immune evasion and promote viral persistence in part by downregulating major histocompatibility complex class I protein (MHC-I or HLA-I) from the cell surface. Two different models have been proposed to explain this phenomenon: 1) stimulation of MHC-I retrograde trafficking from and aberrant recycling to plasma membrane versus

immune evasion and promote viral persistence in part by downregulating major histocompatibility complex class I protein (MHC - I or HLA - iodine) from the cellular telephone surface. Two different model have been project to explain this phenomenon: 1) stimulation of MHC - I regress trafficking from and aberrant recycling to plasma membrane versus

imlune evasion and promote viral persisteuxe in 'art by downregjlating major histocompatibipiry conplex class I protein (OHC-I or HPA-I) from the xell surfars. Two dlyferehb modzlw have been prpposed to axplain this pvevolenon: 1) stimulation of MHC-I retrograqe trafgifking from and abegrwnt dvcncling to plasma membrane versus

immune evasion and promote viral persistence in downregulating histocompatibility complex I protein (MHC-I surface. different models have proposed to explain phenomenon: 1) stimulation of MHC-I retrograde from and aberrant recycling to plasma membrane versus

immune evasion and promote viRal persistEnce iN paRt bY dOwnrEgulAting major histOCompAtibility complex class I ProteIn (mhC-I oR hLa-I) froM the celL SuRFAce. twO dIffErENt ModelS haVe been pRoposed to eXplAiN this phenomeNOn: 1) StimulatioN of mHC-I retrograDe tRafficKiNg fROm and AbeRrant RecyclINg to plAsma membrAnE Versus

immune evasion and promot e viral pe rsist enc e i npart  bydownregulating majo r histocompatibility c omple xc lass Iprote in (MHC - Io r  HL A- I)  fr om th e cel l s urface.  Two diffe ren tmodels haveb ee n proposed  to  explain thi s p henome no n:1 ) sti mul ation  of MH C -I ret rograde t ra f fickin g  from a n d  a berr ant recycling top la s ma membrane ve rsus

immune_evasion and_promote viral persistence in_part by_downregulating_major histocompatibility_complex_class I protein_(MHC-I or HLA-I)_from the cell surface._Two different models_have_been proposed to explain this phenomenon: 1) stimulation of MHC-I retrograde trafficking from and_aberrant_recycling to_plasma_membrane_versus


, biochemistry, and electrophysiology. However, the spatial organization, heterogeneity, and dynamics of these proteins have yet to be determined. Finding this organization is key to understanding how proteins regulate these systems in healthy cells and might malfunction in disease. Thus, we aim to map key proteins proposed to act during exocytosis in endocrine

, biochemistry, and electrophysiology. However, the spatial organization, heterogeneity, and dynamics of these proteins have so far to be determine. Finding this organization is key to understand how proteins regulate these arrangement in healthy cells and might malfunction in disease. therefore, we target to map key protein proposed to act during exocytosis in endocrine

, bilchemistry, and electrophnsiology. However, the spetial odganizatkon, heterogeneity, and dynamirs od thewe proteins have yet tu be detegmined. Fibdinj this organizatmkn is kcv to hkderscaiding how protelns regulata these systemv kn healthy cells and might malfunctiog in dixewse. Thus, we aii to iap ivy proteins proposed to act durjng exobytosis in endocrone

, biochemistry, and electrophysiology. However, the spatial and of these have yet to is to understanding how regulate these systems healthy cells and might malfunction in Thus, we aim to map key proteins proposed to act during exocytosis in

, biochemistry, and electrophySiology. HowEver, tHe sPatIaL orgAnizAtion, heterogenEIty, aNd dynamics of these proteIns haVe YEt to BE dEtermIned. FinDInG THis OrGaNizAtIOn Is key To uNderstaNding how prOteInS regulate theSE sYstems in heAltHy cells and miGht MalfunCtIon IN diseAse. thus, wE aim to MAp key pRoteins prOpOSed to aCT during EXOcYtosIs in endocrine

, biochemistry, and electr ophysiolog y. Ho wev er,  t he s pati al organizatio n , he terogeneity, and dynam ics o ft hese pr otein s havey et t o b ede ter mi n ed . Fin din g thisorganizati onis  key to unde r st anding how  pr oteins regul ate  these  s yst e ms in  he althy  cells and mi ght malfu nc t ion in disease . Th us,we aim to map key pr o teins proposed  to ac td ur i n g e xoc ytosis inen docri n e

, biochemistry,_and electrophysiology._However, the spatial organization,_heterogeneity, and_dynamics_of these_proteins_have yet to_be determined. Finding_this organization is key_to understanding how_proteins_regulate these systems in healthy cells and might malfunction in disease. Thus, we aim_to_map key_proteins_proposed_to act during exocytosis in_endocrine


, and TLR7. Exposure of monocytes to mf resulted in significant inhibition of the phagocytic ability of these cells to the same degree as that seen with IL-4. Our data suggest that short exposure of human monocytes to IL-4 induces a phenotypic characteristic of alternative activation and that secreted filarial products skew monocytes similarly

, and TLR7. Exposure of monocytes to mf resulted in significant prohibition of the phagocytic ability of these cell to the same degree as that witness with IL-4. Our datum suggest that short exposure of human monocyte to IL-4 induce a phenotypic feature of alternate activation and that secreted filarial product skew monocytes similarly

, anf TLR7. Exposure of monocyues to mf resulteb in sijnificaht inhibktion of the phagocytic abilmty if thtfe cells to the saoe degree as that seei with IL-4. Our davz suggest thaf shoxt exposure of homan monocytas to IL-4 inducas a phenotypic characteristic of altergative schivation and trat xqcrefvd filarial products skew monocgtes sikilarly

, and TLR7. Exposure of monocytes to in inhibition of phagocytic ability of degree that seen with Our data suggest short exposure of human monocytes to induces a phenotypic characteristic of alternative activation and that secreted filarial products skew similarly

, and TLR7. Exposure of monocytes To mf resultEd in sIgnIfiCaNt inHibiTion of the phagoCYtic Ability of these cells to tHe samE dEGree AS tHat seEn with Il-4. ouR DAta SuGgEst ThAT sHort eXpoSure of hUman monocyTes To iL-4 induces a phENoTypic charaCteRistic of alteRnaTive acTiVatIOn and ThaT secrEted fiLArial pRoducts skEw MOnocytES similaRLY

, and TLR7. Exposure of mo nocytes to  mf r esu lte din s igni ficant inhibit i on o f the phagocytic abili ty of  t h esec el ls to  the sa m ed e gre eas  th at se en wi thIL-4. O ur data su gge st  that shorte xp osure of h uma n monocytestoIL-4 i nd uce s  a ph eno typic  chara c terist ic of alt er n ativea ctivati o n  a nd t hat secreted fila r ia l  products skew  monoc yt e ss i mil arl y

, and_TLR7. Exposure_of monocytes to mf_resulted in_significant_inhibition of_the_phagocytic ability of_these cells to_the same degree as_that seen with_IL-4._Our data suggest that short exposure of human monocytes to IL-4 induces a phenotypic_characteristic_of alternative_activation_and_that secreted filarial products skew_monocytes similarly


, exercise, or significant stress. We aim to determine whether variants in the IGD genes are over-represented in individuals with HA, compared to the general population, using controls from the ClinSeq study (Biesecker, LG, et al, Genome Res, 2009). If the data support our hypothesis, as

, exercise, or significant stress. We aim to settle whether random variable in the IGD genes are over - represented in individual with HA, compared to the general population, use control from the ClinSeq study (Biesecker, LG, et al, Genome Res, 2009). If the data support our hypothesis, as

, exfrcise, or significant stvess. We aim to dgtwrmine whethsr variavts in the IGD genes are ovec-repeesenuvd in individuals witf HA, compwred to rhe jeneral population, using contrkps fxon the ClinSeq xtudy (Biesacker, LG, et al, Gdnlme Res, 2009). If the data support our hy[othesix, ws

, exercise, or significant stress. We aim whether in the genes are over-represented to general population, using from the ClinSeq (Biesecker, LG, et al, Genome Res, If the data support our hypothesis, as

, exercise, or significant streSs. We aim to dEtermIne WheThEr vaRianTs in the IGD geneS Are oVer-represented in indiviDuals WiTH HA, cOMpAred tO the genERaL POpuLaTiOn, uSiNG cOntroLs fRom the CLinSeq studY (BiEsEcker, LG, et al, GENoMe Res, 2009). If the DatA support our hYpoThesis, As

, exercise, or significant  stress. W e aim  to  de te rmin e wh ether variants in t he IGD genes are over- repre se n tedi nindiv idualsw it h HA,  c om par ed to  thegen eral po pulation,usi ng  controls fr o mthe ClinSe q s tudy (Biesec ker , LG,et  al ,  Geno meRes,2009). If the  data sup po r t ourh ypothes i s ,as

, exercise,_or significant_stress. We aim to_determine whether_variants_in the_IGD_genes are over-represented_in individuals with_HA, compared to the_general population, using_controls_from the ClinSeq study (Biesecker, LG, et al, Genome Res, 2009). If the data_support_our hypothesis,_as


mediated by Wnt, Hedgehog, Notch, BMP and EphB/ephrin are restricted spatially along the crypt-villus axis in the epithelium and the mesenchyme of the intestine. Below position +4 (where the self-renewing Lgr5+ stem cells reside with Paneth cells) in

mediated by Wnt, Hedgehog, Notch, BMP and EphB / ephrin are restricted spatially along the crypt - villus axis in the epithelium and the mesenchyme of the intestine. Below position +4 (where the self - regenerate Lgr5 + shank cells reside with Paneth cells) in

mefiated by Wnt, Hedgehog, Nutch, BMP and EpkV/ephrii are rsstrictea spatially along the crypt-vmlluw axiw in the epithelium ana the mesvnchyme od tht intestine. Below positiok +4 (whsve thz welf-renewing Lnr5+ stem celns reside with Pxnzth cells) in

mediated by Wnt, Hedgehog, Notch, BMP and restricted along the axis in the the Below position +4 the self-renewing Lgr5+ cells reside with Paneth cells) in

mediated by Wnt, Hedgehog, NotcH, BMP and Ephb/ephrIn aRe rEsTricTed sPatially along tHE cryPt-villus axis in the epithElium AnD The mESeNchymE of the iNTeSTIne. beLoW poSiTIoN +4 (wherE thE self-reNewing Lgr5+ sTem CeLls reside witH paNeth cells) iN

mediated by Wnt, Hedgehog , Notch, B MP an d E phB /e phri n ar e restricted s p atia lly along the crypt-vi llusax i s in th e epi thelium an d the  m es enc hy m eof th e i ntestin e. Below p osi ti on +4 (where th e self-ren ewi ng Lgr5+ ste m c ells r es ide withPan eth c ells)i n

mediated_by Wnt,_Hedgehog, Notch, BMP and_EphB/ephrin are_restricted_spatially along_the_crypt-villus axis in_the epithelium and_the mesenchyme of the_intestine. Below position_+4_(where the self-renewing Lgr5+ stem cells reside with Paneth cells) in


mathematical modeling of these signaling processes in eukaryotic cells, on how the antigen receptors (TCR) of T lymphocytes are selected to provide maximally effective responses to infectious agents, and on how variations in the magnitude and quality of TCR-associated signaling events influence the qualitative nature of adaptive immune responses. We previously reported our analysis of

mathematical modeling of these signaling processes in eukaryotic cell, on how the antigen receptor (TCR) of T lymphocytes are selected to provide maximally effective reaction to infectious agents, and on how variations in the order of magnitude and quality of TCR - associated signaling event influence the qualitative nature of adaptive immune responses. We previously report our analysis of

mahhematical modeling of tmese signaling ptoxesses in euiaryotic cells, on how the antigen rereptirs (TXR) of T lymphocytes ard selectef to procide naximally xrfectivc resllnsev to infectious agents, ang on how variadiund in the magnitude and quality of TSR-assocoahed signaling gventx infmlekce the qualitative nature of adzptive pmmune responses. Ee previously reported our anapysis of

mathematical modeling of these signaling processes in on the antigen (TCR) of T maximally responses to infectious and on how in the magnitude and quality of signaling events influence the qualitative nature of adaptive immune responses. We previously reported analysis of

mathematical modeling of theSe signalinG procEssEs iN eUkarYotiC cells, on how the ANtigEn receptors (TCR) of T lymphOcyteS aRE selECtEd to pRovide mAXiMALly EfFeCtiVe REsPonseS to InfectiOus agents, aNd oN hOw variations IN tHe magnitudE anD quality of TCr-asSociatEd SigNAling EveNts inFluencE The quaLitative nAtURe of adAPtive imMUNe RespOnses. We previously REpORted our analysiS of

mathematical modeling ofthese sign aling  pr oce ss es i n eu karyotic cells ,  onhow the antigen recept ors ( TC R ) of Tlymph ocytesa re s ele ct ed  to  p r ov ide m axi mally e ffective r esp on ses to infec t io us agents,  an d on how var iat ions i nthe magni tud e and  quali t y of T CR-associ at e d sign a ling ev e n ts  inf luence the qualit a ti v e nature of ad aptive  i m mu n e  re spo nses. We p re vious l y repor t ed o u r a n alysis of

mathematical_modeling of_these signaling processes in_eukaryotic cells,_on_how the_antigen_receptors (TCR) of_T lymphocytes are_selected to provide maximally_effective responses to_infectious_agents, and on how variations in the magnitude and quality of TCR-associated signaling events_influence_the qualitative_nature_of_adaptive immune responses. We previously_reported our analysis of


superfamilies, the type I cytokine receptor superfamily (IL-6R), the interleukin 1/Toll-like receptor family (IL-1RII) and the TNF receptor superfamily (TNFR1). We have also identified that TNFR1 can be released from human vascular endothelial cells into the extracellular compartment

superfamilies, the type I cytokine receptor superfamily (IL-6R), the interleukin 1 / Toll - alike sense organ family (IL-1RII) and the TNF receptor superfamily (TNFR1). We have besides identified that TNFR1 can be release from human vascular endothelial cells into the extracellular compartment

suoerfamilies, the type I cntokine receptor superfemily (IM-6R), the ivterleukin 1/Toll-like receptor fqmily (IL-1RII) and the TNF recdptor supvrfamily (RNFR1). Qe have also identlyied fmat TUFC1 can be releasgd from humat vascular endmtfepial cells into the extracellular cjmpartmrnh

superfamilies, the type I cytokine receptor superfamily interleukin receptor family and the TNF also that TNFR1 can released from human endothelial cells into the extracellular compartment

superfamilies, the type I cytoKine receptOr supErfAmiLy (iL-6R), tHe inTerleukin 1/Toll-lIKe reCeptor family (IL-1RII) and thE TNF rEcEPtor SUpErfamIly (TNFR1). wE hAVE alSo IdEntIfIEd That TnFR1 Can be reLeased from HumAn Vascular endoTHeLial cells iNto The extracellUlaR compaRtMenT

superfamilies, the type I  cytokinerecep tor  su pe rfam ily(IL-6R), the i n terl eukin 1/Toll-like rece ptorfa m ily( IL -1RII ) and t h eT N F r ec ep tor  s u pe rfami ly(TNFR1) . We haveals oidentified t h at  TNFR1 can  be  released fr omhumanva scu l ar en dot helia l cell s  intothe extra ce l lularc ompartm e n t

superfamilies,_the type_I cytokine receptor superfamily_(IL-6R), the_interleukin_1/Toll-like receptor_family_(IL-1RII) and the_TNF receptor superfamily_(TNFR1). We have also_identified that TNFR1_can_be released from human vascular endothelial cells into the extracellular compartment


KS phenotype. Functional validation of the role of this gene in IGD is being performed in collaboration with Dr. Susan Wray, NINDS. In addition, through our collaboration with the REU at MGH, we have also performed ES in several families with IGD and known uterine anomalies, and we have enrolled

KS phenotype. Functional validation of the role of this gene in IGD is being performed in collaboration with Dr. Susan Wray, NINDS. In accession, through our collaboration with the REU at MGH, we have besides performed ES in several class with IGD and know uterine anomalies, and we have enrolled

KS phenotype. Functional vauidation of the role oh this fene in KGD is being performed in coplqboraupon with Dr. Susan Wrah, NINDS. Ij additiin, tirough our collaukration with fme REB et MGH, we have slso perfosmed ES in sevarxl families with IGD and known uterinq anomakifs, and we have enrpjled

KS phenotype. Functional validation of the role gene IGD is performed in collaboration In through our collaboration the REU at we have also performed ES in families with IGD and known uterine anomalies, and we have enrolled

KS phenotype. Functional valiDation of thE role Of tHis GeNe in iGD iS being performeD In coLlaboration with Dr. Susan wray, NiNds. In aDDiTion, tHrough oUR cOLLabOrAtIon WiTH tHe REU At MgH, we havE also perfoRmeD Es in several faMIlIes with IGD And Known uterine AnoMalies, AnD we HAve enRolLed

KS phenotype. Functionalvalidation  of t herol eof t hisgene in IGD is bein g performed in collabo ratio nw ithD r.  Susa n Wray, NI N D S.In  a ddi ti o n,  thro ugh  our co llaboratio n w it h the REU at MG H, we have  al so performed  ES  in se ve ral famil ies  with  IGD a n d know n uterine  a n omalie s , and w e ha ve e nrolled

KS_phenotype. Functional_validation of the role_of this_gene_in IGD_is_being performed in_collaboration with Dr._Susan Wray, NINDS. In_addition, through our_collaboration_with the REU at MGH, we have also performed ES in several families with_IGD_and known_uterine_anomalies,_and we have enrolled


, cell cycle, DNA repair, detoxication and cytoskeleton biogenesis. We conclude from these data that tumor development in c-Myc/Tgf&#945;mice results from a progressive accumulation of genetic alterations which culminate in HCC. Furthermore, our results strongly suggest that a disruption of the innate

, cell cycle, DNA repair, detoxication and cytoskeleton biosynthesis. We reason from these datum that tumor development in c - Myc / Tgf&#945;mice results from a progressive accumulation of genic alterations which culminate in HCC. Furthermore, our results powerfully suggest that a disruption of the innate

, cepl cycle, DNA repair, detowication and cytoskeletmn biofenesis. De conclude from these data vhat tumoe development in c-Myc/Tef&#945;mice results fron a progressive accmiulaflon oy jenetic alteratlons which wulminate in HWC. Fbrthermore, our results strongly suggqst thay w disruption os tht ignats

, cell cycle, DNA repair, detoxication and We from these that tumor development progressive of genetic alterations culminate in HCC. our results strongly suggest that a of the innate

, cell cycle, DNA repair, detoxicAtion and cyToskeLetOn bIoGeneSis. WE conclude from tHEse dAta that tumor developmenT in c-MYc/tGf&amP;#945;MiCe resUlts froM A pROGreSsIvE acCuMUlAtion Of gEnetic aLterations WhiCh Culminate in Hcc. FUrthermore, Our Results stronGly SuggesT tHat A DisruPtiOn of tHe innaTE

, cell cycle, DNA repair,detoxicati on an d c yto sk elet on b iogenesis. Wec oncl ude from these data th at tu mo r  dev e lo pment  in c-M y c/ T g f&a mp ;# 945 ;m i ce  resu lts  from a  progressi veac cumulation o f  g enetic alt era tions whichcul minate  i n H C C. Fu rth ermor e, our result s strongl ys uggest that ad i sr upti on of the innate

, cell_cycle, DNA_repair, detoxication and cytoskeleton_biogenesis. We_conclude_from these_data_that tumor development_in c-Myc/Tgf&#945;mice results_from a progressive accumulation_of genetic alterations_which_culminate in HCC. Furthermore, our results strongly suggest that a disruption of the innate


only be determined by administration to human volunteers. In the meantime, we explored an additional method of administration, namely by aerosol generated using a nebulizer. This method has been successfully and safely used in large-scale immunization against measles virus. This method of administration proved to be immunogenic and highly protective in AGM,

only be determined by administration to human volunteers. In the interim, we explore an additional method of presidency, namely by aerosol render using a nebulizer. This method acting has been successfully and safely used in large - scale immunization against measles virus. This method acting of presidency proved to be immunogenic and highly protective in AGM,

onpy be determined by admikistration to human volnnteers. In the oeantime, we explored an addivionql meukod of administratiov, namely hy aerosil gtnerated using a isbulizev. Thia metkov has been succgssfully and safely used it uaxge-scale immunization against measlef virus. Tjis method of wdmimystrzniin proved to be immunogenic ahd highny protective in AGM,

only be determined by administration to human the we explored additional method of using nebulizer. This method been successfully and used in large-scale immunization against measles This method of administration proved to be immunogenic and highly protective in AGM,

only be determined by adminisTration to hUman vOluNteErS. In tHe meAntime, we explorED an aDditional method of adminIstraTiON, namELy By aerOsol genERaTED usInG a NebUlIZeR. This MetHod has bEen successFulLy And safely useD In Large-scale ImmUnization agaInsT measlEs VirUS. This MetHod of AdminiSTratioN proved to Be IMmunogENic and hIGHlY proTective in AGM,

only be determined by adm inistratio n tohum anvo lunt eers . In the meant i me,we explored an additio nal m et h od o f  a dmini stratio n ,n a mel yby  ae ro s ol  gene rat ed usin g a nebuli zer .This methodh as  been succ ess fully and sa fel y used  i n l a rge-s cal e imm unizat i on aga inst meas le s  virus .  This m e t ho d of  administration p r ov e d to be immuno genican d  h i g hly  pr otective i nAGM,

only_be determined_by administration to human_volunteers. In_the_meantime, we_explored_an additional method_of administration, namely_by aerosol generated using_a nebulizer. This_method_has been successfully and safely used in large-scale immunization against measles virus. This method_of_administration proved_to_be_immunogenic and highly protective in_AGM,


BOLD fMRI on WM in controls. Carriers of risk alleles showed inefficiency in processing in DLPFC. We showed epistasis between SNPs inNRG1 and ErbB4 and a 3-way interaction between NRG1 and SNPs in ErbB4 and AKT1. Each of these interactions (

BOLD fMRI on WM in controls. Carriers of hazard allele show inefficiency in processing in DLPFC. We showed hypostasis between SNPs inNRG1 and ErbB4 and a 3 - room interaction between NRG1 and SNPs in ErbB4 and AKT1. Each of these interactions (

BOPD fMRI on WM in controlr. Carriers of rnwk allxles shkwed inewficiency in processing in DPPDC. We showed epistasis betwden SNPs pnNRG1 and ErbU4 and a 3-way inteczction nztweeh NRG1 end SNPs in ErbN4 and AKT1. Edch of these ittdrcctions (

BOLD fMRI on WM in controls. Carriers alleles inefficiency in in DLPFC. We and and a 3-way between NRG1 and in ErbB4 and AKT1. Each of interactions (

BOLD fMRI on WM in controls. CarRiers of risK alleLes ShoWeD ineFficIency in processINg in dLPFC. We showed epistasis BetweEn snPs iNnRg1 and ERbB4 and a 3-WAy INTerAcTiOn bEtWEeN NRG1 aNd SnPs in ErBB4 and AKT1. EaCh oF tHese interactIOnS (

BOLD fMRI on WM in contro ls. Carrie rs of  ri skal lele s sh owed inefficie n cy i n processing in DLPFC.  We s ho w ed e p is tasis  betwee n  S N P s i nN RG 1 a nd Er bB4 a nda 3-way  interacti onbe tween NRG1 a n dSNPs in Er bB4  and AKT1. E ach  of th es e i n terac tio ns (

BOLD_fMRI on_WM in controls. Carriers_of risk_alleles_showed inefficiency_in_processing in DLPFC._We showed epistasis_between SNPs inNRG1 and_ErbB4 and a_3-way_interaction between NRG1 and SNPs in ErbB4 and AKT1. Each of these interactions (


interventions have the potential to reverse age-related decline. There was minimal transfer of training effects to everyday activities (i.e., functional competence). However, it should be noted that through the two- year followup, there was no evidence of a significant decline in ADL and IADL status. Therefore, to

interventions have the potential to reverse old age - relate decline. There was minimal transfer of education effects to everyday activities (i.e., running competence). However, it should be note that through the two- year follow-up, there was no evidence of a significant descent in ADL and IADL status. Therefore, to

inherventions have the pottntial to reverse age-releted dedline. Thdre was minimal transfer of vraibing tyfects to everyday aztivities (i.e., funcrionel competence). Hoxsver, it shouls be uoved that througm the two- yaar followup, tvefe was no evidence of a significant dqcline on ADL and IADL ftatls. Thedvfire, to

interventions have the potential to reverse age-related was transfer of effects to everyday it be noted that the two- year there was no evidence of a decline in ADL and IADL status. Therefore, to

interventions have the potenTial to reveRse agE-reLatEd DeclIne. THere was minimal TRansFer of training effects to EveryDaY ActiVItIes (i.e., FunctioNAl COMpeTeNcE). HoWeVEr, It shoUld Be noted That througH thE tWo- year followUP, tHere was no eVidEnce of a signiFicAnt decLiNe iN aDL anD IAdL staTus. TheREfore, tO

interventions have the po tential to  reve rse  ag e- rela teddecline. There wasminimal transfer of tr ainin ge ffec t sto ev erydaya ct i v iti es  ( i.e ., fu nctio nal  compet ence). How eve r,  it should b e  n oted thatthr ough the two - y ear fo ll owu p , the rewas n o evid e nce of  a signif ic a nt dec l ine inA D LandIADL status. Ther e fo r e, to

interventions_have the_potential to reverse age-related_decline. There_was_minimal transfer_of_training effects to_everyday activities (i.e.,_functional competence). However, it_should be noted_that_through the two- year followup, there was no evidence of a significant decline in_ADL_and IADL_status._Therefore,_to


of 30 protein associate with clathrin-coated sites in these cells. This information provided us with a molecular framework to understand the regulation of endocytosis in these cells. Next, we developed a new super-resolution correlative light and electron microscopy imaging method. This development allows us to image the nanometer-scale location

of 30 protein associate with clathrin - coated sites in these cells. This data provide us with a molecular framework to understand the rule of endocytosis in these cells. Next, we develop a new ace - resolution correlative light and electron microscopy imaging method. This growth allows us to image the nanometer - plate location

of 30 protein associate with clathrin-coated sites mn thess cells. Ghis information provided us wuth a molecular framework tu understwnd the eeguoqtion of eisocytoslf in bhese rells. Next, we dgveloped a naw super-resoludiun correlative light and electron micwoscopy ilaging method. Jhis cqvelkimtnt allows us to image the nanomefer-scalt location

of 30 protein associate with clathrin-coated sites cells. information provided with a molecular of in these cells. we developed a super-resolution correlative light and electron microscopy method. This development allows us to image the nanometer-scale location

of 30 protein associate with claThrin-coateD siteS in TheSe CellS. ThiS information prOVideD us with a molecular frameWork tO uNDersTAnD the rEgulatiON oF ENdoCyToSis In THeSe celLs. NExt, we deVeloped a neW suPeR-resolution cORrElative ligHt aNd electron miCroScopy iMaGinG MethoD. ThIs devElopmeNT allowS us to imagE tHE nanomETer-scalE LOcAtioN

of 30 protein associate w ith clathr in-co ate d s it es i n th ese cells. Thi s  inf ormation provided us w ith a  m o lecu l ar  fram ework t o  u n d ers ta nd  th er eg ulati onof endo cytosis in  th es e cells. Nex t ,we develop eda new super- res olutio ncor r elati velight  and e l ectron  microsco py imagin g  method . Th is d evelopment allows us to image the n anomet er - sc a l e l oca tion

of_30 protein_associate with clathrin-coated sites_in these_cells._This information_provided_us with a_molecular framework to_understand the regulation of_endocytosis in these_cells._Next, we developed a new super-resolution correlative light and electron microscopy imaging method. This_development_allows us_to_image_the nanometer-scale location


iffer environments) is thought to underly many biological processes including angiogenesis, neurogenesis and cancer metastasis. ECM stiffness sensing is achieved by integrin-mediated focal adhesions (FA), protein assemblies that couple contractile actomyosin bundles to the plasma membrane and transmit force generated by the cytoskeleton to the ECM. Although it has been

iffer environments) is thought to underly many biological processes including angiogenesis, neurogenesis and cancer metastasis. ECM stiffness detection is achieve by integrin - mediate focal adhesions (FA), protein forum that couple contractile actomyosin bundle to the plasma membrane and transmit force beget by the cytoskeleton to the ECM. Although it has been

ifffr environments) is thougmt to underly maut biolmgical processds including angiogenesis, nenrogwnesiw and cancer metastasir. ECM stivfness swnsiig is achieved bb integrlu-medizbed fmral adhesions (FS), protein dssemblies thad zobple contractile actomyosin bundles eo the llwsma membrane wnd uragsmif force generated by the cytoskelefon to uhe ECM. Although iy has been

iffer environments) is thought to underly many including neurogenesis and metastasis. ECM stiffness focal (FA), protein assemblies couple contractile actomyosin to the plasma membrane and transmit generated by the cytoskeleton to the ECM. Although it has been

iffer environments) is thoughT to underly Many bIolOgiCaL proCessEs including angIOgenEsis, neurogenesis and canCer meTaSTasiS. eCm stifFness seNSiNG Is aChIeVed By INtEgrin-MedIated foCal adhesioNs (Fa), pRotein assembLIeS that couplE coNtractile actOmyOsin buNdLes TO the pLasMa memBrane aND transMit force gEnERated bY The cytoSKElEton To the ECM. Although iT HaS Been

iffer environments) is tho ught to un derly  ma nybi olog ical  processes inc l udin g angiogenesis, neurog enesi sa nd c a nc er me tastasi s .E C M s ti ff nes ss en singisachieve d by integ rin -m ediated foca l  a dhesions ( FA) , protein as sem bliesth atc ouple  co ntrac tile a c tomyos in bundle st o thep lasma m e m br aneand transmit forc e  g e nerated by the  cytos ke l et o n  to  th e ECM. Alt ho ugh i t  has be e n

iffer environments)_is thought_to underly many biological_processes including_angiogenesis,_neurogenesis and_cancer_metastasis. ECM stiffness_sensing is achieved_by integrin-mediated focal adhesions_(FA), protein assemblies_that_couple contractile actomyosin bundles to the plasma membrane and transmit force generated by the_cytoskeleton_to the_ECM._Although_it has been


the first dose was reduced 10-15 fold compared to the response in control animals that were HPIV3-nave. However, the serum antibody responses following the second dose were indistinguishable in HPIV3-immune versus HPIV3-naive animals. Thus, an HPIV3-based vector was substantially immun

the first dose was reduced 10 - 15 fold compare to the reception in control animals that were HPIV3 - nave. However, the serum antibody responses postdate the second dose were indistinguishable in HPIV3 - immune versus HPIV3 - primitive animals. Thus, an HPIV3 - establish vector was substantially immun

thf first dose was reduced 10-15 fold compared to the respohse in cuntrol animals that were HPIT3-navw. Howtner, the serum antibodh responsvs followung uhe second dose wxde indistingujdhabnx in HPIV3-immune versus HPHV3-naive animalv. Ghbs, an HPIV3-based vector was substantiwlly imkuj

the first dose was reduced 10-15 fold the in control that were HPIV3-nave. following second dose were in HPIV3-immune versus animals. Thus, an HPIV3-based vector was immun

the first dose was reduced 10-15 folD compared tO the rEspOnsE iN conTrol Animals that werE hPIV3-Nave. However, the serum antIbody ReSPonsES fOllowIng the sECoND DosE wErE inDiSTiNguisHabLe in HPIv3-immune verSus hPiV3-naive animaLS. THus, an HPIV3-bAseD vector was suBstAntialLy ImmUN

the first dose was reduce d 10-15 fo ld co mpa red  t o th e re sponse in cont r ol a nimals that were HPIV3 -nave .H owev e r,  theserum a n ti b o dyre sp ons es fo llowi ngthe sec ond dose w ere  i ndistinguish a bl e in HPIV3 -im mune versusHPI V3-nai ve  an i mals.  Th us, a n HPIV 3 -based  vector w as substa n tiallyi m mu n

the_first dose_was reduced 10-15 fold_compared to_the_response in_control_animals that were_HPIV3-nave. However, the_serum antibody responses following_the second dose_were_indistinguishable in HPIV3-immune versus HPIV3-naive animals. Thus, an HPIV3-based vector was substantially immun


, late TGF-beta signature accurately predicted liver metastasis and discriminated HCC cell lines by degree of invasiveness. In addition, the TGF-beta gene expression signature possessed a predictive value for tumors other than HCC. These data demonstrate the clinical significance of the genes embedded in the TGF-beta expression signature for the molecular classification of HCC

, late TGF - beta signature accurately predicted liver metastasis and discriminated HCC cellular telephone line by degree of invasiveness. In addition, the TGF - beta gene formula key signature possessed a predictive value for tumors early than HCC. These data demonstrate the clinical significance of the gene implant in the TGF - beta expression key signature for the molecular classification of HCC

, lahe TGF-beta signature accmrately predicteb liver metasfasis ana discriminated HCC cell linxs bt degeee of invasiveness. In addition, the TGF-veta tene expression sinuaturs posvxssed a predictlve value fmr tumors othes ghcn HCC. These data demonstrate the clynical xihnificance of jhe gtnef emgvdbed in the TGF-beta expression sjgnaturt for the moleculat classification of HCC

, late TGF-beta signature accurately predicted liver discriminated cell lines degree of invasiveness. expression possessed a predictive for tumors other HCC. These data demonstrate the clinical of the genes embedded in the TGF-beta expression signature for the molecular classification HCC

, late TGF-beta signature accurAtely prediCted lIveR meTaStasIs anD discriminated hcC ceLl lines by degree of invasIveneSs. iN addITiOn, the tGF-beta GEnE EXprEsSiOn sIgNAtUre poSseSsed a prEdictive vaLue FoR tumors other THaN HCC. These dAta Demonstrate tHe cLinicaL sIgnIFicanCe oF the gEnes emBEdded iN the TGF-beTa EXpressIOn signaTURe For tHe molecular classiFIcATion of HCC

, late TGF-beta signatureaccurately  pred ict edli vermeta stasis and dis c rimi nated HCC cell lines b y deg re e  ofi nv asive ness. I n  a d d iti on ,the  T G F- betagen e expre ssion sign atu re  possessed a pr edictive v alu e for tumors  ot her th an  HC C . The sedatademons t rate t he clinic al signif i cance o f th e ge nes embedded in t h eT GF-beta expres sion s ig n at u r e f orthe molecu la r cla s sificat i on o f  HC C

, late_TGF-beta signature_accurately predicted liver metastasis_and discriminated_HCC_cell lines_by_degree of invasiveness._In addition, the_TGF-beta gene expression signature_possessed a predictive_value_for tumors other than HCC. These data demonstrate the clinical significance of the genes_embedded_in the_TGF-beta_expression_signature for the molecular classification_of HCC


intact target plasmid and found ninety-five percent occurred within a 160 nt region in the ade6 promoter. To determine which sequences of Ade6 were required for efficient integration, we created a series of 10 deletions within the target plasmid. This analysis revealed that the 160 nt region of the promoter was the only sequence that was

intact target plasmid and found ninety - five percent occur within a 160 national trust region in the ade6 promoter. To settle which sequence of Ade6 were required for efficient consolidation, we created a series of 10 deletions within the prey plasmid. This analysis revealed that the 160 national trust region of the promoter was the only sequence that was

inhact target plasmid and nound ninety-five percenv occurded withkn a 160 nt region in the ade6 pcomorer. Ti determine which sequdnces of Wde6 were reqnired for efficixht intenxatioh, we erxated a series pf 10 deletimns within the txryet plasmid. This analysis revealed trat the 160 jt region of tre pgoioted was the only sequence that was

intact target plasmid and found ninety-five percent a nt region the ade6 promoter. Ade6 required for efficient we created a of 10 deletions within the target This analysis revealed that the 160 nt region of the promoter was the sequence that was

intact target plasmid and fouNd ninety-fiVe perCenT ocCuRred WithIn a 160 nt region in tHE ade6 Promoter. To determine whiCh seqUeNCes oF adE6 were RequireD FoR EFfiCiEnT inTeGRaTion, wE crEated a sEries of 10 delEtiOnS within the taRGeT plasmid. ThIs aNalysis reveaLed That thE 160 nT reGIon of The PromoTer was THe only Sequence tHaT Was

intact target plasmid and  found nin ety-f ive  pe rc entoccu rred within a1 60 n t region in the ade6 p romot er .  Tod et ermin e which se q u enc es  o f A de 6  w ere r equ ired fo r efficien t i nt egration, we cr eated a se rie s of 10 dele tio ns wit hi n t h e tar get  plas mid. T h is ana lysis rev ea l ed tha t  the 16 0 nt  reg ion of the promot e rw as the only se quence  t h at w as

intact_target plasmid_and found ninety-five percent_occurred within_a_160 nt_region_in the ade6_promoter. To determine_which sequences of Ade6_were required for_efficient_integration, we created a series of 10 deletions within the target plasmid. This analysis_revealed_that the_160_nt_region of the promoter was_the only sequence that was


scanning microscope specifically designed for studying protein-protein interactions in living cells. The microscope is a fully functional laser scanning two-photon microscope, with the additional capabilities of measuring florescent emission spectra (spectral imaging), fluorescent lifetime decays (FLIM), and fluorescent anisotropy lifetime decays (rFLIM). These added capabilities make it

scanning microscope specifically designed for studying protein - protein interactions in support cell. The microscope is a fully functional laser scanning two - photon microscope, with the extra capabilities of measuring florescent emission spectra (apparitional imagination), fluorescent lifetime decays (FLIM), and fluorescent anisotropy lifetime decays (rFLIM). These added capability make it

scwnning microscope speciflcally designed yir stuvying pdotein-prutein interactions in living cwlls. Uke microscope is a fjlly funcnional lawer wxanning two-photon microsdlpe, civh the additionsl capabilhties of measusivg florescent emission spectra (spectrwl imagonh), fluorescent jifeuimq dedays (FLIM), and fluorescent anisotroly lifeuime decays (rFLIM). Yhese added capabilities mwke lt

scanning microscope specifically designed for studying protein-protein living The microscope a fully functional the capabilities of measuring emission spectra (spectral fluorescent lifetime decays (FLIM), and fluorescent lifetime decays (rFLIM). These added capabilities make it

scanning microscope specifiCally desigNed foR stUdyInG proTein-Protein interacTIons In living cells. The microsCope iS a FUlly FUnCtionAl laser SCaNNIng TwO-pHotOn MIcRoscoPe, wIth the aDditional cApaBiLities of measURiNg floresceNt eMission spectRa (sPectraL iMagINg), fluOreScent LifetiME decayS (FLIM), and fLuORescenT AnisotrOPY lIfetIme decays (rFLIM). TheSE aDDed capabilitieS make iT

scanning microscope speci fically de signe d f orst udyi ng p rotein-protein inte ractions in living cel ls. T he micr o sc ope i s a ful l yf u nct io na l l as e rscann ing  two-ph oton micro sco pe , with the a d di tional cap abi lities of me asu ring f lo res c ent e mis sionspectr a  (spec tral imag in g ), flu o rescent l if etim e decays (FLIM),a nd fluorescent an isotro py li f e tim e d ecays (rFL IM ). Th e se adde d  c a p a bil i ties make it

scanning_microscope specifically_designed for studying protein-protein_interactions in_living_cells. The_microscope_is a fully_functional laser scanning_two-photon microscope, with the_additional capabilities of_measuring_florescent emission spectra (spectral imaging), fluorescent lifetime decays (FLIM), and fluorescent anisotropy lifetime decays_(rFLIM)._These added_capabilities_make_it


, HIV-2 and SIV encoded Vpx counteracts the antiviral effects of SAMHD1 by targeting the protein for proteasomal degradation. In FY13 we initiated a new SAMHD1-related project that addresses the role of post-translational modifications in SAMHD1 for its antiviral activity. We used 32

, HIV-2 and SIV encoded Vpx counteracts the antiviral effects of SAMHD1 by targeting the protein for proteasomal degradation. In FY13 we initiate a newfangled SAMHD1 - related project that addresses the character of post - translational modifications in SAMHD1 for its antiviral activity. We use 32

, HIG-2 and SIV encoded Vpx comnteracts the anjiciral xffects of SAMHA1 by targeting the protein flr prottcsomal degradation. Iv FY13 we ijitiated a ntw SAMHD1-related pckject tmct adsvessev the role of ppst-transladional modificdtkous in SAMHD1 for its antiviral activiey. We uxef 32

, HIV-2 and SIV encoded Vpx counteracts effects SAMHD1 by the protein for initiated new SAMHD1-related project addresses the role post-translational modifications in SAMHD1 for its activity. We used 32

, HIV-2 and SIV encoded Vpx counteRacts the anTivirAl eFfeCtS of SaMHD1 By targeting the PRoteIn for proteasomal degradAtion. in fy13 we iNItIated A new SAMhd1-rELAteD pRoJecT tHAt AddreSseS the rolE of post-traNslAtIonal modificATiOns in SAMHD1 For Its antiviral ActIvity. WE uSed 32

, HIV-2 and SIV encoded Vp x countera cts t heant iv iral  eff ects of SAMHD1 by t argeting the protein f or pr ot e asom a ldegra dation. In F Y13  w eini ti a te d a n ewSAMHD1- related pr oje ct  that addres s es  the roleofpost-transla tio nal mo di fic a tions  in  SAMH D1 for its an tiviral a ct i vity.W e used3 2

, HIV-2_and SIV_encoded Vpx counteracts the_antiviral effects_of_SAMHD1 by_targeting_the protein for_proteasomal degradation. In_FY13 we initiated a_new SAMHD1-related project_that_addresses the role of post-translational modifications in SAMHD1 for its antiviral activity. We used_32


(Dlc1neo) were obtained, and the frt-flanked neo cassette was removed by breeding with Flpe transgenic mice, to generate the floxed Dlc1 allele (Dlc1fl) with single loxP sites on both sides of exon 4. Both the Dlc1neo/

(Dlc1neo) were obtained, and the frt - flanked neo cassette was removed by breeding with Flpe transgenic mouse, to render the floxed Dlc1 allele (Dlc1fl) with single loxP sites on both sides of exon 4. Both the Dlc1neo/

(Dlf1neo) were obtained, and tme frt-flanked neo cassevte was removed by breeding with Flpe transjenix mict, to generate the fuoxed Dlc1 allele (Elc1fo) with single loxP sites kk botk wides of exon 4. Both the Glc1neo/

(Dlc1neo) were obtained, and the frt-flanked neo removed breeding with transgenic mice, to (Dlc1fl) single loxP sites both sides of 4. Both the Dlc1neo/

(Dlc1neo) were obtained, and the fRt-flanked nEo casSetTe wAs RemoVed bY breeding with FLPe trAnsgenic mice, to generate The flOxED Dlc1 ALlEle (DlC1fl) with SInGLE loXP SiTes On BOtH sideS of Exon 4. BotH the Dlc1neo/

(Dlc1neo) were obtained,and the fr t-fla nke d n eo  cas sett e was removedb y br eeding with Flpe trans genic  m i ce,t ogener ate the fl o x edDl c1  al le l e(Dlc1 fl)  with s ingle loxP  si te s on both si d es  of exon 4 . B oth the Dlc1 neo /

(Dlc1neo)_were obtained,_and the frt-flanked neo_cassette was_removed_by breeding_with_Flpe transgenic mice,_to generate the_floxed Dlc1 allele (Dlc1fl)_with single loxP_sites_on both sides of exon 4. Both the Dlc1neo/


reproduce those observed in a subset of human HCC with poor prognosis. Our goal is to achieve a comprehensive characterization of the multiple-step tumor progression by profiling gene expression from the early to late stages of HCC development. Significantly, gene expression data were highly consistent with the c-Myc/Tgf&#9

reproduce those observed in a subset of human HCC with poor prognosis. Our goal is to achieve a comprehensive word picture of the multiple - gradation tumor progression by profiling gene expression from the early to former stages of HCC development. importantly, gene expression data were highly consistent with the c - Myc / Tgf&#9

reoroduce those observed ik a subset of human HCC with loor proenosis. Our goal is to achievx a xomprtkensive characterizagion of tje multiple-suep tumor progression by ixofiljkg geue expression frpm the earny to late stacer lf HCC development. Significantly, gege exprrsdion data were hignjy cknsistent with the c-Myc/Tgf&#9

reproduce those observed in a subset of with prognosis. Our is to achieve multiple-step progression by profiling expression from the to late stages of HCC development. gene expression data were highly consistent with the c-Myc/Tgf&#9

reproduce those observed in a Subset of huMan HCc wiTh pOoR proGnosIs. Our goal is to aCHievE a comprehensive charactErizaTiON of tHE mUltipLe-step tUMoR PRogReSsIon By PRoFilinG geNe expreSsion from tHe eArLy to late stagES oF HCC develoPmeNt. SignificanTly, Gene exPrEssIOn datA weRe higHly conSIstent With the c-MYc/tGf&#9

reproduce those observedin a subse t ofhum anHC C wi th p oor prognosis. Ourgoal is to achieve a c ompre he n sive ch aract erizati o no f  th emu lti pl e -s tep t umo r progr ession bypro fi ling gene ex p re ssion from  th e early to l ate  stage sofH CC de vel opmen t. Sig n ifican tly, gene  e x pressi o n dataw e re  hig hly consistent wi t ht he c-Myc/Tgf&a mp;#9

reproduce_those observed_in a subset of_human HCC_with_poor prognosis._Our_goal is to_achieve a comprehensive_characterization of the multiple-step_tumor progression by_profiling_gene expression from the early to late stages of HCC development. Significantly, gene expression_data_were highly_consistent_with_the c-Myc/Tgf&#9


of cargo release from single insulin-containing dense-core vesicles in cultured beta cells. Our hypothesis is that these proteins (dynamin, syndapin, amphiphysin, and endophilin) regulate the dilation of the fusion pore to control the amount of cargo released during single exocytic fusion events in cells

of cargo release from single insulin - containing dense - core vesicle in civilized beta cellular telephone. Our hypothesis is that these proteins (dynamin, syndapin, amphiphysin, and endophilin) baffle the dilation of the fusion pore to master the measure of cargo released during single exocytic fusion events in cell

of cargo release from singue insulin-contanbing dxnse-cors vesiclds in cultured beta cells. Ouc hypothewis is that these protdins (dynalin, syndqpin, qmphiphysii, and endophiljk) regblete the dilatiok of the fuvion pore to cmngrll the amount of cargo released duryng sinblf exocytic fusyon tvegts jn cells

of cargo release from single insulin-containing dense-core cultured cells. Our is that these endophilin) the dilation of fusion pore to the amount of cargo released during exocytic fusion events in cells

of cargo release from single iNsulin-contAininG deNse-CoRe veSiclEs in cultured beTA celLs. Our hypothesis is that tHese pRoTEins (DYnAmin, sYndapin, AMpHIPhySiN, aNd eNdOPhIlin) rEguLate the Dilation of The FuSion pore to coNTrOl the amounT of Cargo releaseD duRing siNgLe eXOcytiC fuSion eVents iN Cells

of cargo release from sin gle insuli n-con tai nin gdens e-co re vesicles in cult ured beta cells. Our h ypoth es i s is th at th ese pro t ei n s  (d yn am in,  s y nd apin,  am phiphys in, and en dop hi lin) regulat e  t he dilatio n o f the fusion  po re toco ntr o l the  am ountof car g o rele ased duri ng single exocyti c fu sion  events in cells

of_cargo release_from single insulin-containing dense-core_vesicles in_cultured_beta cells._Our_hypothesis is that_these proteins (dynamin,_syndapin, amphiphysin, and endophilin)_regulate the dilation_of_the fusion pore to control the amount of cargo released during single exocytic fusion_events_in cells


specific receptors to evoke the effector activities of mature T cells in the body, as well as regulate their growth, inactivation, differentiation, or death. In particular, we want to understand in molecular detail the protein-protein interactions that turn recognition of antigen by T cells into signals guiding the normal survival and effector functions of these

specific receptors to evoke the effector activities of ripe metric ton cells in the body, as well as baffle their growth, inactivation, specialization, or death. In particular, we want to sympathize in molecular contingent the protein - protein interactions that turn realization of antigen by T cells into signal guide the normal survival and effector function of these

spfcific receptors to evokt the effector acjicities of mafure T cdlls in the body, as well as ceguoate ukeir growth, inactivagion, diffvrentiatiin, oc death. In partirhlar, we want fl unbecstand in molecolar detail dhe protein-prodekn interactions that turn recognition of antogfn by T cells ynto fignzls guiding the normal survival ans effecuor functions of tnese

specific receptors to evoke the effector activities T in the as well as or In particular, we to understand in detail the protein-protein interactions that turn of antigen by T cells into signals guiding the normal survival and effector of these

specific receptors to evoke tHe effector ActivItiEs oF mAturE T ceLls in the body, as WEll aS regulate their growth, inActivAtIOn, diFFeRentiAtion, or DEaTH. in pArTiCulAr, WE wAnt to UndErstand In moleculaR deTaIl the protein-PRoTein interaCtiOns that turn rEcoGnitioN oF anTIgen bY T cElls iNto sigNAls guiDing the noRmAL surviVAl and efFECtOr fuNctions of these

specific receptors to evo ke the eff ector  ac tiv it iesof m ature T cellsi n th e body, as well as reg ulate  t h eirg ro wth,inactiv a ti o n , d if fe ren ti a ti on, o r d eath. I n particul ar,  w e want to un d er stand in m ole cular detail  th e prot ei n-p r otein  in terac tionst hat tu rn recogn it i on ofa ntigenb y  T  cel ls into signals g u id i ng the normalsurviv al an d eff ect or functio ns  of t h ese

specific_receptors to_evoke the effector activities_of mature_T_cells in_the_body, as well_as regulate their_growth, inactivation, differentiation, or_death. In particular,_we_want to understand in molecular detail the protein-protein interactions that turn recognition of antigen_by_T cells_into_signals_guiding the normal survival and_effector functions of these


been referred with autoinflammatory disorders presenting early in life, who have clinical similarities with NOMID and DIRA patients but can clinically not be classified. We are using a genetic approach of whole genome sequencing in collaboration with Dr. Kastners group, an immunologic approach to characterize the immune deviation and a treatment approach

been referred with autoinflammatory disorders presenting early in biography, who take clinical similarities with NOMID and DIRA patients but can clinically not be classified. We are use a genetic approach of unharmed genome sequencing in collaboration with Dr. Kastners group, an immunologic access to characterize the immune deviation and a discussion approach

befn referred with autoinfuammatory disorbwrs prxsentinf early kn life, who have clinical silioaritues with NOMID and DIRX patientd but cab clmnically not be rmassificb. We zve usnnj a genetic apptoach of whone genome sequanziug in collaboration with Dr. Kastners group, sn immunologic akproabh to dharacterize the immune deviation znd a tgeatment approach

been referred with autoinflammatory disorders presenting early who clinical similarities NOMID and DIRA be We are using genetic approach of genome sequencing in collaboration with Dr. group, an immunologic approach to characterize the immune deviation and a treatment approach

been referred with autoinflaMmatory disOrderS prEseNtIng eArly In life, who have cLInicAl similarities with NOMId and DiRa PatiENtS but cAn cliniCAlLY Not Be ClAssIfIEd. we are UsiNg a geneTic approacH of WhOle genome seqUEnCing in collAboRation with Dr. kasTners gRoUp, aN ImmunOloGic apProach TO charaCterize thE iMMune deVIation aND A tReatMent approach

been referred with autoin flammatory  diso rde rspr esen ting  early in life ,  who  have clinical similar ities  w i th N O MI D and  DIRA p a ti e n tsbu tcan  c l in icall y n ot be c lassified.  We  a re using a g e ne tic approa chof whole gen ome  seque nc ing in co lla borat ion wi t h Dr.Kastnersgr o up, an immunol o g ic  app roach to characte r iz e  the immune de viatio na nd a  tr eat ment appro ac h

been_referred with_autoinflammatory disorders presenting early_in life,_who_have clinical_similarities_with NOMID and_DIRA patients but_can clinically not be_classified. We are_using_a genetic approach of whole genome sequencing in collaboration with Dr. Kastners group, an_immunologic_approach to_characterize_the_immune deviation and a treatment_approach


percent of the bulk cell population at >2 weeks post-electroporation. Site-specific transgene integration was confirmed by PCR analysis of bulk cell gDNA using reporter- and flanking gene-specific primer pairs. To estimate the frequency of transgene integration, CD34+ cells were plated in a CFU assay and transplanted into NS

percent of the bulk cell population at > 2 weeks post - electroporation. web site - specific transgene consolidation was confirm by PCR analysis of bulk cell gDNA use reporter- and flanking gene - specific primer pair. To calculate the frequency of transgene integration, CD34 + cells were plated in a CFU assay and transplanted into NS

pegcent of the bulk cell pupulation at >2 wgejs posv-electrkporatiov. Site-specific transgene intxgrarion qas confirmed by PCR avalysis ov bulk cwll jDNA using reporvsr- and njankjkg geue-wpecific primet pairs. To evtimate the fraqjeucy of transgene integration, CD34+ cellf were llwted in a CFU wssaj wnd fgaksplanted into NS

percent of the bulk cell population at post-electroporation. transgene integration confirmed by PCR using and flanking gene-specific pairs. To estimate frequency of transgene integration, CD34+ cells plated in a CFU assay and transplanted into NS

percent of the bulk cell populAtion at >2 weeKs posT-elEctRoPoraTion. site-specific trANsgeNe integration was confirMed by pCr AnalYSiS of buLk cell gdnA USIng RePoRteR- aND fLankiNg gEne-specIfic primer PaiRs. to estimate thE FrEquency of tRanSgene integraTioN, CD34+ celLs WerE PlateD in A CFU aSsay anD TranspLanted intO Ns

percent of the bulk cellpopulation  at > 2 w eek spost -ele ctroporation.S ite- specific transgene int egrat io n  was co nfirm ed by P C Ra n aly si sofbu l kcellgDN A using  reporter-  an dflanking gen e -s pecific pr ime r pairs. Toest imateth e f r equen cyof tr ansgen e  integ ration, C D3 4 + cell s  were p l a te d in  a CFU assay andt ra n splanted intoNS

percent_of the_bulk cell population at_>2 weeks_post-electroporation._Site-specific transgene_integration_was confirmed by_PCR analysis of_bulk cell gDNA using_reporter- and flanking_gene-specific_primer pairs. To estimate the frequency of transgene integration, CD34+ cells were plated in_a_CFU assay_and_transplanted_into NS


, and skeletal, neurological and renal anomalies. These additional features may be the key to determining the developmental function of genes implicated in this spectrum of disorders. Our clinical protocol (http://gp.nichd.nih.gov), which is a multicenter study in collaboration with the REU at MGH has identified a

, and skeletal, neurological and renal anomalies. These additional feature may be the keystone to determining the developmental function of genes entail in this spectrum of disorders. Our clinical protocol (http://gp.nichd.nih.gov), which is a multicenter study in collaboration with the REU at MGH has identify a

, anf skeletal, neurological xnd renal anomalies. Thxse addjtional weatures may be the key to dxterninint the developmental fuvction of genes inplirated in this spxdtrum on diskvders. Iur clinical ptotocol (http://cp.nichd.nih.gov), fhkck is a multicenter study in collaborwtion wotj the REU at MDH hsf idsntified a

, and skeletal, neurological and renal anomalies. features be the to determining the in spectrum of disorders. clinical protocol (http://gp.nichd.nih.gov), is a multicenter study in collaboration the REU at MGH has identified a

, and skeletal, neurological anD renal anomAlies. theSe aDdItioNal fEatures may be thE Key tO determining the developMentaL fUNctiON oF geneS implicATeD IN thIs SpEctRuM Of DisorDerS. Our cliNical protoCol (HtTp://gp.nichd.nih.GOv), Which is a muLtiCenter study iN coLlaborAtIon WIth thE REu at MGh has idENtifieD a

, and skeletal, neurologic al and ren al an oma lie s.  The se a dditional feat u resmay be the key to dete rmini ng thed ev elopm ental f u nc t i onof  g ene si mp licat edin this  spectrumofdi sorders. Our cl inical pro toc ol (http://g p.n ichd.n ih .go v ), wh ich  is a  multi c enterstudy inco l labora t ion wit h th e RE U at MGH has iden t if i ed a

, and_skeletal, neurological_and renal anomalies. These_additional features_may_be the_key_to determining the_developmental function of_genes implicated in this_spectrum of disorders._Our_clinical protocol (http://gp.nichd.nih.gov), which is a multicenter study in collaboration with the REU at_MGH_has identified_a


shown that FCR3 parasitized red cells expressing VAR2CSA can be efficiently opsonized by a human monocytic cell line. We have developed a novel procedure testing the ability of various domains to inhibit opsonization by these antibodies. We have thus identified several domains that are important for recognition by anti-

shown that FCR3 parasitized red cells expressing VAR2CSA can be efficiently opsonize by a human monocytic cellular telephone occupation. We have developed a fresh procedure test the ability of various world to suppress opsonization by these antibodies. We have thus identified several domains that are authoritative for recognition by anti-

shlwn that FCR3 parasitized red cells exprgswing VER2CSA czn be efwiciently opsonized by a humen minocyupc cell line. We have aeveloped a novel proredure testing tis abilibv of bwriobs domains to inmibit opsonhzation by theve autibodies. We have thus identified sederal dpmwins that are ympogtwnt ror recognition by anti-

shown that FCR3 parasitized red cells expressing be opsonized by human monocytic cell novel testing the ability various domains to opsonization by these antibodies. We have identified several domains that are important for recognition by anti-

shown that FCR3 parasitized reD cells exprEssinG VAr2CSa cAn be EffiCiently opsonizED by a Human monocytic cell line. we havE dEVeloPEd A noveL procedURe TEStiNg ThE abIlITy Of varIouS domainS to inhibit OpsOnIzation by theSE aNtibodies. WE haVe thus identiFieD severAl DomAIns thAt aRe impOrtant FOr recoGnition by AnTI-

shown that FCR3 parasitiz ed red cel ls ex pre ssi ng  VAR 2CSA  can be effici e ntly  opsonized by a humanmonoc yt i c ce l lline.  We hav e  d e v elo pe da n ov e lproce dur e testi ng the abi lit yof various d o ma ins to inh ibi t opsonizati onby the se  an t ibodi es.  We h ave th u s iden tified se ve r al dom a ins tha t ar e im portant for recog n it i on by anti-

shown_that FCR3_parasitized red cells expressing_VAR2CSA can_be_efficiently opsonized_by_a human monocytic_cell line. We_have developed a novel_procedure testing the_ability_of various domains to inhibit opsonization by these antibodies. We have thus identified several_domains_that are_important_for_recognition by anti-


2287, 2002);c) addressed the mechanistic differences between the CCR5 and CXCR4 receptors in the agonist-driven receptor signaling, and trafficking (Mol Biol of Cell 14, 3305, 2003), d) showed that in primary leukocytes, agonist driven cell migration and receptor internalization are separable processes (J.

2287, 2002);c) addressed the mechanistic differences between the CCR5 and CXCR4 receptors in the agonist - drive sense organ signaling, and trafficking (Mol Biol of Cell 14, 3305, 2003), d) testify that in elementary leukocytes, agonist driven cellular telephone migration and receptor internalization are separable processes (J.

2287, 2002);c) addressed the mechanistlc differences bgtqeen tie CCR5 znd CXCR4 receptors in the agonist-driten eecepujr signaling, and tfaffickinh (Mol Biil oh Cell 14, 3305, 2003), d) showxs that lu prijwry nxukocytes, agonixt driven well migration avd receptor internalization are separwble prpcfsses (J.

2287, 2002);c) addressed the mechanistic differences between and receptors in agonist-driven receptor signaling, Cell 3305, 2003), d) that in primary agonist driven cell migration and receptor are separable processes (J.

2287, 2002);c) addressed the mechanistic dIfferences BetweEn tHe CcR5 And CxCR4 rEceptors in the aGOnisT-driven receptor signaliNg, and TrAFficKInG (Mol BIol of CeLL 14, 3305, 2003), d) SHOweD tHaT in PrIMaRy leuKocYtes, agoNist driven CelL mIgration and rECePtor internAliZation are sepAraBle proCeSseS (j.

2287, 2002);c) addressedthe mechan istic  di ffe re nces  bet ween the CCR5a nd C XCR4 receptors in theagoni st - driv e nrecep tor sig n al i n g,an dtra ff i ck ing ( Mol  Biol o f Cell 14,  33 05 , 2003), d)s ho wed that i n p rimary leuko cyt es, ag on ist drive n c ell m igrati o n andreceptorin t ernali z ation a r e  s epar able processes (J .

2287,_2002);c) addressed_the mechanistic differences between_the CCR5_and_CXCR4 receptors_in_the agonist-driven receptor_signaling, and trafficking_(Mol Biol of Cell_14, 3305, 2003),_d)_showed that in primary leukocytes, agonist driven cell migration and receptor internalization are separable_processes_(J.


rontal cortical network oscillations. We found that modification of CA2 activity increased or decreased hippocampal and prefrontal cortical low-gamma oscillations with excitatory and inhibitory DREADDs, respectively, and that CA2 activity inversely modulated hippocampal ripple oscillations. These findings support a role for CA2 in low-gamma generation and ripple modulation

rontal cortical network oscillations. We found that modification of CA2 natural process increase or decreased hippocampal and prefrontal cortical low - gamma oscillation with excitatory and inhibitory DREADDs, respectively, and that CA2 activity inversely modulated hippocampal ripple oscillation. These findings support a function for CA2 in low - gamma genesis and ripple modulation

ronhal cortical network osclllations. We fouue that modifjcation uf CA2 activity increased or vecrwased hippocampal and prefruntal cornical low-tamme oscillations wmfh excibctory wnd nniibitory DREADDx, respectieely, and that WA2 aetivity inversely modulated hippocam[al ripllf oscillations. Thexq fihdings support a role for CA2 in los-gamma jeneration and tipple modulation

rontal cortical network oscillations. We found that CA2 increased or hippocampal and prefrontal and DREADDs, respectively, and CA2 activity inversely hippocampal ripple oscillations. These findings support role for CA2 in low-gamma generation and ripple modulation

rontal cortical network osciLlations. We Found ThaT moDiFicaTion Of CA2 activity inCReasEd or decreased hippocampAl and PrEFronTAl CortiCal low-gAMmA OSciLlAtIonS wITh ExcitAtoRy and inHibitory DReADds, Respectively, ANd That CA2 actiVitY inversely moDulAted hiPpOcaMPal riPplE osciLlatioNS. These Findings sUpPOrt a roLE for CA2 iN LOw-GammA generation and ripPLe MOdulation

rontal cortical network os cillations . Wefou ndth at m odif ication of CA2 acti vity increased or decr eased  h i ppoc a mp al an d prefr o nt a l  co rt ic allo w -g ammaosc illatio ns with ex cit at ory and inhi b it ory DREADD s,respectively , a nd tha tCA2 activ ity  inve rselym odulat ed hippoc am p al rip p le osci l l at ions . These findingss up p ort a role for  CA2 i nl ow - g amm a g enerationan d rip p le modu l at i o n

rontal cortical_network oscillations._We found that modification_of CA2_activity_increased or_decreased_hippocampal and prefrontal_cortical low-gamma oscillations_with excitatory and inhibitory_DREADDs, respectively, and_that_CA2 activity inversely modulated hippocampal ripple oscillations. These findings support a role for CA2_in_low-gamma generation_and_ripple_modulation


solved this issue by establishing an internal quality assurance program for key reagents and centrally procuring them, qualifying them for use in the validated assay, and then making them available upon request via the DCTD Biomarker's website to the oncology research community. In addition, this website provides access to the validated assay S

solved this issue by establishing an internal timbre assurance platform for key reagents and centrally procure them, qualify them for use in the validated assay, and then making them available upon request via the DCTD Biomarker's web site to the oncology research community. In addition, this web site provide access to the validate assay S

sopved this issue by estabuishing an intetnql quanity aasurance program for key reagents anv cebtraloy procuring them, qualkfying thvm for usw in rhe validavsd assan, and bhen keking them avaikable upon request via tve DETD Biomarker's website to the oncolody resesrfh community. Ig adcytioh, this website provides access to fhe valpdated assay S

solved this issue by establishing an internal program key reagents centrally procuring them, the assay, and then them available upon via the DCTD Biomarker's website to oncology research community. In addition, this website provides access to the validated assay

solved this issue by establisHing an inteRnal qUalIty AsSuraNce pRogram for key reAGentS and centrally procuring Them, qUaLIfyiNG tHem foR use in tHE vALIdaTeD aSsaY, aND tHen maKinG them avAilable upoN reQuEst via the DCTd biOmarker's weBsiTe to the oncolOgy ResearCh ComMUnity. in aDditiOn, this WEbsite Provides aCcESs to thE ValidatED AsSay S

solved this issue by esta blishing a n int ern alqu alit y as surance progra m  for  key reagents and cent rally  p r ocur i ng  them , quali f yi n g  th em  f orus e  i n the  va lidated  assay, an d t he n making the m  a vailable u pon  request via  th e DCTD  B iom a rker' s w ebsit e to t h e onco logy rese ar c h comm u nity. I n ad diti on, this websitep ro v ides access to  the v al i da t e d a ssa y S

solved_this issue_by establishing an internal_quality assurance_program_for key_reagents_and centrally procuring_them, qualifying them_for use in the_validated assay, and_then_making them available upon request via the DCTD Biomarker's website to the oncology research_community._In addition,_this_website_provides access to the validated_assay S


been implicated in susceptibility to bipolar disorder. Frustrated in our attempts to confirm a published report that the vertebrate ortholog functions as a non-selective cation channel, we have been unable to ask whether channel function is affected by anesthetics. Instead, we have pursued the functional anatomy of channel's contribution to anesthetic sensitivity. The

been implicated in susceptibility to bipolar disorder. Frustrated in our attempts to confirm a publish reputation that the vertebrate ortholog functions as a non - selective cation channel, we have been unable to necessitate whether channel function is affected by anesthetic. rather, we have pursued the running human body of channel's contribution to anesthetic sensitivity. The

befn implicated in susceptlbility to bipolce disocder. Frhstrated in our attempts to confirm e puvlishtb report that the veftebrate lrtholog funrtions as a non-sxmective catioh chaunxl, we have been unable to ask whether cvavnzl function is affected by anesthetiss. Instraf, we have pursoed tnq fuhbtlonal anatomy of channel's contrigution uo anesthetic sensotivity. The

been implicated in susceptibility to bipolar disorder. our to confirm published report that a cation channel, we been unable to whether channel function is affected by Instead, we have pursued the functional anatomy of channel's contribution to anesthetic sensitivity.

been implicated in susceptibIlity to bipOlar dIsoRdeR. FRustRateD in our attempts TO conFirm a published report thAt the VeRTebrATe OrthoLog funcTIoNS As a NoN-sEleCtIVe CatioN chAnnel, we Have been unAblE tO ask whether cHAnNel functioN is Affected by anEstHetics. inSteAD, we haVe pUrsueD the fuNCtionaL anatomy oF cHAnnel's COntribuTIOn To anEsthetic sensitiviTY. THE

been implicated in suscep tibility t o bip ola r d is orde r. F rustrated in o u r at tempts to confirm a pu blish ed repo r tthatthe ver t eb r a teor th olo gf un ction s a s a non -selective  ca ti on channel,w ehave beenuna ble to ask w het her ch an nel funct ion  is a ffecte d  by an esthetics .I nstead ,  we hav e pu rsue d the functionala na t omy of channel 's con tr i bu t i ontoanesthetic  s ensit i vity. T h e

been_implicated in_susceptibility to bipolar disorder._Frustrated in_our_attempts to_confirm_a published report_that the vertebrate_ortholog functions as a_non-selective cation channel,_we_have been unable to ask whether channel function is affected by anesthetics. Instead, we_have_pursued the_functional_anatomy_of channel's contribution to anesthetic_sensitivity. The


45;phenotype described in our previous work. Starting from 3 wk of age, microarray analysis revealed upregulation of genes involved in DNA replication (Mcm6, Mcm7, Myc), DNA repair (Ddb2, Rad51l1) and translation (Eef1b2, Eef2, R

45;phenotype described in our previous work. Starting from 3 wk of old age, microarray psychoanalysis revealed upregulation of genes imply in DNA rejoinder (Mcm6, Mcm7, Myc), DNA repair (Ddb2, Rad51l1) and transformation (Eef1b2, Eef2, R

45;phejotype described in our krevious work. Stattung frmm 3 wk of age, oicroarray analysis revealed upreguoation of genes involvdd in DNA replicarion (Ncm6, Mcm7, Myr), DNA reicir (Dsn2, Rad51n1) and translatipn (Eef1b2, Eex2, R

45;phenotype described in our previous work. Starting wk age, microarray revealed upregulation of (Mcm6, Myc), DNA repair Rad51l1) and translation Eef2, R

45;phenotype described in our prEvious work. startIng FroM 3 wK of aGe, miCroarray analysIS revEaled upregulation of genEs invOlVEd in dnA RepliCation (MCM6, MCM7, myc), dNa rEpaIr (dDb2, rad51l1) aNd tRanslatIon (Eef1b2, Eef2, r

45;phenotype described inour previo us wo rk.  St ar ting  fro m 3 wk of age, micr oarray analysis reveal ed up re g ulat i on  of g enes in v ol v e d i nDN A r ep l ic ation  (M cm6, Mc m7, Myc),DNA  r epair (Ddb2, Ra d51l1) and  tr anslation (E ef1 b2, Ee f2 , R

45;phenotype described_in our_previous work. Starting from_3 wk_of_age, microarray_analysis_revealed upregulation of_genes involved in_DNA replication (Mcm6, Mcm7,_Myc), DNA repair_(Ddb2,_Rad51l1) and translation (Eef1b2, Eef2, R


de novo somatic mutations in an unbiased, genome-wide manner via high-throughput, whole-genome sequencing (WGS) of single-cell-derived HSPC clones. Importantly, to distinguish between naturally occurring spontaneous somatic mutation, potential cell culture-induced mutagenesis, and bona fide Cas9-mediated genetic alterations, the

de novo somatic mutations in an unbiased, genome - wide manner via eminent - throughput, solid - genome sequencing (WGS) of single - cell - derive HSPC clones. Importantly, to distinguish between naturally happen spontaneous somatic mutant, likely cell culture - induce mutagenesis, and bona fide Cas9 - mediated genetic alterations, the

de novo somatic mutations ln an unbiased, ggnime-widx manned via hieh-throughput, whole-genome seqnencung (WTS) of single-cell-derivea HSPC cllnes. Impirtaitly, to distingumah betwczn nafmrallv iccurring sponjaneous somadic mutation, pmtdncial cell culture-induced mutagenesis, and boma fide Cas9-mediajed gtneeic zlterations, the

de novo somatic mutations in an unbiased, via whole-genome sequencing of single-cell-derived HSPC naturally spontaneous somatic mutation, cell culture-induced mutagenesis, bona fide Cas9-mediated genetic alterations, the

de novo somatic mutations in aN unbiased, gEnome-WidE maNnEr viA higH-throughput, whoLE-genOme sequencing (WGS) of singLe-celL-dERiveD hSpC cloNes. ImpoRTaNTLy, tO dIsTinGuISh BetweEn nAturallY occurring SpoNtAneous somatiC MuTation, poteNtiAl cell culturE-inDuced mUtAgeNEsis, aNd bOna fiDe Cas9-mEDiated Genetic alTeRAtions, THe

de novo somatic mutations  in an unb iased , g eno me -wid e ma nner via high- t hrou ghput, whole-genome se quenc in g  (WG S )of si ngle-ce l l- d e riv ed  H SPC  c l on es. I mpo rtantly , to disti ngu is h between na t ur ally occur rin g spontaneou s s omatic  m uta t ion,pot entia l cell cultur e-induced  m u tagene s is, and b on a fi de Cas9-mediatedg en e tic alteration s, the

de_novo somatic_mutations in an unbiased,_genome-wide manner_via_high-throughput, whole-genome_sequencing_(WGS) of single-cell-derived_HSPC clones. Importantly,_to distinguish between naturally_occurring spontaneous somatic_mutation,_potential cell culture-induced mutagenesis, and bona fide Cas9-mediated genetic alterations, the


and FPR in monocytes, but not in PMNs, T cells or myelomonocytic cell lines. LPS treatment abolished polarized accumulation of F-actin and PIP3 in response to chemokine stimulation in monocytes but not in PMNs. TLR stimulation did not alter initial G-protein dependent signaling or the kinetics of endocytosis of

and FPR in monocytes, but not in PMNs, T cells or myelomonocytic cell occupation. LPS discussion abolished polarized accretion of F - actin and PIP3 in answer to chemokine stimulation in monocytes but not in PMNs. TLR foreplay did not alter initial G - protein dependent sign or the kinetics of endocytosis of

anf FPR in monocytes, but nut in PMNs, T cells or kyelomknocytic cell lines. LPS treatment abllushed polarized accumulatiov of F-actpn and PIP3 in eesponse to chemoklue stjlulaciin in monocytex but not hn PMNs. TLR sthmjlction did not alter initial G-protein dependrnh signaling or the hinefpcw of endocytosis of

and FPR in monocytes, but not in cells myelomonocytic cell LPS treatment abolished PIP3 response to chemokine in monocytes but in PMNs. TLR stimulation did not initial G-protein dependent signaling or the kinetics of endocytosis of

and FPR in monocytes, but not in pMNs, T cells Or myeLomOnoCyTic cEll lInes. LPS treatmeNT aboLished polarized accumulAtion Of f-ActiN AnD PIP3 iN responSE tO CHemOkInE stImULaTion iN moNocytes But not in PMns. TlR Stimulation dID nOt alter iniTiaL G-protein depEndEnt sigNaLinG Or the KinEtics Of endoCYtosis Of

and FPR in monocytes, but  not in PM Ns, T  ce lls  o r my elom onocytic celll ines . LPS treatment abolis hed p ol a rize d  a ccumu lationo fF - act in  a ndPI P 3in re spo nse tochemokinesti mu lation in mo n oc ytes but n otin PMNs. TLR  st imulat io n d i d not  al ter i nitial G-prot ein depen de n t sign a ling or t he  kin etics of endocyto s is of

and_FPR in_monocytes, but not in_PMNs, T_cells_or myelomonocytic_cell_lines. LPS treatment_abolished polarized accumulation_of F-actin and PIP3_in response to_chemokine_stimulation in monocytes but not in PMNs. TLR stimulation did not alter initial G-protein_dependent_signaling or_the_kinetics_of endocytosis of


its dysfunction has been implicated in many neuropsychiatric disorders. We used a J-edited, single voxel-based spectroscopy method to measure GABA + macromolecules in the anterior cingulate cortex and right frontal white matter. In fact, our method allowed us to quantify metabolite differences between gray and white matter. Reproducibility of

its dysfunction has been implicated in many neuropsychiatric disorders. We used a J - edited, individual voxel - free-base spectroscopy method to measure GABA + macromolecules in the anterior cingulate lens cortex and right frontal white topic. In fact, our method allowed us to quantify metabolite differences between grey and white matter. Reproducibility of

itd dysfunction has been ioplicated in maut neurmpsychjatric dksorders. We used a J-edited, smnglw voxtj-based spectroscoph method no measurw GAUA + macromoleculxa in thc antsvior eiigulate cortex snd right xrontal white kagtzr. In fact, our method allowed us to zuantifu letabolite difseremses gvtceen gray and white matter. Reprkducibinity of

its dysfunction has been implicated in many We a J-edited, voxel-based spectroscopy method in anterior cingulate cortex right frontal white In fact, our method allowed us quantify metabolite differences between gray and white matter. Reproducibility of

its dysfunction has been implIcated in maNy neuRopSycHiAtriC disOrders. We used a J-EDiteD, single voxel-based spectRoscoPy MEthoD To MeasuRe GABA + mACrOMOleCuLeS in ThE AnTerioR ciNgulate Cortex and rIghT fRontal white mATtEr. In fact, ouR meThod allowed uS to QuantiFy MetABolitE diFfereNces beTWeen grAy and whitE mATter. RePRoducibILItY of

its dysfunction has beenimplicated  in m any  ne ur opsy chia tric disorders .  Weused a J-edited, singl e vox el - base d  s pectr oscopym et h o d t ome asu re GA BA +mac romolec ules in th e a nt erior cingul a te  cortex an d r ight frontal  wh ite ma tt er. In fa ct,  ourmethod allowe d us to q ua n tify m e tabolit e di ffer ences between gra y  a n d white matter . Repr od u ci b i lit y o f

its_dysfunction has_been implicated in many_neuropsychiatric disorders._We_used a_J-edited,_single voxel-based spectroscopy_method to measure_GABA + macromolecules in_the anterior cingulate_cortex_and right frontal white matter. In fact, our method allowed us to quantify metabolite_differences_between gray_and_white_matter. Reproducibility of


candidate - PfRH5 - has been identified that elicits antibodies with very high levels of GIA activity. Binding of PfRH5 to the red cell protein basigin has been shown to be essential for merozoite invasion. We have tested antibodies to PfRH5 against parasites from different geographic locations and are evaluating

candidate - PfRH5 - has been identified that elicits antibodies with very high levels of GIA natural process. tie of PfRH5 to the red cellular telephone protein basigin has been shown to be substantive for merozoite invasion. We have tested antibodies to PfRH5 against parasite from different geographic locations and are measure

cajdidate - PfRH5 - has been ldentified that glucits entibodjes with very high levels of GIA actmvitt. Bineing of PfRH5 to the rea cell prltein bawigii has been shown to be essentizp fox nerozoite invaxion. We haee tested antitoaizs to PfRH5 against parasites from disferent gfographic locajions wnd zge evaluating

candidate - PfRH5 - has been identified antibodies very high of GIA activity. red protein basigin has shown to be for merozoite invasion. We have tested to PfRH5 against parasites from different geographic locations and are evaluating

candidate - PfRH5 - has been identIfied that eLicitS anTibOdIes wIth vEry high levels oF gIA aCtivity. Binding of PfRH5 to The reD cELl prOTeIn basIgin has BEeN SHowN tO bE esSeNTiAl for MerOzoite iNvasion. We hAve TeSted antibodiES tO PfRH5 againSt pArasites from DifFerent GeOgrAPhic lOcaTions And are EValuatIng

candidate - PfRH5 - has b een identi fiedtha t e li cits  ant ibodies with v e ry h igh levels of GIA acti vity.  B i ndin g  o f PfR H5 to t h er e d c el lpro te i nbasig inhas bee n shown to  be  e ssential for me rozoite in vas ion. We have  te sted a nt ibo d ies t o P fRH5agains t  paras ites from  d i fferen t  geogra p h ic  loc ations and are ev a lu a ting

candidate_- PfRH5_- has been identified_that elicits_antibodies_with very_high_levels of GIA_activity. Binding of_PfRH5 to the red_cell protein basigin_has_been shown to be essential for merozoite invasion. We have tested antibodies to PfRH5_against_parasites from_different_geographic_locations and are evaluating


directly with the PPT. This observation led us to test whether the mutations in the RNase H of Tf1 were defective for the cleavages on either side of the PPT that generate the primer, or the cleavage that removes the PPT after it has primed plus strand synthesis. Defects in the

directly with the PPT. This observation led us to test whether the mutation in the RNase H of Tf1 were bad for the cleavages on either side of the PPT that render the flat coat, or the cleavage that removes the PPT after it has primed plus fibril deduction. Defects in the

digectly with the PPT. This observation leb us to test shether ghe mutations in the RNase H od Tf1 qere defective for the cleavaged on eityer wude of the PPT thab gensvate chx primer, or the cleavage dhat removes tve P'T after it has primed plus strand srnthesix. Fefects in the

directly with the PPT. This observation led test the mutations the RNase H the on either side the PPT that the primer, or the cleavage that the PPT after it has primed plus strand synthesis. Defects in the

directly with the PPT. This obsErvation leD us to TesT whEtHer tHe muTations in the RNASe H oF Tf1 were defective for the CleavAgES on eIThEr sidE of the Ppt tHAT geNeRaTe tHe PRiMer, or The CleavagE that removEs tHe pPT after it haS PrImed plus stRanD synthesis. DeFecTs in thE

directly with the PPT. Th is observa tionled  us  t o te st w hether the mut a tion s in the RNase H of Tf 1 wer ed efec t iv e for  the cl e av a g eson  e ith er si de of  th e PPT t hat genera teth e primer, or th e cleavage  th at removes t hePPT af te r i t  haspri med p lus st r and sy nthesis.De f ects i n  the

directly_with the_PPT. This observation led_us to_test_whether the_mutations_in the RNase_H of Tf1_were defective for the_cleavages on either_side_of the PPT that generate the primer, or the cleavage that removes the PPT_after_it has_primed_plus_strand synthesis. Defects in the


of which changed in FA abundance in response to perturbation of myosinII activity, thus defining the myosinII-responsive FA proteome. FA abundance of 75% of proteins in the myosinII-responsive FA proteome were enhanced by contractility, including those involved in Rho-mediated FA maturation, stress fiber formation, and end

of which changed in FA abundance in response to perturbation of myosinII activity, therefore define the myosinII - responsive FA proteome. FA abundance of 75% of proteins in the myosinII - reactive FA proteome were enhanced by contractility, admit those involved in Rho - mediated FA growth, stress fiber constitution, and end

of which changed in FA abukdance in response to pxrturbafion of oyosinII activity, thus definmng rhe mtosinII-responsive FA pfoteome. FW abundabce id 75% of protxjns in bke mykdinIN-rxsponsive FA prpteome wera enhanced by wovtxactility, including those involved ig Rho-meciwted FA maturajion, xeresa fiber formation, and end

of which changed in FA abundance in perturbation myosinII activity, defining the myosinII-responsive 75% proteins in the FA proteome were by contractility, including those involved in FA maturation, stress fiber formation, and end

of which changed in FA abundanCe in responSe to pErtUrbAtIon oF myoSinII activity, tHUs deFining the myosinII-respoNsive fA PRoteOMe. fA abuNdance oF 75% Of PROteInS iN thE mYOsInII-rEspOnsive Fa proteome wEre EnHanced by contRAcTility, inclUdiNg those involVed In Rho-mEdIatED FA maTurAtion, Stress FIber foRmation, anD eND

of which changed in FA ab undance in  resp ons e t opert urba tion of myosin I I ac tivity, thus definingthe m yo s inII - re spons ive FAp ro t e ome .FA  ab un d an ce of  75 % of pr oteins inthe  m yosinII-resp o ns ive FA pro teo me were enha nce d by c on tra c tilit y,inclu ding t h ose in volved in  R h o-medi a ted FAm a tu rati on, stress fiberf or m ation, and end

of_which changed_in FA abundance in_response to_perturbation_of myosinII_activity,_thus defining the_myosinII-responsive FA proteome._FA abundance of 75%_of proteins in_the_myosinII-responsive FA proteome were enhanced by contractility, including those involved in Rho-mediated FA maturation,_stress_fiber formation,_and_end


one or more nonredundant roles in normal physiology. To gain additional insights into EpCAM function, we have generated mice with an EpCAM allele that can be conditionally deleted in a lineage-specific fashion. Working with Dr. Tessarollo and using recombineering, we developed a targeting vector that allowed l

one or more nonredundant roles in normal physiology. To gain additional insights into EpCAM affair, we have generate mice with an EpCAM allele that can be conditionally deleted in a lineage - specific manner. Working with Dr. Tessarollo and using recombineering, we developed a target vector that allow l

onf or more nonredundant rules in normal khtsiolojy. To gzin addigional insights into EpCAM fnnctuon, wt have generated mize with aj EpCAM qlleow that can be condlcionampy dzlxted in a lineane-specific xashion. Workinc dich Dr. Tessarollo and using recombineqring, wr feveloped a tatgetimd vednov that allowed l

one or more nonredundant roles in normal gain insights into function, we have allele can be conditionally in a lineage-specific Working with Dr. Tessarollo and using we developed a targeting vector that allowed l

one or more nonredundant roleS in normal pHysioLogY. To GaIn adDitiOnal insights inTO EpCaM function, we have generaTed miCe WIth aN epcAM alLele thaT CaN BE coNdItIonAlLY dEleteD in A lineagE-specific fAshIoN. Working with dR. TEssarollo aNd uSing recombinEerIng, we dEvEloPEd a taRgeTing vEctor tHAt alloWed l

one or more nonredundantroles in n ormal  ph ysi ol ogy.  Togain additiona l  ins ights into EpCAM funct ion,we have ge nerat ed mice wi t h  an  E pC AMal l el e tha t c an be c onditional lyde leted in a l i ne age-specif icfashion. Wor kin g with  D r.T essar oll o and  using recomb ineering,  w e  devel o ped a t a r ge ting  vector that allo w ed l

one_or more_nonredundant roles in normal_physiology. To_gain_additional insights_into_EpCAM function, we_have generated mice_with an EpCAM allele_that can be_conditionally_deleted in a lineage-specific fashion. Working with Dr. Tessarollo and using recombineering, we developed_a_targeting vector_that_allowed_l


ional pretreatment skin biopsy samples to post treatment skin biopsies and normal control tissue. 2. We studied a chondrocyte primary cell lines from a bone lesions from one patients with NOMID and in collaboration with Dr. Stratakis laboratory, their group identified that the cyclic adenosine-monophosphate (cAMP) signaling pathway is

ional pretreatment skin biopsy samples to post treatment hide biopsy and normal control tissue. 2. We studied a chondrocyte basal cellular telephone lines from a bone lesions from one patients with NOMID and in collaboration with Dr. Stratakis lab, their group identified that the cyclic adenosine - monophosphate (cAMP) sign pathway is

ionwl pretreatment skin bioksy samples to post treavment siin biopries and normal control tissne. 2. Qe stydied a chondrocyte prkmary celp lines drom q bone lesmkns from one lwtieutw with NOMID akd in collatoration with Gr. Scratakis laboratory, their group ideneified yhwt the cyclic wdenpfine-jonophosphate (cAMP) signaling pathwzy is

ional pretreatment skin biopsy samples to post biopsies normal control 2. We studied from bone lesions from patients with NOMID in collaboration with Dr. Stratakis laboratory, group identified that the cyclic adenosine-monophosphate (cAMP) signaling pathway is

ional pretreatment skin biopSy samples tO post TreAtmEnT skiN bioPsies and normal COntrOl tissue. 2. We studied a chonDrocyTe PRimaRY cEll liNes from A BoNE LesIoNs FroM oNE pAtienTs wIth NOMId and in collAboRaTion with Dr. StRAtAkis laboraTorY, their group iDenTified ThAt tHE cyclIc aDenosIne-monOPhosphAte (cAMP) siGnALing paTHway is

ional pretreatment skin bi opsy sampl es to  po sttr eatm entskin biopsiesa nd n ormal control tissue.2. We  s t udie d  a  chon drocyte pr i m ary  c el l l in e sfroma b one les ions fromone  p atients with NO MID and in  co llaborationwit h Dr.St rat a kis l abo rator y, the i r grou p identif ie d  thatt he cycl i c  a deno sine-monophosphat e  ( c AMP) signaling  pathw ay is

ional pretreatment_skin biopsy_samples to post treatment_skin biopsies_and_normal control_tissue._2. We studied_a chondrocyte primary_cell lines from a_bone lesions from_one_patients with NOMID and in collaboration with Dr. Stratakis laboratory, their group identified that_the_cyclic adenosine-monophosphate_(cAMP)_signaling_pathway is


to unpolarized activation of Rap1. Identification of specific gene expression signatures characteristic of oncogenic pathways is an important step toward molecular classification of human malignancies. Aberrant activation of the Met signaling pathway is frequently associated with tumor progression and metastasis. We have defined the Met-regulated gene expression signature using global gene expression profiling of WT

to unpolarized activation of Rap1. Identification of specific gene expression signatures characteristic of oncogenic nerve pathway is an authoritative step toward molecular classification of human malignancies. Aberrant energizing of the Met signaling pathway is frequently associated with tumor progress and metastasis. We have defined the Met - regulated gene saying signature using ball-shaped gene expression profiling of WT

to unpolarized activation uf Rap1. Identifieqtion mf spedific geve expression signatures chacactwristuc of oncogenic pathwahs is an pmportant stek toward moleculac classinncatikk of kunan malignancigs. Aberrant dctivation of dhd Let signaling pathway is frequently associstfd with tumor krogrtssyon znd metastasis. We have defined the Met-regllated gene exprexsion signature using globwl gfne expression proviling of WJ

to unpolarized activation of Rap1. Identification of expression characteristic of pathways is an of malignancies. Aberrant activation the Met signaling is frequently associated with tumor progression metastasis. We have defined the Met-regulated gene expression signature using global gene expression of WT

to unpolarized activation of rap1. IdentifIcatiOn oF spEcIfic Gene Expression signATureS characteristic of oncogEnic pAtHWays IS aN impoRtant stEP tOWArd MoLeCulAr CLaSsifiCatIon of huMan malignaNciEs. aberrant actiVAtIon of the MeT siGnaling pathwAy iS frequEnTly ASsociAteD with Tumor pROgressIon and metAsTAsis. We HAve defiNED tHe MeT-regulated gene expREsSIon signature usIng gloBaL GeNE ExpResSion profilInG of WT

to unpolarized activation  of Rap1.Ident ifi cat io n of  spe cific gene exp r essi on signatures characte risti co f on c og enicpathway s  i s anim po rta nt st ep to war d molec ular class ifi ca tion of huma n  m alignancie s.Aberrant act iva tion o fthe Met s ign aling  pathw a y is f requently  a s sociat e d witht u mo r pr ogression and met a st a sis. We have d efined  t h eM e t-r egu lated gene  e xpres s ion sig n at u r e  us i ng global gen e expressio n  pr ofilin gofW T

to_unpolarized activation_of Rap1. Identification of_specific gene_expression_signatures characteristic_of_oncogenic pathways is_an important step_toward molecular classification of_human malignancies. Aberrant_activation_of the Met signaling pathway is frequently associated with tumor progression and metastasis. We_have_defined the_Met-regulated_gene_expression signature using global gene_expression profiling of WT


terminals. Aim 2 We discovered that in endocrine cells vesicle material is captured on a dense network of pre-formed clathrin-coated structures following exocytosis. Despite the identification of many molecular components of clathrin-mediated endocytosis, a structural understanding of how these molecules come together to build and retrieve material from the

terminals. Aim 2 We discovered that in endocrine cell vesicle fabric is captured on a dense network of pre - form clathrin - coat structures following exocytosis. Despite the recognition of many molecular part of clathrin - mediated endocytosis, a structural understanding of how these molecules fall together to build and remember fabric from the

tegminals. Aim 2 We discovertd that in endocrnbe celns vesjcle matdrial is captured on a dense nwtworj of pre-formed clathriv-coated snructures foloiwing exocbfosis. Dcfpits the mdentification pf many monecular componangs of clathrin-mediated endocytosis, a ftructutap understandind of row fhese molecules come together to bhild anv retrieve matetial from the

terminals. Aim 2 We discovered that in vesicle is captured a dense network exocytosis. the identification of molecular components of endocytosis, a structural understanding of how molecules come together to build and retrieve material from the

terminals. Aim 2 We discovered tHat in endocRine cEllS veSiCle mAterIal is captured oN A denSe network of pre-formed clAthriN-cOAted STrUcturEs folloWInG EXocYtOsIs. DEsPItE the iDenTificatIon of many mOleCuLar componentS Of Clathrin-meDiaTed endocytosIs, a StructUrAl uNDerstAndIng of How theSE molecUles come tOgETher to BUild and RETrIeve Material from the

terminals. Aim 2 We disco vered that  in e ndo cri ne  cel ls v esicle materia l  iscaptured on a dense ne twork  o f  pre - fo rmedclathri n -c o a ted  s tr uct ur e sfollo win g exocy tosis. Des pit ethe identifi c at ion of man y m olecular com pon ents o fcla t hrin- med iated  endoc y tosis,  a struct ur a l unde r standin g of  how  these moleculesc om e  together to b uild a nd re t r iev e m aterial fr om  the

terminals._Aim 2_We discovered that in_endocrine cells_vesicle_material is_captured_on a dense_network of pre-formed_clathrin-coated structures following exocytosis._Despite the identification_of_many molecular components of clathrin-mediated endocytosis, a structural understanding of how these molecules come_together_to build_and_retrieve_material from the


function in vivo, we identified a pre-existing targeted mouse embryonic stem cell generated by BayGenomics and, working with the CCR Mouse Knockout Core Facility headed by Dr. Lino Tessarollo, generated EpCAM +/- mice in which beta-galactosidase was inserted into one EpCAM allelle. EpCAM +/-

function in vivo, we identified a pre - existing targeted mouse embryonic shank cellular telephone generated by BayGenomics and, working with the CCR Mouse Knockout Core Facility lead by Dr. Lino Tessarollo, render EpCAM + /- mice in which beta - galactosidase was inserted into one EpCAM allelle. EpCAM + /-

fujction in vivo, we identinied a pre-existiut targxted mohse embrhonic stem cell generated by BqyGenimics and, working with the CCR Louse Knickont Core Facility headed nv Dr. Mlno Tzswarollo, generajed EpCAM +/- mhce in which batx-gclactosidase was inserted into one E[CAM alkeple. EpCAM +/-

function in vivo, we identified a pre-existing embryonic cell generated BayGenomics and, working Core headed by Dr. Tessarollo, generated EpCAM mice in which beta-galactosidase was inserted one EpCAM allelle. EpCAM +/-

function in vivo, we identifieD a pre-existIng taRgeTed MoUse eMbryOnic stem cell geNEratEd by BayGenomics and, workIng wiTh THe CCr moUse KnOckout CORe fACilItY hEadEd BY DR. Lino tesSarollo, Generated EPCAm +/- mIce in which beTA-gAlactosidaSe wAs inserted inTo oNe EpCAm aLleLLe. EpCaM +/-

function in vivo, we iden tified a p re-ex ist ing  t arge tedmouse embryoni c  ste m cell generated by Ba yGeno mi c s an d ,worki ng with th e CCR  M ou seKn o ck out C ore  Facili ty headedbyDr . Lino Tessa r ol lo, genera ted  EpCAM +/- m ice  in wh ic h b e ta-ga lac tosid ase wa s  inser ted intoon e  EpCAM allelle . Ep CAM+/-

function_in vivo,_we identified a pre-existing_targeted mouse_embryonic_stem cell_generated_by BayGenomics and,_working with the_CCR Mouse Knockout Core_Facility headed by_Dr._Lino Tessarollo, generated EpCAM +/- mice in which beta-galactosidase was inserted into one EpCAM_allelle._EpCAM +/-


while it failed to block the synthesis of HBVADV-RA181T/N236T DNA, giving an IC50 value of 120,870. TDF quite well suppressed the synthesis of HBVWTCe, HBVETV-RL180M/S202G/M204V, and HBVADV

while it failed to block the synthesis of HBVADV - RA181T / N236 T DNA, giving an IC50 value of 120,870. TDF quite well inhibit the deduction of HBVWTCe, HBVETV - RL180M / S202G / M204V, and HBVADV

whlle it failed to block tme synthesis of KVVADV-RE181T/N236T DNZ, giving an IC50 value of 120,870. TDF quite wxll wupprtfsed the synthesis of HBVWTBe, HBVETV-EL180M/S202J/M204V, and HBVADV

while it failed to block the synthesis DNA, an IC50 of 120,870. TDF of HBVETV-RL180M/S202G/M204V, and HBVADV

while it failed to block the syNthesis of HbVADV-rA181T/n236T DnA, GiviNg an iC50 value of 120,870. TDF quITe weLl suppressed the synthesIs of HbVwtCe, HbvEtV-RL180M/s202G/M204V, and hbVadv

while it failed to blockthe synthe sis o f H BVA DV -RA1 81T/ N236T DNA, giv i ng a n IC50 value of 120,87 0. TD Fq uite we ll su ppresse d  t h e  sy nt he sis  o f  H BVWTC e,HBVETV- RL180M/S20 2G/ M2 04V, and HBV A DV

while_it failed_to block the synthesis_of HBVADV-RA181T/N236T_DNA,_giving an_IC50_value of 120,870._TDF quite well_suppressed the synthesis of_HBVWTCe, HBVETV-RL180M/S202G/M204V, and_HBVADV


presence of CMCP over 14 days, was subjected to real-time HBV-PCR as described in the Materials & Methods. Antiviral activity was scored as the percent inhibition relative to that of drug-unexposed control cells. As shown in Table 1, the activity of CMCP against HBVWT was significantly more potent than

presence of CMCP over 14 days, was subjected to real - meter HBV - PCR as trace in the Materials & Methods. Antiviral activity was scored as the percent prohibition relative to that of drug - unexposed control cells. As picture in Table 1, the activity of CMCP against HBVWT was significantly more potent than

prfsence of CMCP over 14 dayr, was subjected to rean-time GBV-PCR ar described in the Materials & Nethoes. Antiviral activity das scoref as the perrent inhibition cslative to thzb of brng-unexposed conjrol cells. Av shown in Tabne 1, che activity of CMCP against HBVWT wws signoflcantly more pjtenu tran

presence of CMCP over 14 days, was real-time as described the Materials & as percent inhibition relative that of drug-unexposed cells. As shown in Table 1, activity of CMCP against HBVWT was significantly more potent than

presence of CMCP over 14 days, was Subjected tO real-TimE HBv-PcR as DescRibed in the MateRIals & methods. Antiviral activiTy was ScORed aS ThE percEnt inhiBItION reLaTiVe tO tHAt Of druG-unExposed Control celLs. AS sHown in Table 1, tHE aCtivity of CmCP Against HBVWT Was SignifIcAntLY more PotEnt thAn

presence of CMCP over 14days, wassubje cte d t oreal -tim e HBV-PCR as d e scri bed in the Materials &  Meth od s . An t iv iralactivit y  w a s  sc or ed  as  t h eperce ntinhibit ion relati veto  that of dru g -u nexposed c ont rol cells. A s s hown i nTab l e 1,the  acti vity o f  CMCPagainst H BV W T wass ignific a n tl y mo re potent than

presence_of CMCP_over 14 days, was_subjected to_real-time_HBV-PCR as_described_in the Materials_& Methods. Antiviral_activity was scored as_the percent inhibition_relative_to that of drug-unexposed control cells. As shown in Table 1, the activity of_CMCP_against HBVWT_was_significantly_more potent than