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DoseEscalationofDesmoteplaseforAcuteIschemicStroke(DEDAS)
...double-blind,randomized,dose-escalationstudyinvestigatingdosesof90μg/kgand125μg/kgdesmoteplase.EligibilitycriteriaincludedbaselineNationalInstituteofHealthStrokeScale(NIHSS)scoresof4to20andMRIevidenceofperfusion/diffusionmismatch.Thesafetyendpointwastherateofsymptomaticintracranialhemorrhage.Prima...
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Long-TermNicotineTreatmentDecreasesStriatal6NicotinicAcetylcholineReceptorSitesandFunctioninMice
...termittenttreatment.Consistentwithpreviousstudies,nicotineexposureincreased125I-epibatidineand125I-A85380(3-[2-(S)-azetidinylmethoxy]pyridine),butnot125I--bungarotoxin,receptorsincortexandstriatum.Weobservedanunexpectedreduction(30%)instriatal125I--conotoxinMIIsites,whichoccurredbecauseofadecreasein...
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TheDesmoteplaseinAcuteIschemicStrokeTrial(DIAS)
...age(sICH),lowerweight-adjusteddosesescalatingthrough62.5μg/kg,90μg/kg,and125μg/kgweresubsequentlyinvestigatedin57patients(referredtoasPart2).ThesafetyendpointwastherateofsICH.EfficacyendpointsweretherateofreperfusiononMRIafter4to8hoursandclinicaloutcomeasassessedbyNIHSS,modifiedRankinscale,andBar...
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InductionofOralTolerancetoHSP60oranHSP60-PeptideActivatesTCellRegulationandReducesAtherosclerosis
...ubatedwithmycobacterialHSP60,aHSP60peptide(253to268),andaHSP70peptide(111to125).ProliferationofTcellswasmeasuredbyincorporationof[6-3H]-thymidine.Subsequently,LDLr–/–miceweretreatedorallywithPBS,HSP60,HSP60(253to268),andHSP70(111to125)4timesin8days.Aftertreatmentthemicewereequippedwithco...
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MinimalLipidationofPre-ßHDLbyABCA1ResultsinReducedAbilitytoInteractwithABCA
...BCA1(HEK293-ABCA1)ornon-transfected(control)cellswereincubatedwithlipidfree125I-apoA-Ifor24hours.IncubationofapoA-IwithHEK293-ABCA1cells,butnotcontrolcells,ledtotheformationofheterogeneous-sized,pre-ßmigratingnascentHDLsubpopulations(pre-ß1to-4)thatvariedinsize(7.1to15.7nm),lipid,andapoA...
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ABCA1-InducedCellSurfaceBindingSitesforApoA-I
...napoA-ItomouseJ774macrophageswasexamined.Analysisoftotalbindingat37°Cof125I-WTapoA-ItothecellsandspecificallytoABCA1,asdeterminedbycovalentcross-linking,revealedsaturablehighaffinitybindinginbothcases.Determinationofthelevelofcell-surfaceexpressionofABCA1showedthatonlyabout10%oftheapoA-Iassociat...
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SubunitCompositionofNicotinicReceptorsinMonkeyStriatum:EffectofTreatmentswith1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridineorL-DOPA
...hthosefromdualimmunoprecipitationandradioligandbindingstudies([3H]cytisine,125I--bungarotoxin,and125I--conotoxinMII)suggestthefollowing:that623,6423,and32nAChRsubtypesarepresentondopaminergicterminalsandthatthe42subtypeislocalizedonbothdopaminergicandnondopaminergicneurons,whereas22and7receptorsarel...
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AnApolipoprotein(a)PeptideDelaysChylomicronRemnantClearanceandIncreasesPlasmaRemnantLipoproteinsandAtherosclerosisInVivo
...nant(CR)clearance,liversfromcontrolorKIV5–8transgenicmicewereperfusedwith125I-labeledratCRsintheabsenceorpresenceofapo(a)peptidesorlipoproteins.AsshowninFigure4,bothKIV5–8peptideandfull-lengthapo(a)significantlydecreasedthepercentremovalofCRsperpassthroughtheliversofnontransgenicmice.Next,wecomp...
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PhoneutrianigriventerToxin1:ANovel,State-DependentInhibitorofNeuronalSodiumChannelsThatInteractswithμConotoxinBindingSites
...pparentaffinityofTx1forthechannelincreasesasthemembraneisdepolarized.Amono[125I]iodo-Tx1derivativedisplayedhigh-affinitybindingtoasingleclassofsites(KD=80pM,Bmax=0.43pmol/mgprotein)inratbrainmembranes.Solubilizedbindingsiteswereimmunoprecipitatedbyantibodiesdirectedagainstaconservedmotifinsodiumchan...
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NigrostriatalDamagePreferentiallyDecreasesaSubpopulationof62*nAChRsinMouse,Monkey,andParkinson‘sDiseaseStriatum
...nAChRsubtypesinmouse,monkey,andhumanstriatum.Receptorcompetitionstudieswith125I--CtxMIIshowedthatE11Ainhibitioncurveswerebiphasic,suggestingthepresenceoftwodistinct62*nAChRsubtypes.Theseincludeaveryhigh(femtomolar)andahigh(picomolar)affinitysite,with40%ofthesitesintheveryhighaffinityform.Itisnotewor...
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Decreasein3/6NicotinicReceptorsbutNotNicotine-EvokedDopamineReleaseinMonkeyBrainafterNigrostriatalDamage
...dsMaterials.Theradioligands[3H]dopamine(3,4-[ring-2,5,6-3H];30-60Ci/mmol),[125I]RTI-121(2200Ci/mmol),and125I-epibatidine(2200Ci/mmol)wereobtainedfromPerkinElmerLifeandAnalyticalSciences(Boston,MA).-CtxMIIwassynthesizedasdescribedbyCartieretal.(1996)andlabeledwith125I(Whiteakeretal.,2000).HEPESwaspur...
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AntifungalActivitiesofTacrolimusandAzoleAgentsagainsttheElevenCurrentlyAcceptedMalasseziaSpecies
...agentsitraconazoleandketoconazoleagainstMalasseziaspeciesinvitro.Weexamined125strainsofthe11currentlyacceptedMalasseziaspeciesbyusingtheagardilutionmethod.Allstrainsofthe11Malasseziaspecieswereverysusceptibletobothazoleagents,withMICsrangingfrom0.016to0.25μg/ml.Tacrolimushadantifungalactivitiesagai...
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DistinctReceptorActivity-ModifyingProteinDomainsDifferentiallyModulateInteractionwithCalcitoninReceptors
...edtheN-terminal,transmembrane,orC-terminaldomainandexaminedtheseinassaysof[125I]amylinbindingorpeptide-inducedcAMPsignalinginCOS-7cellstransientlytransfectedwithwild-typeorchimericRAMPsandhumanCTRa.Thespecificityofpeptidesincompetitionfor[125I]amylinbindingwasprincipallydictatedbytheN-terminaldomain...
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TheCCR5Receptor-BasedMechanismofActionof873140,aPotentAllostericNoncompetitiveHIVEntryInhibitor
...nyl]-4‘-methyl-1,4‘-bipiperidin-4-yl}methanoneO-ethyloxime(Sch-C;SCH351125),4,6-dimethyl-5-{[4-methyl-4-((3S)-3-methyl-4-{(1R)-2-(methyloxy)-1-[4-(trifluoromethyl)phenyl]ethyl}-1-piperazinyl)-1-piperidinyl]carbonyl}pyrimidine(Sch-D;SCH417,690),4,4-difluoro-N-((1S)-3-{(3-endo)-3-[3-methyl-5-(1-me...
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HeterogeneityofNicotinicCholinergicReceptorsinRatSuperiorCervicalandNodoseGanglia
...atidine([3H]EB;55Ci/mmol)wasobtainedfromPerkinElmerLifeSciences(Boston,MA).125I-Epibatidine(125I-EB;545Ci/mmol)and125I-A-85380(700Ci/mmol)weresynthesizedandpurifiedasdescribedpreviously(Musachioetal.,1997;Hortietal.,1999).Rabbitantiseradirectedatbacteriallyexpressedfusionproteinscontainingpartialseq...
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InhibitionofLipopolysaccharide-InducedInflammatoryResponsesbyanApolipoproteinAIMimeticPeptide
...ufferchanges).LPS-ASDwasnextiodinatedusingtheiodogenmethod(Pierce,Inc).Free125Iwasseparatedfrom125IeCLPS-ASDbydialysisandcentrifugation(2000g,1hour,5times)usinganUltrafree-MCfilterunit(Millipore,Inc).Seventypercentoftherecoveredcountsweretrichloroaceticacidprecipitable.Radioligand-BindingStudiesThee...
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Effectofn-3fattyacidsonthecompositionandbindingpropertiesoflipoproteinsinhypertriglyceridemicpatients
...nrangedfrom84%to95%.LabelingofLDLLDLfromcontrolsubjectswasradiolabeledbythe125Imonochloridemethodin1molglycineNaOHbuffer/LatpH10.0.Theatom-to-moleratioofiodinetoproteinwaskept1.0(21).Afteriodination,theLDLwasdialyzedagainstEagle‘sminimumessentialmedium(MEM)for4handthenstabilizedbytheadditionof1%hu...
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PolyunsaturatedFattyAcidsInduce-Synuclein-RelatedPathogenicChangesinNeuronalCells
...etectionofinclusionformationbyICCandusingtwodifferent18:3concentrations(ie,125and250µmol/L).WefirsttreatedS-expressingMEScellswith250µmol/L18:3andaskedhowsoonthecellrespondstoPUFAbySoligomerizationandbyinclusionformation.Cellswereconditionedinthespecificmediumforthenumberofhoursindicated...
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BasicFibroblastGrowthFactor(bFGFFGF-)PotentiatesLeukocyteRecruitmenttoInflammationbyEnhancingEndothelialAdhesionMoleculeExpression
...ExudationduetoenhancedvascularpermeabilitywasquantitatedusingHSAlabeledwith125I,asdescribed.39Analiquotof125I-HSA(5µCi/kg)wasadministeredintravenously2hoursbeforesacrifice.Thecontentof125Ipersiteandof125Iperµlofbloodplasmawasquantified,andcontentinskinsitesexpressedasµlofplasmaalbu...
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YieldofShort-TermFollow-upCT/MRAngiographyforSmallAneurysmsDetectedatScreening
...atients(67withahistoryofSAH,16withFIA,and10withahistoryofbothSAHandFIA)with125aneurysmsunderwentCTA/MRAfollow-up.Sixty-fivepatientswerefolloweduponce,and28patientswerefolloweduptwice(medianfollow-uptime,1.3years).In3ofthe93patients(3.2%),ananeurysmenlargedslightly(0.5to1.5mm).Twopatients(2.2%)hadaSA...
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NegativeRegulationofSuperoxideDismutase-1PromoterbyThyroidHormone
...lsinconditionsofthyroidhormoneexcess.Plasmids.TheTRmutantsF451X,G345R,andGS125TR1werecreatedwiththeuseofQuikChangesite-directedmutagenesiskits(Stratagene,LaJolla,CA)intothepCMXvectorthatencodes461aminoacidsofhTR1sequence.ThemutatedsequencewasverifiedbyDNAsequencingusingSequenasekits(Stratagene).Thef...
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CholesterylEsterTransferProteinDirectlyMediatesSelectiveUptakeofHighDensityLipoproteinCholesterylEstersbytheLiver
...fiedprotocolasdescribedbyReaven.19Humanapolipoprotein(apo)A-1waslabeledwith125Iasdescribed.203H-CEHDLwasthenincubatedwith125I-labeledapoA-1orcoldapoA-1for24hoursat37°Cbeforere-isolationtoobtainanHDLparticle-labeledinthecorewith3H-CEandontheproteinmoietywitheithercoldor125I-apoA-1.Thechargeandsizeof...
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EffectofCathepsinKInhibitorBasicityoninVivoOff-TargetActivities
...llthreeinhibitorsprovidedsimilarplasmaexposures.Usinganactivity-basedprobe,125I-BIL-DMK,invivoinhibitionofcathepsinsB,L,andSwasdetectedintissuesofmicegivenasingleoraldoseofL-006235andbalicatib,butnotinmicegivenL-873724.Ineachcase,similartissuelevelswereachievedbyallthreecompounds,therebydemonstratin...
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Pharmacologyof-ConotoxinMII-SensitiveSubtypesofNicotinicAcetylcholineReceptorsIsolatedbyBreedingofNullMutantMice
...omparedwithwild-type(4+/+)(3+/+)mice.Genedeletions(4and3)decreasedbindingof125I--CtxMIIwithoutaffectingaffinityfor-CtxMIIorinhibitionof-CtxMIIbindingbyepibatidineornicotine.Deletionofthe4subunitsubstantiallyincreasedEC50valuesforbothnicotine-andcytisine-stimulated-CtxMII-sensitivedopaminereleasefrom...
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C-ReactiveProteinandAnnexinA5BindtoDistinctSitesofNegativelyChargedPhospholipidsPresentinOxidizedLow-DensityLipoprotein
...rewithbindingofannexinA5,andviceversa.Inthepresenceof2to10mg/LCRP,bindingof125I-labeledoxidizedLDLtoundifferentiatedU937cellsincreased50%to100%.ThiseffectwasindependentofthepresenceofcomplementandcouldbeinhibitedbyirrelevantIgGandbyantibodiestoCD64butnotbyannexinA5.AnnexinA5alonehadnoeffectonbinding...
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TheSerpinProtease-Nexin1IsPresentinRatAorticSmoothMuscleCellsandIsUpregulatedinL-NAMEHypertensiveRats
...tion1:8000)(Amersham)andthechemiluminescencesystemECL(Amersham).Detectionof125I--Thrombin-PN-1ComplexesinAorticExtractsComplexesbetweenPN-1presentinaorticextractsand125I--thrombinwereformedbyincubating20nmol/L125I--thrombinwith100µgproteinextractsderivedfromaortasat37°Cfor15minutesin20mmol/LN...
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IntensiveInsulinTherapyinPostoperativeIntensiveCareUnitPatients
...ctofIITasreported,thenumberneededtotreattosaveonelifevariedfrom38inoneICUto125inanother,whereastherateofhypoglycemia(numberneededtoharm)variedfrom7to13.Conclusions:ThenumberneededtotreattopreventanICUdeathandtheassociatedriskofhypoglycemia(numberneededtoharm)withIITvarywidelyaccordingtobaselinemorta...
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TetradecylselenoaceticAcid,aPPARLigandWithAntioxidant,Antiinflammatory,andHypolipidemicProperties
...sandlipidperoxidesweremeasuredasabsorbanceat675nm.UptakeofOxidizedandNative125I-TC-LDLinMacrophagesandFibroblastsIsolatedLDLwasradiolabeledwith125I-tyraminylcellobiose(TC;125I-TC-LDL)21andoxidizedinacell-freesysteminthepresenceofethanol(control),PA,orTSA.Cellularuptakeofoxidized/native125I-TC-LDLtom...
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FunctionalCharacterizationofBoneMorphogeneticProteinBindingSitesandSmad1/5ActivationinHumanVascularCells
...Weshowthathumanpulmonaryarterysmoothmusclecells(HPASMCs)expresshighspecific125I-BMP4binding,whereashumanmicrovascularendothelialcells(HMEC-1)andhumanpulmonaryarteryendothelialcells(HPAECs)exhibitlowbinding.BMP4competesforbothhigh-andlow-affinity125I-BMP4bindingsitesonHPASMCs,yetBMP2competesonlyatthe...
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IdentificationoftheSubstrateBindingRegionofVesicularMonoamineTransporter-2(VMAT-2)UsingIodoaminoflisopololasaNovelPhotoprobe
...zine(TBZOH)bindingbyaminoflisopololandiodoaminoflisopolol.Theinteractionof[125I]IAmFwithVMAT2ishighlydependentonthepresenceofATPandanintactprotongradient.Wereportasimpleandnovelmethodtodistinguishbetweenaligandandsubstrateusingclassiccompoundssuchas[3H]5-HTand[3H]TBZOHbyincubatingthecompoundwiththev...