and Madin-Darby canine kidney cells ( MDCK clone EI1 ) . DB01064 , but not P09038 , stimulated DB02527 production in RPTEc and MDCKEI1 cells . P09038 , isoproterenol , and 8-bromo- DB02527 alone increased P28482 activity in both cell types . However , isoproterenol and 8-bromo- DB02527 partially inhibited the P09038 induction of P28482 activity , but only isoproterenol inhibited the proliferation of both cell types . PD098059 ( 25 microM ) , an inhibitor of MAPK kinase ( Q02750 /2 ) , blocked the P09038 mitogenic effects , but did not affect the 8-bromo- DB02527 -induced mitogenic effects in MDCKEI1 cells . These findings suggest that activation of P28482 is required but not sufficient for mitogenesis in RTEc . We conclude that isoproterenol inhibits the growth-promoting effects of P09038 in RTEc via MEK-dependent and -independent pathways . [ Potentiation of the photocytotoxic effect of photofrin II : synergistic action of verapamil and lovastatin ] . DB00707 ( P2 ) is at the present time the most used drug in the photochemotherapy of tumors . As previous studies from our group have demonstrated that P2 is taken up by cells mainly via the low density lipoprotein ( LDL ) receptor pathway , we tried to increase the amount of drug delivered to cells by enhancing the P01130 expression . For this purpose , we used hydroxy methyl glutaryl DB01992 ( HMG- DB01992 ) reductase inhibitors such as lovastatin . In the present work , we show that the calcium antagonist verapamil , which is currently used in human chemotherapy to overcome multidrug resistance , enhances in a synergistic manner the potentiating action of lovastatin on the photocytotoxic effect of P2 . Effects of mitiglinide ( S 21403 ) on Kir6.2/ Q09428 , Kir6.2/SUR2A and Kir6.2/SUR2B types of DB00171 -sensitive potassium channel . 1. We have investigated the mechanism of action of the novel anti-diabetic agent mitiglinide ( S 21403 ) on Kir6.2/ Q09428 , Kir6.2/SUR2A and Kir6.2/SUR2B types of DB00171 -sensitive potassium ( K( DB00171 ) ) channel . These possess a common pore-forming subunit , Kir6.2 , and different regulatory sulphonylurea receptor ( Q09428 ) subunits . It is believed that they correspond to native K( DB00171 ) channels in pancreatic beta-cells , heart and non-vascular smooth muscle , respectively . 2 . Kir6.2 was coexpressed with Q09428 , SUR2A or SUR2B in Xenopus oocytes and macroscopic currents were recorded in giant inside-out membrane patches . DB01252 was added to the intracellular membrane surface . 3 . DB01252 inhibited Kir6.2/ Q09428 currents at two sites : a low-affinity site on Kir6.2 and a high-affinity site on Q09428 . Low-affinity inhibition was similar for all three types of K( DB00171 ) channel but high-affinity inhibition was greater for Kir6.2/ Q09428 currents ( IC(50) , 4 nM ) than for Kir6.2/SUR2A or Kir6.2/SUR2B currents ( IC(50) , 3 and 5 microM , respectively ) . 4 . Inhibition of Kir6.2/ Q09428 currents was only slowly reversible on the time scale of electrophysiological experiments . 5 . Kir6.2/ Q09428 -S1237Y currents , which previously have been shown to lack high affinity tolbutamide inhibition , resembled Kir6.2/SUR2 currents in being unaffected by 100 nM but blocked by 10 microM mitiglinide . 6 . Our results show that mitiglinide is a high-affinity drug that shows a 1000 fold greater affinity for the beta-cell type than the cardiac and smooth muscle types of K( DB00171 ) channel , when measured in excised patches . Inhibition of human steroid 5beta-reductase ( P51857 ) by finasteride and structure of the enzyme-inhibitor complex . The Delta(4)-3-ketosteroid functionality is present in nearly all steroid hormones apart from estrogens . The first step in functionalization of the A-ring is mediated in humans by steroid 5alpha- or 5beta-reductase . DB01216 is a mechanism-based inactivator of 5alpha-reductase type 2 with subnanomolar affinity and is widely used as a therapeutic for the treatment of benign prostatic hyperplasia . It is also used for androgen deprivation in hormone-dependent prostate carcinoma , and it has been examined as a chemopreventive agent in prostate cancer . The effect of finasteride on steroid 5beta-reductase ( P51857 ) has not been previously reported . We show that finasteride competitively inhibits P51857 with low micromolar affinity but does not act as a mechanism-based inactivator . The structure of the P51857 .NADP(+)*finasteride complex determined at 1.7 A resolution shows that it is not possible for NADPH to reduce the Delta(1-2)-ene of finasteride because the cofactor and steroid are not proximal to each other . The P01024 -ketone of finasteride accepts hydrogen bonds from the catalytic residues DB00135 -58 and DB00142 -120 in the active site of P51857 , providing an explanation for the competitive inhibition observed . This is the first reported structure of finasteride bound to an enzyme involved in steroid hormone metabolism . Evidence for protein tyrosine phosphatase , tyrosine kinase , and G-protein regulation of the parallel fiber metabotropic slow EPSC of rat cerebellar Purkinje neurons . The slow EPSP ( sEPSP ) or slow EPSC ( sEPSC ) at parallel fiber to Purkinje neuron synapses is attributable to a nonselective cation channel coupled to activation of metabotropic type 1 glutamate receptors ( mGluR1s ) . Photorelease of L-glutamate in 1 msec from 4-methoxy-7-nitroindolinyl-or 7-nitroindolinyl-caged glutamate in cerebellar slices was used to isolate and study postsynaptic mechanisms coupling Q13255 to the cation channel . DB00142 immediately activated a glutamate transporter current , followed by the slow Q13255 -activated conductance . Inhibitors of kinases , phosphatases , and G-proteins were tested on the peak glutamate-evoked currents . No effects of the inhibitors were seen on the initial glutamate transporter currents . In contrast , the later Q13255 currents were either unaffected or enhanced by the protein tyrosine kinase ( PTK ) inhibitors P50391 , K252a , and staurosporine were diminished or blocked by phosphatase inhibitors but were unaffected by inhibitors of serine-threonine kinases PKA , PKC , or PKG . The selective src-PTK inhibitor P50391 ( 10 microm intracellularly ) potentiated submaximal Q13255 currents evoked by low L-glutamate concentrations but had no effect on maximal responses ( 80 or 160 microm L-glutamate ) . DB00142 -evoked Q13255 currents and parallel fiber sEPSCs were reversibly and completely inhibited by protein tyrosine phosphatase ( PTP ) inhibitor bpV(phen) ( 50-200 microm ) and by nonselective phosphatase inhibitor orthovanadate ( 0.5 or 1 mm ) . Q13255 currents were completely inhibited by GDPbetaS applied intracellularly ( 5 mm ) . The results confirm a role for a GTPase postsynaptically , show that tyrosine phosphorylation inhibits Q13255 coupling to the channel , and show that PTPs increase activation by tyrosine dephosphorylation most likely upstream of the sEPSP cation channel . UMD ( Universal mutation database ) : a generic software to build and analyze locus-specific databases . The human genome is thought to contain about 80,000 genes and presently only 3,000 are known to be implicated in genetic diseases . In the near future , the entire sequence of the human genome will be available and the development of new methods for point mutation detection will lead to a huge increase in the identification of genes and their mutations associated with genetic diseases as well as cancers , which is growing in frequency in industrial states . The collection of these mutations will be critical for researchers and clinicians to establish genotype/phenotype correlations . Other fields such as molecular epidemiology will also be developed using these new data . Consequently , the future lies not in simple repositories of locus-specific mutations but in dynamic databases linked to various computerized tools for their analysis and that can be directly queried on-line . To meet this goal , we devised a generic software called UMD ( Universal Mutation Database ) . It was developed as a generic software to create locus-specific databases ( LSDBs ) with the 4(th) Dimension(R) package from ACI . This software includes an optimized structure to assist and secure data entry and to allow the input of various clinical data . Thanks to the flexible structure of the UMD software , it has been successfully adapted to nine genes either involved in cancer ( P25054 , P04637 , P06400 , O00255 , Q09428 , P40337 , and P19544 ) or in genetic diseases ( P35555 and P01130 ) . Four new LSDBs are under construction ( P49748 , P11310 , KIR6 , and P29400 ) . Finally , the data can be transferred to core databases .