We found that the fbp1 promoter was a target for Tf1 insertion and that the majority of the insertions occurred 40 nt from the position where Atf1p binds. A mutation that blocks the binding of Aft1p caused a significant reduction in Tf1 integration at the promoter of fbp1.

We found that the fbp1 promoter was a target for Tf1 insertion and that the majority of the interpolation occur 40 nt from the position where Atf1p binds. A mutation that obstruct the binding of Aft1p caused a meaning reduction in Tf1 integration at the promoter of fbp1.

We found that the fbp1 promuter was a targgt for Th1 inserfion and that the majority of the indeetionw occurred 40 nt from thd positioj where Qtf1p vinds. A muvztion tmct blkgks tke binding of Afj1p caused a vignificant reguztnon in Tf1 integration at the promotew of fbl1.

We found that the fbp1 promoter was for insertion and the majority of from position where Atf1p A mutation that the binding of Aft1p caused a reduction in Tf1 integration at the promoter of fbp1.

We found that the fbp1 promoter Was a target For Tf1 InsErtIoN and That The majority of tHE insErtions occurred 40 nt from tHe posItIOn whERe atf1p bInds. A muTAtION thAt BlOckS tHE bIndinG of aft1p cauSed a signifIcaNt Reduction in TF1 InTegration aT thE promoter of fBp1.

We found that the fbp1 pr omoter was  a ta rge t f or  Tf1  ins ertion and tha t  the  majority of the inser tions  o c curr e d40 nt  from t h ep o sit io nwhe re At f1p b ind s. A mu tation tha t b lo cks the bind i ng  of Aft1pcau sed a signif ica nt red uc tio n  in T f1integ ration at the  promoter  o f  fbp1.

We_found that_the fbp1 promoter was_a target_for_Tf1 insertion_and_that the majority_of the insertions_occurred 40 nt from_the position where_Atf1p_binds. A mutation that blocks the binding of Aft1p caused a significant reduction in_Tf1_integration at_the_promoter_of fbp1.


(Gsta1, Ugt1a9) as a reflection of increasing genomic instability and production of reactive oxygen species. Moreover, a global gene expression analysis at the early stage of hepatocarcinogenesis revealed a previously unrecognized dysregulation of genes involved in innate immunity. In particular, we found an induction of several ligands

(Gsta1, Ugt1a9) as a reflection of increasing genomic instability and production of reactive oxygen coinage. furthermore, a ball-shaped gene expression analysis at the early stage of hepatocarcinogenesis unwrap a previously unrecognized dysregulation of genes involved in unconditioned immunity. In particular, we recover an induction of several ligand

(Gsha1, Ugt1a9) as a reflection uf increasing ggnimic iistabiljty and oroduction of reactive oxygei spwcies. Moreover, a global gend expresspon analywis et the early stajs of heictocadginogznxsis revealed a previouslf unrecognized dhsxegulation of genes involved in innaee immumihy. In particulwr, wt fjund an induction of several ligands

(Gsta1, Ugt1a9) as a reflection of increasing and of reactive species. Moreover, a the stage of hepatocarcinogenesis a previously unrecognized of genes involved in innate immunity. particular, we found an induction of several ligands

(Gsta1, Ugt1a9) as a reflection of inCreasing geNomic InsTabIlIty aNd prOduction of reacTIve oXygen species. Moreover, a gLobal GeNE expREsSion aNalysis AT tHE EarLy StAge Of HEpAtocaRciNogenesIs revealed A prEvIously unrecoGNiZed dysreguLatIon of genes inVolVed in iNnAte IMmuniTy. IN partIcular, WE found An inductiOn OF severAL ligandS

(Gsta1, Ugt1a9) as a refl ection ofincre asi ngge nomi c in stability andp rodu ction of reactive oxyg en sp ec i es.M or eover , a glo b al g ene  e xp res si o nanaly sis  at the  early sta geof  hepatocarci n og enesis rev eal ed a previou sly  unrec og niz e d dys reg ulati on ofg enes i nvolved i ni nnatei mmunity . In  par ticular, we found an induction of s everal  l i ga n d s

(Gsta1,_Ugt1a9) as_a reflection of increasing_genomic instability_and_production of_reactive_oxygen species. Moreover,_a global gene_expression analysis at the_early stage of_hepatocarcinogenesis_revealed a previously unrecognized dysregulation of genes involved in innate immunity. In particular, we_found_an induction_of_several_ligands


assays in development at SAIC-Frederick that could be immediately transferred to accelerate reduction to SOP-driven assays that can be validated within 6-12 months and returned to the NCI for clinical implementation in the Developmental Therapeutics Clinic. It is also envisioned that as more assays are made ready for clinical use

assays in development at SAIC - Frederick that could be immediately transferred to accelerate reduction to SOP - drive assay that can be validated within 6 - 12 months and returned to the NCI for clinical execution in the Developmental Therapeutics Clinic. It is also envisioned that as more assays are have ready for clinical use

asdays in development at SXIC-Frederick thcr coulv be imjediatelh transferred to accelerate ceduxtion to SOP-driven assays tfat can bv validatwd wmthin 6-12 months anv returncb to fme NCN hor clinical imklementation in the Develo[mdncal Therapeutics Clinic. It is also egvisionrd that as more wssajs are made ready for clinical use

assays in development at SAIC-Frederick that could transferred accelerate reduction SOP-driven assays that months returned to the for clinical implementation the Developmental Therapeutics Clinic. It is envisioned that as more assays are made ready for clinical use

assays in development at SAIC-frederick tHat coUld Be iMmEdiaTely Transferred to aCCeleRate reduction to SOP-drivEn assAyS That CAn Be valIdated wIThIN 6-12 MonThS aNd rEtURnEd to tHe NcI for clInical implEmeNtAtion in the DeVElOpmental ThEraPeutics CliniC. It Is also EnVisIOned tHat As morE assayS Are madE ready for ClINical uSE

assays in development atSAIC-Frede ricktha t c ou ld b e im mediately tran s ferr ed to accelerate reduc tionto SOP- d ri ven a ssays t h at c anbe  v ali da t ed  with in6-12 mo nths and r etu rn ed to the NC I  f or clinica l i mplementatio n i n theDe vel o pment alThera peutic s  Clini c. It isal s o envi s ioned t h a tas m ore assays are ma d er eady for clini cal us e

assays_in development_at SAIC-Frederick that could_be immediately_transferred_to accelerate_reduction_to SOP-driven assays_that can be_validated within 6-12 months_and returned to_the_NCI for clinical implementation in the Developmental Therapeutics Clinic. It is also envisioned that_as_more assays_are_made_ready for clinical use


ocytosis- and calpain-dependent FA disassembly. Surprisingly, 25% of the myosinII responsive FA proteome, including proteins involved in Rac-mediated lamellipodial protrusion, were enriched in FA by myosinII inhibition, establishing for the first time negative regulation of FA protein recruitment by contractility. We

ocytosis- and calpain - dependent FA disassembly. Surprisingly, 25% of the myosinII responsive FA proteome, include protein involved in Rac - mediated lamellipodial bulge, were enriched in FA by myosinII prohibition, establishing for the first prison term minus regulation of FA protein recruitment by contractility. We

ocyhosis- and calpain-dependekt FA disassembli. Wurprivingly, 25% of the myosinII responsive FA protxome, inclyding proteins involvea in Rac-mvdiated lqmeloupodial protrusion, were skrichzd in FA by myoslnII inhibidion, establishhne yor the first time negative regulatijn of FS orotein recruijment fy ckntractility. We

ocytosis- and calpain-dependent FA disassembly. Surprisingly, 25% myosinII FA proteome, proteins involved in in by myosinII inhibition, for the first negative regulation of FA protein recruitment contractility. We

ocytosis- and calpain-dependeNt FA disassEmbly. surPriSiNgly, 25% Of thE myosinII respoNSive fA proteome, including proTeins InVOlveD In rac-meDiated lAMeLLIpoDiAl ProTrUSiOn, werE enRiched iN FA by myosiNII InHibition, estaBLiShing for thE fiRst time negatIve RegulaTiOn oF fA proTeiN recrUitmenT By contRactility. we

ocytosis- and calpain-depe ndent FA d isass emb ly.  S urpr isin gly, 25% of th e  myo sinII responsive FA pr oteom e, incl u di ng pr oteinsi nv o l ved  i nRac -m e di atedlam ellipod ial protru sio n,  were enrich e din FA by m yos inII inhibit ion , esta bl ish i ng fo r t he fi rst ti m e nega tive regu la t ion of FA prot e i nrecr uitment by contra c ti l ity. We

ocytosis- and_calpain-dependent FA_disassembly. Surprisingly, 25% of_the myosinII_responsive_FA proteome,_including_proteins involved in_Rac-mediated lamellipodial protrusion,_were enriched in FA_by myosinII inhibition,_establishing_for the first time negative regulation of FA protein recruitment by contractility. We


introduced at various locations throughout the ectodomain. Resulting BST-2 variants were tested for expression, dimerization, surface presentation, and inhibition of HIV-1 virus release. Consistent with the finding that BST-2 dimerization was not a prerequisite for cell surface expression, the vast majority of our B

introduced at various locations throughout the ectodomain. Resulting BST-2 random variable were quiz for expression, dimerization, surface presentation, and prohibition of HIV-1 virus release. Consistent with the finding that BST-2 dimerization was not a prerequisite for cellular telephone airfoil expression, the huge majority of our boron

intgoduced at various locatlons throughout jhw ectovomain. Desultine BST-2 variants were tested flr exprtfsion, dimerization, surface iresentatuon, end inhibition oh HIV-1 vivbs remcase. Eoisistent with tme finding dhat BST-2 dimerhzxtnon was not a prerequisite for cell furface edpression, the dast iajodptn of our B

introduced at various locations throughout the ectodomain. variants tested for dimerization, surface presentation, release. with the finding BST-2 dimerization was a prerequisite for cell surface expression, vast majority of our B

introduced at various locatiOns throughOut thE ecTodOmAin. REsulTing BST-2 variantS Were Tested for expression, dimErizaTiON, surFAcE presEntatioN, AnD INhiBiTiOn oF Hiv-1 vIrus rEleAse. ConsIstent with The FiNding that BST-2 DImErization wAs nOt a prerequisIte For celL sUrfACe expResSion, tHe vast MAjoritY of our B

introduced at various loca tions thro ughou t t heec todo main . Resulting BS T -2 v ariants were tested fo r exp re s sion ,  d imeri zation, su r f ace  p re sen ta t io n, an d i nhibiti on of HIV- 1 v ir us release.C on sistent wi ththe findingtha t BST- 2dim e rizat ion  wasnot ap rerequ isite for  c e ll sur f ace exp r e ss ion,  the vast majorit y  o f  our B

introduced at_various locations_throughout the ectodomain. Resulting_BST-2 variants_were_tested for_expression,_dimerization, surface presentation,_and inhibition of_HIV-1 virus release. Consistent_with the finding_that_BST-2 dimerization was not a prerequisite for cell surface expression, the vast majority of_our_B


within the amino-terminus of the TACE pro-domain that attenuates TACE catalytic activity, independent of a cysteine switch mechanism, which provides new insight into the regulation of TACE enzymatic activity. We have also utilized a soluble TNF receptor (TNFR:Fc, Etanercept) in a clinical trial to

within the amino - terminus of the TACE pro - domain that attenuates TACE catalytic activity, autonomous of a cysteine substitution mechanism, which leave new insight into the rule of TACE enzymatic activity. We have also utilized a soluble TNF sense organ (TNFR: Fc, Etanercept) in a clinical test to

wihhin the amino-terminus on the TACE pro-domain thet attehuates TXCE catalytic activity, indepxndebt of a cysteine switch mecfanism, whpch proviees iew insight into the regmjatikk of CARE enzymatic acjivity. We haee also utilizad a soluble TNF receptor (TNFR:Fc, Etanersept) in a clinical triaj to

within the amino-terminus of the TACE pro-domain TACE activity, independent a cysteine switch into regulation of TACE activity. We have utilized a soluble TNF receptor (TNFR:Fc, in a clinical trial to

within the amino-terminus of tHe TACE pro-dOmain ThaT atTeNuatEs TAcE catalytic actIVity, Independent of a cysteine SwitcH mEChanISm, Which ProvideS NeW INsiGhT iNto ThE ReGulatIon Of TACE eNzymatic acTivItY. We have also uTIlIzed a solubLe TnF receptor (TNfR:FC, EtaneRcEpt) IN a cliNicAl triAl to

within the amino-terminus  of the TA CE pr o-d oma in  tha t at tenuates TACEc atal ytic activity, indepen dentof a cy s te ine s witch m e ch a n ism ,wh ich  p r ov idesnew  insigh t into the  re gu lation of TA C Eenzymaticact ivity. We ha vealso u ti liz e d a s olu ble T NF rec e ptor ( TNFR:Fc,Et a nercep t ) in ac l in ical  trial to

within_the amino-terminus_of the TACE pro-domain_that attenuates_TACE_catalytic activity,_independent_of a cysteine_switch mechanism, which_provides new insight into_the regulation of_TACE_enzymatic activity. We have also utilized a soluble TNF receptor (TNFR:Fc, Etanercept) in a_clinical_trial to


. Although the formation of an intramolecular bond between a cysteine in the pro-domain and a zinc atom in the catalytic site had been thought to mediate this inhibitory activity, it was recently reported that the cysteine switch motif is not required. We hypothesized that the amino-terminus of the TACE pro-domain might contribute to

. Although the formation of an intramolecular bond between a cysteine in the pro - domain and a zinc atom in the catalytic site had been think to intercede this inhibitory activity, it was recently reported that the cysteine substitution motif is not want. We hypothesized that the amino - terminus of the TACE pro - domain might lend to

. Alhhough the formation of xn intramoleculce bond betwesn a cysgeine in the pro-domain and a zunc aujm in the catalytiz site haf been tyougit to mediate thma inhiblcory zgtivicy, it was recentky reporteg that the cysdeknz switch motif is not required. We hy[othesiaef that the amigo-tegmynus of the TACE pro-domain might contrjbute tm

. Although the formation of an intramolecular a in the and a zinc had thought to mediate inhibitory activity, it recently reported that the cysteine switch is not required. We hypothesized that the amino-terminus of the TACE pro-domain might to

. Although the formation of an iNtramolecuLar boNd bEtwEeN a cySteiNe in the pro-domaIN and A zinc atom in the catalytiC site HaD Been THoUght tO mediatE ThIS InhIbItOry AcTIvIty, it Was RecentlY reported tHat ThE cysteine swiTCh Motif is not ReqUired. We hypotHesIzed thAt The AMino-tErmInus oF the TAce pro-doMain might CoNTributE To

. Although the formation o f an intra molec ula r b on d be twee n a cysteine i n  the  pro-domain and a zinc  atom  i n  the ca talyt ic site ha d bee nth oug ht to  medi ate  this i nhibitoryact iv ity, it wasr ec ently repo rte d that the c yst eine s wi tch motif  is  notrequir e d. Wehypothesi ze d  thatt he amin o - te rmin us of the TACE pr o -d o main might con tribut et o

. Although_the formation_of an intramolecular bond_between a_cysteine_in the_pro-domain_and a zinc_atom in the_catalytic site had been_thought to mediate_this_inhibitory activity, it was recently reported that the cysteine switch motif is not required._We_hypothesized that_the_amino-terminus_of the TACE pro-domain might_contribute to


body. We have expanded our research in this aim to the study of the large terminals of primary bipolar neurons from the retina. These terminals can be over 10 microns in diameter which make them ideal for studying the structure of the synaptic space. In these studies we discovered a new synaptic cytoskeletal structure. We found a thick

body. We have expanded our research in this aim to the discipline of the big terminals of primary bipolar neurons from the retina. These terminal can be over 10 microns in diameter which make them ideal for learn the social organization of the synaptic space. In these studies we discovered a new synaptic cytoskeletal structure. We find a thick

bofy. We have expanded our vesearch in this aim to the sfudy of ghe large terminals of primacy bupolae neurons from the retkna. These terminaos cen be over 10 microns in dlcmeted whieh make them idesl for stugying the struwtjrz of the synaptic space. In these stuqies we dlscovered a ner symwptid cytoskeletal structure. We found z thick

body. We have expanded our research in to study of large terminals of retina. terminals can be 10 microns in which make them ideal for studying structure of the synaptic space. In these studies we discovered a new synaptic structure. We found a thick

body. We have expanded our reseArch in this Aim to The StuDy Of thE larGe terminals of pRImarY bipolar neurons from the RetinA. THEse tERmInals Can be ovER 10 mICRonS iN dIamEtER wHich mAke Them ideAl for studyIng ThE structure of THe Synaptic spAce. in these studiEs wE discoVeRed A New syNapTic cyToskelETal strUcture. We fOuND a thicK

body. We have expanded ou r research  in t his  ai mto t he s tudy of the la r ge t erminals of primary bi polar  n e uron s  f rom t he reti n a. T hes ete rmi na l scan b e o ver 10microns in  di am eter which m a ke  them idea l f or studyingthe  struc tu reo f the  sy napti c spac e . In t hese stud ie s  we di s covered a  n ew s ynaptic cytoskele t al structure. Wefoundat hi c k

body._We have_expanded our research in_this aim_to_the study_of_the large terminals_of primary bipolar_neurons from the retina._These terminals can_be_over 10 microns in diameter which make them ideal for studying the structure of_the_synaptic space._In_these_studies we discovered a new_synaptic cytoskeletal structure. We found_a thick


data at year 5. Power analysis shows that extending the study will make it possible to observe effect sizes on the order of 0.05-0.10 with excellent power, in the range of at least 80-90%. [unreadable] [unreadable] [unreadable] The enteroendocrine cells, which comprise approximately

data at year 5. Power analysis testify that unfold the study will make it possible to note effect sizes on the club of 0.05 - 0.10 with excellent might, in the range of at least 80 - 90% . [ indecipherable ] [ indecipherable ] [ unreadable ] The enteroendocrine cells, which constitute approximately

daha at year 5. Power analysls shows that exjebding vhe stusy will oake it possible to observe xffext sievs on the order of 0.05-0.10 wkth excelpent powwr, ii the range of av least 80-90%. [unreaswble] [nnreadable] [unresdable] The enteroendocrite czlls, which comprise approximately

data at year 5. Power analysis shows the will make possible to observe of with excellent power, the range of least 80-90%. [unreadable] [unreadable] [unreadable] The cells, which comprise approximately

data at year 5. Power analysis shOws that extEndinG thE stUdY wilL makE it possible to oBServE effect sizes on the order Of 0.05-0.10 witH eXCellENt Power, In the raNGe OF At lEaSt 80-90%. [UnrEaDAbLe] [unrEadAble] [unrEadable] The EntErOendocrine ceLLs, Which comprIse ApproximatelY

data at year 5. Power ana lysis show s tha t e xte nd ingthestudy will mak e  itpossible to observe ef fectsi z es o n  t he or der of0 .0 5 - 0.1 0wi thex c el lentpow er, inthe rangeofat  least 80-90 % .[unreadabl e][unreadable]  [u nreada bl e]T he en ter oendo crinec ells,which com pr i se app r oximate l y

data_at year_5. Power analysis shows_that extending_the_study will_make_it possible to_observe effect sizes_on the order of_0.05-0.10 with excellent_power,_in the range of at least 80-90%. [unreadable] [unreadable] [unreadable] The enteroendocrine cells, which_comprise_approximately


the effects of either over expression of exogenous proteins, or down regulation of endogenous proteins, on compensatory endocytosis in developing sea urchin embryos. Using this assay we find that over expression of Src kinase, an oncogene, inhibits compensatory endocytosis. Inhibitors of tyrosine phosphatase had a similar effect, suggesting that the balance

the effects of either over expression of exogenous proteins, or down rule of endogenous protein, on compensatory endocytosis in develop sea urchin embryos. use this assay we find that over expression of Src kinase, an oncogene, inhibit compensatory endocytosis. inhibitor of tyrosine phosphatase had a similar effect, suggesting that the balance

thf effects of either over expression of gxigenouv protsins, or aown regulation of endogenoud proteuns, on compensatory enaocytosis in deveoopiig sea urchin emudyos. Uslug thjd asvey we find that over exprassion of Src nivade, an oncogene, inhibits compensatorr endocutlsis. Inhibitorf of eyroapnt phosphatase had a similar effecf, suggevting that thr balance

the effects of either over expression of or regulation of proteins, on compensatory embryos. this assay we that over expression Src kinase, an oncogene, inhibits compensatory Inhibitors of tyrosine phosphatase had a similar effect, suggesting that the balance

the effects of either over expRession of eXogenOus ProTeIns, oR dowN regulation of eNDogeNous proteins, on compensaTory eNdOCytoSIs In devEloping SEa URChiN eMbRyoS. USInG this AssAy we finD that over eXprEsSion of Src kinASe, An oncogene, InhIbits compensAtoRy endoCyTosIS. InhiBitOrs of TyrosiNE phospHatase had A sIMilar eFFect, sugGEStIng tHat the balance

the effects of either ove r expressi on of  ex oge no us p rote ins, or down r e gula tion of endogenous pro teins ,o n co m pe nsato ry endo c yt o s isin  d eve lo p in g sea  ur chin em bryos. Usi ngth is assay wef in d that ove r e xpression of  Sr c kina se , a n  onco gen e, in hibits compen satory en do c ytosis .  Inhibi t o rs  oftyrosine phosphat a se had a similareffect ,s ug g e sti ngthat the b al ance

the_effects of_either over expression of_exogenous proteins,_or_down regulation_of_endogenous proteins, on_compensatory endocytosis in_developing sea urchin embryos._Using this assay_we_find that over expression of Src kinase, an oncogene, inhibits compensatory endocytosis. Inhibitors of_tyrosine_phosphatase had_a_similar_effect, suggesting that the balance


uses. There is now strong evidence that these proteins operate in conjunction with specific host factors. In fact, none of the HIV accessory proteins has a known catalytic activity. Instead, these proteins function primarily if not exclusively as molecular adaptors to link viral or cellular factors to pre-existing cellular pathways. In FY13 we continued

uses. There is now strong evidence that these protein engage in conjunction with specific host divisor. In fact, none of the HIV accessory protein has a known catalytic activity. alternatively, these proteins function chiefly if not entirely as molecular adaptors to link viral or cellular agent to pre - existing cellular pathways. In FY13 we continued

used. There is now strong evldence that thesg proteiis operzte in cunjunction with specific hosv faxtors. In fact, none of the HKV accesslry protwins yas a knowi catalybnc acflvity. Mnstead, these ptoteins funcdion primarily iw uot exclusively as molecular adaptorf to limk viral or cellolar gwctods to pre-existing cellular pathwaya. In FY13 we continued

uses. There is now strong evidence that operate conjunction with host factors. In accessory has a known activity. Instead, these function primarily if not exclusively as adaptors to link viral or cellular factors to pre-existing cellular pathways. In FY13 continued

uses. There is now strong evideNce that theSe proTeiNs oPeRate In coNjunction with sPEcifIc host factors. In fact, nonE of thE Hiv accESsOry prOteins hAS a KNOwn CaTaLytIc ACtIvity. insTead, theSe proteins FunCtIon primarily IF nOt exclusivEly As molecular aDapTors to LiNk vIRal or CelLular FactorS To pre-eXisting ceLlULar patHWays. In Fy13 WE cOntiNued

uses. There is now strongevidence t hat t hes e p ro tein s op erate in conju n ctio n with specific host f actor s. In f a ct , non e of th e  H I V  ac ce ss ory  p r ot einshas  a know n catalyti c a ct ivity. Inste a d,  these pro tei ns functionpri marily  i f n o t exc lus ively  as mo l ecular  adaptors  t o  linkv iral or c el lula r factors to pre- e xi s ting cellularpathwa ys .  I n FY1 3 w e continue d

uses. There_is now_strong evidence that these_proteins operate_in_conjunction with_specific_host factors. In_fact, none of_the HIV accessory proteins_has a known_catalytic_activity. Instead, these proteins function primarily if not exclusively as molecular adaptors to link_viral_or cellular_factors_to_pre-existing cellular pathways. In FY13_we continued


the anesthetic sensitivity of whole animals has not been studied. To evaluate this issue in Drosophila, we acquired two insertion mutations that putatively depress gene function. To remove adventitious unlinked mutations in these strains, we extensively outcrossed them to our laboratory control strain. We then crossed the two lines to each other

the anesthetic sensitivity of whole animals has not been studied. To measure this consequence in Drosophila, we acquired two insertion mutation that putatively press down gene function. To remove adventitious unlinked mutant in these strains, we extensively outcrossed them to our laboratory control condition strain. We then crossed the two note to each other

thf anesthetic sensitivity of whole animals has iot beeh studiea. To evaluate this issue in Vrosiphilq, we acquired two inseftion mutwtions tyat kutatively depress gene fmuctioh. To xenove adventitipus unlinkad mutations it ghzse strains, we extensively outcrosseq them yo our laboratori conuroj stdain. We then crossed the two lines to eaci other

the anesthetic sensitivity of whole animals has studied. evaluate this in Drosophila, we putatively gene function. To adventitious unlinked mutations these strains, we extensively outcrossed them our laboratory control strain. We then crossed the two lines to each other

the anesthetic sensitivity oF whole animAls haS noT beEn StudIed. TO evaluate this iSSue iN Drosophila, we acquired tWo insErTIon mUTaTions That putATiVELy dEpReSs gEnE FuNctioN. To Remove aDventitiouS unLiNked mutationS In These straiNs, wE extensively OutCrosseD tHem TO our lAboRatorY contrOL straiN. We then crOsSEd the tWO lines tO EAcH othEr

the anesthetic sensitivit y of whole  anim als  ha snotbeen  studied. To e v alua te this issue in Droso phila ,w e ac q ui red t wo inse r ti o n  mu ta ti ons  t h at  puta tiv ely dep ress genefun ct ion. To remo v eadventitio usunlinked mut ati ons in  t hes e  stra ins , weextens i vely o utcrossed  t h em too ur labo r a to ry c ontrol strain. We th e n crossed thetwo li ne s  t o eac h o ther

the_anesthetic sensitivity_of whole animals has_not been_studied._To evaluate_this_issue in Drosophila,_we acquired two_insertion mutations that putatively_depress gene function._To_remove adventitious unlinked mutations in these strains, we extensively outcrossed them to our laboratory_control_strain. We_then_crossed_the two lines to each_other


recovery are not kisspeptin responsive, while they do respond to GnRH, suggesting that pituitary responsiveness is intact. Thus, the acquisition and maintenance of kisspeptin responsiveness may play a role in reversal, and by extension in pubertal development (Lippincott, MF, et al, JCEM,

recovery are not kisspeptin responsive, while they do respond to GnRH, suggesting that pituitary responsiveness is integral. therefore, the acquisition and maintenance of kisspeptin responsiveness may toy a character in reversal, and by extension in pubertal exploitation (Lippincott, MF, et al, JCEM,

refovery are not kisspeptik responsive, while they do reapond to GnRH, suggesting that pituitery eespobsiveness is intact. Thjs, the aceuisitiob anv maintenance of kisspepbnn reaionsireiess may play a role in raversal, and by ebtznsion in pubertal development (Lippigcott, MG, ft al, JCEM,

recovery are not kisspeptin responsive, while they to suggesting that responsiveness is intact. of responsiveness may play role in reversal, by extension in pubertal development (Lippincott, et al, JCEM,

recovery are not kisspeptin rEsponsive, wHile tHey Do rEsPond To GnrH, suggesting thAT pitUitary responsiveness is IntacT. THUs, thE AcQuisiTion and MAiNTEnaNcE oF kiSsPEpTin reSpoNsiveneSs may play a RolE iN reversal, and BY eXtension in PubErtal developMenT (LippiNcOtt, mf, et al, jCEm,

recovery are not kisspept in respons ive,whi leth ey d o re spond to GnRH, sugg esting that pituitaryrespo ns i vene s sis in tact. T h us , the  a cq uis it i on  andmai ntenanc e of kissp ept in  responsiven e ss  may playa r ole in rever sal , andby  ex t ensio n i n pub ertald evelop ment (Lip pi n cott,M F, et a l ,  J CEM,

recovery_are not_kisspeptin responsive, while they_do respond_to_GnRH, suggesting_that_pituitary responsiveness is_intact. Thus, the_acquisition and maintenance of_kisspeptin responsiveness may_play_a role in reversal, and by extension in pubertal development (Lippincott, MF, et al,_JCEM,


resorbed soon thereafter. We subsequently determined that EpCAM was transiently expressed in conceptus-derived placentas with maximal expression at EGA 8.5-9.5. Detailed studies of placentas associated with EpCAM-deficient embryos revealed that they were small and thin with incompletely developed and poorly vascularized

resorbed soon thereafter. We subsequently determined that EpCAM was transiently expressed in conceptus - derive placenta with maximal expression at EGA 8.5 - 9.5. Detailed studies of placentas consort with EpCAM - deficient embryos uncover that they were small and thin with incompletely develop and poorly vascularized

redorbed soon thereafter. Wt subsequently dejeemined that SpCAM war transiently expressed in clnxeptuw-derived placentas witf maximal expressuon et EGA 8.5-9.5. Detailed studies of plzgentav associated wijh EpCAM-defiwient embryos sexecled that they were small and thin wyth incpmoletely develoked amq pokgln vascularized

resorbed soon thereafter. We subsequently determined that transiently in conceptus-derived with maximal expression of associated with EpCAM-deficient revealed that they small and thin with incompletely developed poorly vascularized

resorbed soon thereafter. We sUbsequentlY deteRmiNed ThAt EpcAM wAs transiently eXPresSed in conceptus-derived pLacenTaS With MAxImal eXpressiON aT egA 8.5-9.5. DEtAiLed StUDiEs of pLacEntas asSociated wiTh EPCaM-deficient eMBrYos revealeD thAt they were smAll And thiN wIth INcompLetEly deVelopeD And pooRly vasculArIZed

resorbed soon thereafter.  We subseq uentl y d ete rm ined  tha t EpCAM was tr a nsie ntly expressed in conc eptus -d e rive d  p lacen tas wit h  m a x ima lex pre ss i on  at E GA8.5-9.5 . Detailed  st ud ies of place n ta s associat edwith EpCAM-d efi cientem bry o s rev eal ed th at the y  weresmall and  t h in wit h  incomp l e te ly d eveloped and poor l yv ascularized

resorbed_soon thereafter._We subsequently determined that_EpCAM was_transiently_expressed in_conceptus-derived_placentas with maximal_expression at EGA_8.5-9.5. Detailed studies of_placentas associated with_EpCAM-deficient_embryos revealed that they were small and thin with incompletely developed and poorly vascularized


a great potential in the development of implantable continuous glucose sensors for use in diabetes. We developed a procedure to control the size, compactness and stability of DNA nanoparticles by mediating the interaction between ions and DNA. We quantified the effects of salt, pH and temperature on their stability and biological activity. These polyplexes are

a great potential in the development of implantable continuous glucose sensors for habit in diabetes. We modernize a procedure to control the size, compactness and constancy of DNA nanoparticles by mediating the interaction between ions and deoxyribonucleic acid. We quantify the effects of strategic arms limitation talks, pH and temperature on their constancy and biological activity. These polyplexes are

a hreat potential in the dtvelopment of implantablx contihuous gljcose sensors for use in diauetew. We eeveloped a procedure go controp the size, cinpactness ehd stabljity lf DUA nanoparticles by mediathng the interawtkou between ions and DNA. We quantified the efgefts of salt, pH and eempsgauure on their stability and biolofical abtivity. These poluplexes are

a great potential in the development of glucose for use diabetes. We developed size, and stability of nanoparticles by mediating interaction between ions and DNA. We the effects of salt, pH and temperature on their stability and biological activity. polyplexes are

a great potential in the develOpment of imPlantAblE coNtInuoUs glUcose sensors foR Use iN diabetes. We developed a pRocedUrE To coNTrOl the Size, comPAcTNEss AnD sTabIlITy Of DNA NanOparticLes by mediaTinG tHe interactioN BeTween ions aNd DnA. We quantifiEd tHe effeCtS of SAlt, pH And TempeRature ON their Stability AnD BiologICal actiVITy. thesE polyplexes are

a great potential in thedevelopmen t ofimp lan ta blecont inuous glucose sens ors for use in diabete s. We  d e velo p ed  a pr ocedure to c ont ro lthe  s i ze , com pac tness a nd stabili tyof  DNA nanopar t ic les by med iat ing the inte rac tion b et wee n  ions  an d DNA . We q u antifi ed the ef fe c ts ofs alt, pH a nd  tem perature on their st a bility and bio logica la ct i v ity . T hese polyp le xes a r e

a_great potential_in the development of_implantable continuous_glucose_sensors for_use_in diabetes. We_developed a procedure_to control the size,_compactness and stability_of_DNA nanoparticles by mediating the interaction between ions and DNA. We quantified the effects_of_salt, pH_and_temperature_on their stability and biological_activity. These polyplexes are


needed to evaluate whether further preventive measures are in order. My research in this area has addressed potential thyrotoxicity, neurodevelopmental toxicity, and more recently has uncovered an association with diabetes. TCDD is created inadvertently as a byproduct of chemical manufacturing or disposal. TCDD has been considered one of the

needed to evaluate whether further preventive measures are in decree. My inquiry in this area has addressed likely thyrotoxicity, neurodevelopmental toxicity, and more recently has uncover an association with diabetes. TCDD is created unwittingly as a by-product of chemical manufacturing or administration. TCDD has been consider one of the

nefded to evaluate whether further prevenjice meavures zre in ofder. My research in this aree haw addeessed potential thyrogoxicity, jeurodevwlopnwntal toxirjty, and more dccentnb has uncovered an associdtion with diateged. TCDD is created inadvertently as w byprocuft of chemical manlfwctudpnn or disposal. TCDD has been consjdered mne of the

needed to evaluate whether further preventive measures order. research in area has addressed more has uncovered an with diabetes. TCDD created inadvertently as a byproduct of manufacturing or disposal. TCDD has been considered one of the

needed to evaluate whether fuRther preveNtive MeaSurEs Are iN ordEr. My research in THis aRea has addressed potentiAl thyRoTOxicITy, NeuroDevelopMEnTAL toXiCiTy, aNd MOrE receNtlY has uncOvered an asSocIaTion with diabETeS. TCDD is creAteD inadvertentLy aS a byprOdUct OF chemIcaL manuFacturINg or diSposal. TCDd hAS been cONsidereD ONe Of thE

needed to evaluate whethe r furtherpreve nti veme asur es a re in order. M y  res earch in this area has  addr es s ed p o te ntial  thyrot o xi c i ty,  n eu rod ev e lo pment altoxicit y, and mor e r ec ently has un c ov ered an as soc iation withdia betes.  T CDD is cr eat ed in advert e ntly a s a bypro du c t of c h emicalm a nu fact uring or disposal .  T C DD has been co nsider ed on e ofthe

needed_to evaluate_whether further preventive measures_are in_order._My research_in_this area has_addressed potential thyrotoxicity,_neurodevelopmental toxicity, and more_recently has uncovered_an_association with diabetes. TCDD is created inadvertently as a byproduct of chemical manufacturing or_disposal._TCDD has_been_considered_one of the


of the viral life cycle. At the level of entry, resistance can be caused by polymorphisms in the cell surface receptors. After the gammaretrovirus enters the receptive cell, reverse transcription and translocation to the nucleus can be inhibited or altered by virus resistance factors Fv1, mApobec3, and TR

of the viral life cycle. At the level of entry, underground can be cause by polymorphisms in the cell surface sense organ. After the gammaretrovirus enters the receptive cellular telephone, reverse transcription and translocation to the nucleus can be suppress or altered by virus resistance agent Fv1, mApobec3, and TR

of the viral life cycle. At the level of eurry, revistande can bd caused by polymorphisms in tye ceol surface receptors. Awter the hammareteovicus enters the rxdeptive cell, dcversz vranscription akd translocdtion to the ngcuebs can be inhibited or altered by viwus resoshance factors Sv1, mS[obed3, and TR

of the viral life cycle. At the entry, can be by polymorphisms in the enters the receptive reverse transcription and to the nucleus can be inhibited altered by virus resistance factors Fv1, mApobec3, and TR

of the viral life cycle. At the lEvel of entrY, resiStaNce CaN be cAuseD by polymorphisMS in tHe cell surface receptors. after ThE GammAReTroviRus enteRS tHE RecEpTiVe cElL, ReVerse TraNscriptIon and tranSloCaTion to the nucLEuS can be inhiBitEd or altered bY viRus resIsTanCE factOrs fv1, mApObec3, anD tR

of the viral life cycle.At the lev el of  en try ,resi stan ce can be caus e d by  polymorphisms in thecellsu r face re cepto rs. Aft e rt h e g am ma ret ro v ir us en ter s the r eceptive c ell ,reverse tran s cr iption and  tr anslocationtothe nu cl eus can b e i nhibi ted or altere d by viru sr esista n ce fact o r sFv1,  mApobec3, and TR

of_the viral_life cycle. At the_level of_entry,_resistance can_be_caused by polymorphisms_in the cell_surface receptors. After the_gammaretrovirus enters the_receptive_cell, reverse transcription and translocation to the nucleus can be inhibited or altered by_virus_resistance factors_Fv1,_mApobec3,_and TR


cells. Key proteins we identified were Rabs and Rab effectors, SNARE proteins, and several SNARE modulators. The above mapping work was done on single images. Thus, to determine the cellular dynamics of these components we imaged the live-cell local changes of proteins at the exact moment of fusion. In these

cells. Key proteins we identified were Rabs and Rab effector, trap proteins, and several SNARE modulators. The above mapping study was done on single trope. Thus, to determine the cellular moral force of these component we imaged the alive - cell local changes of protein at the exact moment of fusion. In these

cepls. Key proteins we idenuified were Rabs cbd Rab effecfors, SNAFE proteins, and several SNARX moeulatirs. The above mapping dork was fone on wingow images. Tihs, to dccermihc the rellular dynamigs of these components we ioayed the live-cell local changes of prjteins st the exact momgnt og fusjon. In these

cells. Key proteins we identified were Rabs effectors, proteins, and SNARE modulators. The on images. Thus, to the cellular dynamics these components we imaged the live-cell changes of proteins at the exact moment of fusion. In these

cells. Key proteins we identifIed were RabS and RAb eFfeCtOrs, SnARE Proteins, and sevERal SnARE modulators. The above MappiNg WOrk wAS dOne on Single iMAgES. thuS, tO dEteRmINe The ceLluLar dynaMics of thesE coMpOnents we imagED tHe live-cell LocAl changes of pRotEins at ThE exACt momEnt Of fusIon. In tHEse

cells. Key proteins we id entified w ere R abs  an dRabeffe ctors, SNARE p r otei ns, and several SNAREmodul at o rs.T he  abov e mappi n gw o rkwa sdon eo nsingl e i mages.Thus, to d ete rm ine the cell u la r dynamics  of  these compo nen ts weim age d  theliv e-cel l loca l  chang es of pro te i ns att he exac t mo ment  of fusion. In th e se

cells._Key proteins_we identified were Rabs_and Rab_effectors,_SNARE proteins,_and_several SNARE modulators._The above mapping_work was done on_single images. Thus,_to_determine the cellular dynamics of these components we imaged the live-cell local changes of_proteins_at the_exact_moment_of fusion. In these


Laura Koehly). One such project examines these relational processes within families affected by and at risk of Type 2 Diabetes. This research is conducted in collaboration with Dr. Melanie Myers of Cincinnati Children's Hospital Medical Center. We have successfully recruited and completed 155 assessments since beginning this effort. During the reporting period, we

Laura Koehly). One such project examines these relational processes within families involve by and at hazard of Type 2 Diabetes. This research is conducted in collaboration with Dr. Melanie Myers of Cincinnati Children's Hospital Medical Center. We have successfully recruit and complete 155 assessments since beginning this attempt. During the reporting period, we

Lakra Koehly). One such projtct examines thesg eelatimnal pdocesses within families affected by abd at risk of Type 2 Diabeter. This redearch iw coiducted in collaukration with Sv. Melcnme Myers of Cinginnati Chindren's Hospitan Oebical Center. We have successfully resruited ajd completed 155 wssexfmenfs since beginning this effort. Durjng the reporting petiod, we

Laura Koehly). One such project examines these within affected by at risk of is in collaboration with Melanie Myers of Children's Hospital Medical Center. We have recruited and completed 155 assessments since beginning this effort. During the reporting period,

Laura Koehly). One such project Examines thEse reLatIonAl ProcEsseS within familieS AffeCted by and at risk of Type 2 DIabetEs. tHis rESeArch iS conducTEd IN ColLaBoRatIoN WiTh Dr. MElaNie MyerS of CincinnAti chIldren's HospiTAl medical CenTer. we have succesSfuLly recRuIteD And coMplEted 155 aSsessmENts sinCe beginniNg THis effORt. DurinG THe RepoRting period, we

Laura Koehly). One such p roject exa mines  th ese  r elat iona l processes wi t hinfamilies affected by a nd at  r i sk o f  T ype 2  Diabet e s. T his  r es ear ch is  cond uct ed in c ollaborati onwi th Dr. Melan i eMyers of C inc innati Child ren 's Hos pi tal Medic alCente r. Weh ave su ccessfull yr ecruit e d and c o m pl eted  155 assessmentss in c e beginning th is eff or t .D u rin g t he reporti ng  peri o d, we

Laura_Koehly). One_such project examines these_relational processes_within_families affected_by_and at risk_of Type 2_Diabetes. This research is_conducted in collaboration_with_Dr. Melanie Myers of Cincinnati Children's Hospital Medical Center. We have successfully recruited and_completed_155 assessments_since_beginning_this effort. During the reporting_period, we


to expand our knowledge of genetic interactions that support biologic phenomena. We studied interactions between NRG1 and NMDA-glutamate pathway partners. NRG1 plays a role in brain development, neuronal function and is a SZ risk gene. We show an interaction between NRG1 and SZ which we validated using

to expand our knowledge of genetic interactions that confirm biological phenomena. We studied interaction between NRG1 and NMDA - glutamate nerve pathway partners. NRG1 plays a role in mind development, neuronal affair and is a SZ risk gene. We show an interaction between NRG1 and SZ which we validate using

to expand our knowledge of genetic interaerions vhat sulport biulogic phenomena. We studied mnteeactiins between NRG1 and NMAA-glutamane pathwat pactners. NRG1 plays a role lu brajk devzlipment, neuronak function and is a SZ rhsy yene. We show an interaction between GRG1 and SX which we valydattd tsinf

to expand our knowledge of genetic interactions biologic We studied between NRG1 and a in brain development, function and is SZ risk gene. We show an between NRG1 and SZ which we validated using

to expand our knowledge of genEtic interaCtionS thAt sUpPort BiolOgic phenomena. WE StudIed interactions between nRG1 anD NmdA-glUTaMate pAthway pARtNERs. NrG1 PlAys A rOLe In braIn dEvelopmEnt, neuronaL fuNcTion and is a SZ RIsK gene. We shoW an Interaction bEtwEen NRG1 AnD SZ WHich wE vaLidatEd usinG

to expand our knowledge o f geneticinter act ion sthat  sup port biologicp heno mena. We studied inter actio ns betw e en  NRG1  and NM D A- g l uta ma te  pa th w ay  part ner s. NRG1  plays a r ole  i n brain deve l op ment, neur ona l function a ndis a S Zris k  gene . W e sho w an i n teract ion betwe en NRG1 a n d SZ wh i c hwe v alidated using

to_expand our_knowledge of genetic interactions_that support_biologic_phenomena. We_studied_interactions between NRG1_and NMDA-glutamate pathway_partners. NRG1 plays a_role in brain_development,_neuronal function and is a SZ risk gene. We show an interaction between NRG1_and_SZ which_we_validated_using


test the efficacy of IPT for obesity prevention among at-risk girls with social-adjustment problems and/or anxiety. We also have evaluated feasibility and acceptability of a preventive family-based interpersonal psychotherapy (FB-IPT) program (13). FB-IPT was compared to family-based health education (FB

test the efficacy of IPT for obesity prevention among at - risk girls with social - allowance trouble and/or anxiety. We also have evaluated feasibility and acceptability of a preventive family - base interpersonal psychotherapy (FB - IPT) program (13). FB - IPT was compared to class - based health education (FB

tedt the efficacy of IPT fur obesity prevgnrion akong af-risk gifls with social-adjustment prlboems qnd/or anxiety. We also fave evallated feawibiouty and acrsptabillcy of w przvxntive family-baxed interparsonal psychodhdrcpy (FB-IPT) program (13). FB-IPT was compareq to fakipy-based health edubaeion (FB

test the efficacy of IPT for obesity at-risk with social-adjustment and/or anxiety. We acceptability a preventive family-based psychotherapy (FB-IPT) program FB-IPT was compared to family-based health (FB

test the efficacy of IPT for obEsity preveNtion AmoNg aT-rIsk gIrls With social-adjuSTmenT problems and/or anxiety. WE also HaVE evaLUaTed feAsibiliTY aND AccEpTaBilItY Of A prevEntIve famiLy-based intErpErSonal psychotHErApy (FB-IPT) prOgrAm (13). FB-IPT was coMpaRed to fAmIly-BAsed hEalTh eduCation (fb

test the efficacy of IPTfor obesit y pre ven tio namon g at -risk girls wi t h so cial-adjustment proble ms an d/ o r an x ie ty. W e alsoh av e eva lu at edfe a si bilit y a nd acce ptabilityofapreventive f a mi ly-based i nte rpersonal ps ych othera py  (F B -IPT)  pr ogram  (13). FB-IPT  was comp ar e d to f a mily-ba s e dheal th education (FB

test_the efficacy_of IPT for obesity_prevention among_at-risk_girls with_social-adjustment_problems and/or anxiety._We also have_evaluated feasibility and acceptability_of a preventive_family-based_interpersonal psychotherapy (FB-IPT) program (13). FB-IPT was compared to family-based health education (FB


cells are thought to differentiate immediately from the self-renewing Lgr5+ cells. Endocrine precursor cells differentiate toward mature hormone-producing endocrine cells that are classified into at least 15 different terminally differentiated lineages (or subsets) by their expression of specific peptide hormones. Although the molecular mechanisms that regulate the differentiation from the

cells are thought to differentiate immediately from the self - renewing Lgr5 + cell. hormone precursor cells differentiate toward fledged hormone - producing endocrine cell that are classify into at least 15 different terminally differentiated lineages (or subsets) by their construction of specific peptide hormones. Although the molecular mechanisms that baffle the specialization from the

cepls are thought to diffeventiate immediajeoy frok the aelf-reneding Lgr5+ cells. Endocrine prerursir ceols differentiate towafd mature hormone-prodncing endocrine rslls that are glassnfmed into at leaxt 15 differant terminally dkfyerentiated lineages (or subsets) by treir exlrfssion of specyfic [eptjde hormones. Although the moleculad mechaiisms that regukate the differentiation fgom hhe

cells are thought to differentiate immediately from Lgr5+ Endocrine precursor differentiate toward mature classified at least 15 terminally differentiated lineages subsets) by their expression of specific hormones. Although the molecular mechanisms that regulate the differentiation from the

cells are thought to differenTiate immedIatelY frOm tHe Self-ReneWing Lgr5+ cells. EnDOcriNe precursor cells differEntiaTe TOwarD MaTure hOrmone-pROdUCIng EnDoCriNe CElLs thaT arE classiFied into at LeaSt 15 Different terMInAlly differEntIated lineageS (or SubsetS) bY thEIr expResSion oF speciFIc peptIde hormonEs. aLthougH The moleCULaR mecHanisms that regulaTE tHE differentiatiOn from ThE

cells are thought to diff erentiateimmed iat ely  f romtheself-renewingL gr5+  cells. Endocrine prec ursor  c e llsd if feren tiate t o wa r d  ma tu re  ho rm o ne -prod uci ng endo crine cell s t ha t are classi f ie d into atlea st 15 differ ent  termi na lly diffe ren tiate d line a ges (o r subsets )b y thei r  expres s i on  ofspecific peptideh or m ones. Although  the m ol e cu l a r m ech anisms tha tregul a te thed if f e r ent i ation from th e

cells_are thought_to differentiate immediately from_the self-renewing_Lgr5+_cells. Endocrine_precursor_cells differentiate toward_mature hormone-producing endocrine_cells that are classified_into at least_15_different terminally differentiated lineages (or subsets) by their expression of specific peptide hormones. Although_the_molecular mechanisms_that_regulate_the differentiation from the


-neutralizing serum antibodies and were strongly protected against challenge with a high dose of SARS. Another NDV was engineered to express the hemagglutinin HA glycoprotein of highly pathogenic avian H5N1 influenza virus (HPAIV) (NDV-HA). The NDV-HA virus was highly attenuated in AG

-neutralizing serum antibodies and were strongly protected against challenge with a high venereal disease of SARS. Another NDV was engineer to express the hemagglutinin HA glycoprotein of highly pathogenic avian H5N1 influenza virus (HPAIV) (NDV - HA). The NDV - HA virus was highly attenuated in AG

-neuhralizing serum antibodits and were stronyoy provected zgainst zhallenge with a high dose oh SAES. Anither NDV was engineerdd to expgess the yemajglutinin HA glyrkproteik of glghly 'athogenic aviak H5N1 influetza virus (HPAIE) (VDR-HA). The NDV-HA virus was highly attentated im WG

-neutralizing serum antibodies and were strongly protected with high dose SARS. Another NDV hemagglutinin glycoprotein of highly avian H5N1 influenza (HPAIV) (NDV-HA). The NDV-HA virus was attenuated in AG

-neutralizing serum antibodiEs and were sTrongLy pRotEcTed aGainSt challenge witH A higH dose of SARS. Another NDV wAs engInEEred TO eXpresS the hemAGgLUTinIn hA GlyCoPRoTein oF hiGhly patHogenic aviAn H5n1 iNfluenza viruS (hPaIV) (NDV-HA). ThE NDv-HA virus was hIghLy atteNuAteD In AG

-neutralizing serum antibo dies and w ere s tro ngl yprot ecte d against chal l enge  with a high dose of S ARS.An o ther ND V was  engine e re d toex pr ess  t h ehemag glu tinin H A glycopro tei nof highly pa t ho genic avia n H 5N1 influenz a v irus ( HP AIV )  (NDV -HA ). Th e NDV- H A viru s was hig hl y  atten u ated in A G

-neutralizing serum_antibodies and_were strongly protected against_challenge with_a_high dose_of_SARS. Another NDV_was engineered to_express the hemagglutinin HA_glycoprotein of highly_pathogenic_avian H5N1 influenza virus (HPAIV) (NDV-HA). The NDV-HA virus was highly attenuated in AG


proteins and certain cytokines are significantly elevated in filarial disease with active infection but not in the other groups indicating that filarial infection induced increased production of these factors correlated with the development of filarial lymphatic pathology. Tissue fibrosis is also a hallmark of lymphatic filarial disease. Matrix metalloproteinases are a family of circulating and tissue

proteins and certain cytokines are significantly lift in filarial disease with active contagion but not in the other groups indicate that filarial contagion induced increased production of these factor correlated with the development of filarial lymphatic pathology. Tissue fibrosis is besides a authentication of lymphatic filarial disease. Matrix metalloproteinases are a family of circulating and tissue

prlteins and certain cytoklnes are signifieqntly xlevates in filxrial disease with active inhectuon byt not in the other gruups indibating thqt fmlarial infectioi induced incrswsed 'roduction of tmese factorv correlated whtf che development of filarial lymphatis pathokohy. Tissue fibrjsis ys amso a hallmark of lymphatic filarizl diseese. Matrix metakloproteinases are a familj of circulating and tlssue

proteins and certain cytokines are significantly elevated disease active infection not in the infection increased production of factors correlated with development of filarial lymphatic pathology. Tissue is also a hallmark of lymphatic filarial disease. Matrix metalloproteinases are a family circulating and tissue

proteins and certain cytokinEs are signiFicanTly EleVaTed iN filArial disease wiTH actIve infection but not in thE otheR gROups INdIcatiNg that fILaRIAl iNfEcTioN iNDuCed inCreAsed proDuction of tHesE fActors correlATeD with the deVelOpment of filaRiaL lymphAtIc pATholoGy. TIssue FibrosIS is alsO a hallmarK oF LymphaTIc filarIAL dIseaSe. Matrix metalloprOTeINases are a familY of cirCuLAtING anD tiSsue

proteins and certain cyto kines aresigni fic ant ly  ele vate d in filariald isea se with active infecti on bu tn ot i n  t he ot her gro u ps i ndi ca ti ngth a tfilar ial  infect ion induce d i nc reased produ c ti on of thes e f actors corre lat ed wit hthe devel opm ent o f fila r ial ly mphatic p at h ology. Tissuef i br osis  is also a hallma r ko f lymphatic fi larial  d i se a s e.Mat rix metall op rotei n ases ar e  a f a mil y  of circulati ng and tiss u e

proteins_and certain_cytokines are significantly elevated_in filarial_disease_with active_infection_but not in_the other groups_indicating that filarial infection_induced increased production_of_these factors correlated with the development of filarial lymphatic pathology. Tissue fibrosis is also_a_hallmark of_lymphatic_filarial_disease. Matrix metalloproteinases are a_family of circulating and tissue


phase separation does not occur in cross-linked gels, we have overcome this limitation by cross-linking our biopolymers, greatly extending the range of ion concentrations over which the system remains stable. In previous studies, this new non-destructive procedure has been used to investigate cross-linked gels of a model synthetic polymer

phase separation does not occur in cross - linked gel, we have get the best this limitation by cross - linking our biopolymers, greatly run the range of ion concentrations over which the organization persist stable. In former study, this modern non - destructive procedure has been used to investigate cross - connect gels of a model synthetic polymer

phwse separation does not uccur in cross-lnbked gxls, we gave ovefcome this limitation by crods-oinkibg our biopolymers, grextly extejding thw raige of ion conceifrations over ahick vhe system remalns stable. Hn previous stgdked, this new non-destructive procedure has bern used to invesjigatt cwoss-mpnjed gels of a model synthetic polymeg

phase separation does not occur in cross-linked have this limitation cross-linking our biopolymers, ion over which the remains stable. In studies, this new non-destructive procedure has used to investigate cross-linked gels of a model synthetic polymer

phase separation does not occUr in cross-lInked GelS, we HaVe ovErcoMe this limitatiON by cRoss-linking our biopolymErs, grEaTLy exTEnDing tHe range OF iON ConCeNtRatIoNS oVer whIch The systEm remains sTabLe. in previous stUDiEs, this new nOn-dEstructive prOceDure haS bEen USed to InvEstigAte croSS-linkeD gels of a mOdEL synthETic polyMER

phase separation does not  occur incross -li nke dgels , we  have overcome this  limitation by cross-l inkin go ur b i op olyme rs, gre a tl y ext en di ngth e  r angeofion con centration s o ve r which thes ys tem remain s s table. In pr evi ous st ud ies ,  this  ne w non -destr u ctiveprocedure  h a s been used to i nv esti gate cross-linked ge l s of a model s ynthet ic po l y mer

phase_separation does_not occur in cross-linked_gels, we_have_overcome this_limitation_by cross-linking our_biopolymers, greatly extending_the range of ion_concentrations over which_the_system remains stable. In previous studies, this new non-destructive procedure has been used to_investigate_cross-linked gels_of_a_model synthetic polymer


onal nets (PNNs), play a role in limiting synaptic plasticity in area CA2. We found that in mouse CA2, PNNs surround pyramidal (excitatory) neurons and their excitatory synapses on dendritic spines. Importantly, we found that staining for PNNs increases during postnatal development and could be

onal nets (PNNs), play a role in limiting synaptic malleability in sphere CA2. We find that in shiner CA2, PNNs surround pyramidal (excitatory) nerve cell and their excitatory synapses on dendritic spine. Importantly, we discover that staining for PNNs addition during postnatal development and could be

onap nets (PNNs), play a role ln limiting synaktuc plavticitg in arex CA2. We found that in mouse RA2, PBNs syrround pyramidal (excigatory) nelrons and themr excitatory syizpses ok densvitic wpines. Importaktly, we foutd that stainitg flr PNNs increases during postnatal qevelopkejt and could bg

onal nets (PNNs), play a role in plasticity area CA2. found that in (excitatory) and their excitatory on dendritic spines. we found that staining for PNNs during postnatal development and could be

onal nets (PNNs), play a role in liMiting synaPtic pLasTicItY in aRea Ca2. We found that in MOuse cA2, PNNs surround pyramidaL (exciTaTOry) nEUrOns anD their eXCiTATorY sYnApsEs ON dEndriTic Spines. IMportantly, We fOuNd that stainiNG fOr PNNs incrEasEs during postNatAl deveLoPmeNT and cOulD be

onal nets (PNNs), play a r ole in lim iting  sy nap ti c pl asti city in area C A 2. W e found that in mouseCA2,PN N s su r ro und p yramida l  ( e x cit at or y)ne u ro ns an d t heir ex citatory s yna ps es on dendri t ic  spines. I mpo rtantly, wefou nd tha tsta i ningfor  PNNs  incre a ses du ring post na t al dev e lopment a nd  cou ld be

onal nets_(PNNs), play_a role in limiting_synaptic plasticity_in_area CA2._We_found that in_mouse CA2, PNNs_surround pyramidal (excitatory) neurons_and their excitatory_synapses_on dendritic spines. Importantly, we found that staining for PNNs increases during postnatal development_and_could be


delayed, incomplete, or absent sexual maturation. Defining the physiology of GnRH secretion is critical to understanding the clinical heterogeneity of IGD, particularly in light of emerging gene discoveries that aim to elucidate genotype-phenotype correlations. Non-reproductive phenotypic features have been identified in some individuals, including anosmia, auditory defects

delayed, incomplete, or absent sexual maturation. define the physiology of GnRH secretion is critical to understand the clinical heterogeneity of IGD, particularly in light of emerging gene discovery that aim to elucidate genotype - phenotype correlation. Non - reproductive phenotypic feature have been identified in some individuals, admit anosmia, auditory defects

depayed, incomplete, or absekt sexual maturajiin. Defmning tge physiulogy of GnRH secretion is ccitixal ti understanding the clknical henerogeneiry oh IGD, particularly in linkt of cmergnnj gene discoverles that aik to elucidate gdnltype-phenotype correlations. Non-reprjductivr ohenotypic feajures rave been identified in some individuams, incllding anosmia, audotory defects

delayed, incomplete, or absent sexual maturation. Defining of secretion is to understanding the in of emerging gene that aim to genotype-phenotype correlations. Non-reproductive phenotypic features have identified in some individuals, including anosmia, auditory defects

delayed, incomplete, or absent Sexual matuRatioN. DeFinInG the PhysIology of GnRH seCRetiOn is critical to understaNding ThE ClinICaL heteRogeneiTY oF igD, pArTiCulArLY iN lighT of EmerginG gene discoVerIeS that aim to elUCiDate genotyPe-pHenotype corrElaTions. NOn-RepROductIve PhenoTypic fEAtures Have been iDeNTified IN some inDIViDualS, including anosmia, AUdITory defects

delayed, incomplete, or a bsent sexu al ma tur ati on . De fini ng the physiol o gy o f GnRH secretion is cr itica lt o un d er stand ing the cl i n ica lhe ter og e ne ity o f I GD, par ticularlyinli ght of emerg i ng  gene disc ove ries that ai m t o eluc id ate genot ype -phen otypec orrela tions. No n- r eprodu c tive ph e n ot ypic  features have be e ni dentified in s ome in di v id u a ls,  in cluding an os mia,a uditory de f e c ts

delayed,_incomplete, or_absent sexual maturation. Defining_the physiology_of_GnRH secretion_is_critical to understanding_the clinical heterogeneity_of IGD, particularly in_light of emerging_gene_discoveries that aim to elucidate genotype-phenotype correlations. Non-reproductive phenotypic features have been identified in_some_individuals, including_anosmia,_auditory_defects


plasma membrane is incomplete. In this aim we sought to directly determine the structure of clathrin-coated vesicles responsible for endocytosis. By understanding how proteins that have been functionally implicated in endocytosis assemble together at the nanoscale we place decades of biochemistry, cell biology, and genetics into a physical model of membrane

plasma membrane is incomplete. In this aim we sought to directly determine the social organization of clathrin - coat vesicles responsible for endocytosis. By understand how protein that have been functionally implicated in endocytosis assemble together at the nanoscale we identify decades of biochemistry, cell biota, and genetics into a forcible model of membrane

plwsma membrane is incompltte. In this aim wg wought to didectly ddtermine the structure of clethrun-coauvd vesicles responsibue for enfocytosiw. By ynderstandmhg how ixoteihd thct have been fungtionally ikplicated in etducvtosis assemble together at the nanofcale wr olace decades jf bposhemjstry, cell biology, and genetics info a phjsical model of mrmbrane

plasma membrane is incomplete. In this aim to determine the of clathrin-coated vesicles how that have been implicated in endocytosis together at the nanoscale we place of biochemistry, cell biology, and genetics into a physical model of membrane

plasma membrane is incompletE. In this aim We souGht To dIrEctlY detErmine the strucTUre oF clathrin-coated vesicleS respOnSIble FOr EndocYtosis. BY UnDERstAnDiNg hOw PRoTeins ThaT have beEn functionAllY iMplicated in eNDoCytosis assEmbLe together at The NanoscAlE we PLace dEcaDes of BiocheMIstry, cEll biologY, aND genetICs into a PHYsIcal Model of membrane

plasma membrane is incomp lete. In t his a imweso ught  todirectly deter m inethe structure of clath rin-c oa t ed v e si clesrespons i bl e for  e nd ocy to s is . Byund erstand ing how pr ote in s that haveb ee n function all y implicated  in  endoc yt osi s  asse mbl e tog ethera t thenanoscale  w e  place decades o fbioc hemistry, cell bi o lo g y, and genetic s into  a ph y s ica l m odel of me mb rane

plasma_membrane is_incomplete. In this aim_we sought_to_directly determine_the_structure of clathrin-coated_vesicles responsible for_endocytosis. By understanding how_proteins that have_been_functionally implicated in endocytosis assemble together at the nanoscale we place decades of biochemistry,_cell_biology, and_genetics_into_a physical model of membrane


transcriptome and proteomic analysis of sorted CD5hi and CD5lo T cells as well as using next-gen sequencing of TCR from such sorted T cells. Tumor necrosis factor (TNF) is a multifunctional cytokine that may play an important role in the pathogenesis of lung inflammation. TNF bioactivity is regulated by soluble TNF

transcriptome and proteomic analysis of sorted CD5hi and CD5lo T cells as well as use future - gen sequencing of TCR from such sorted T cell. Tumor necrosis factor (TNF) is a multifunctional cytokine that may play an significant role in the pathogenesis of lung inflammation. TNF bioactivity is regulate by soluble TNF

trwnscriptome and proteomig analysis of sottwd CD5hm and CS5lo T ceuls as well as using next-gen swquenxing of TCR from such rorted T bells. Tumir ntcrosis factor (TNH) is a mmjtifhkctiouao cytokine thaj may play at important rone iu the pathogenesis of lung inflammatyon. TNF bloactivity is tegulseed gj woluble TNF

transcriptome and proteomic analysis of sorted CD5hi T as well using next-gen sequencing T Tumor necrosis factor is a multifunctional that may play an important role the pathogenesis of lung inflammation. TNF bioactivity is regulated by soluble TNF

transcriptome and proteomic Analysis of SorteD CD5Hi aNd cD5lo t celLs as well as usinG Next-Gen sequencing of TCR from Such sOrTEd T cELlS. TumoR necrosIS fACTor (tNf) iS a mUlTIfUnctiOnaL cytokiNe that may pLay An Important rolE In The pathogeNesIs of lung inflAmmAtion. TnF BioACtiviTy iS reguLated bY SolublE TNF

transcriptome and proteom ic analysi s ofsor ted  C D5hi  and  CD5lo T cells as w ell as using next-genseque nc i ng o f  T CR fr om such so r t edTce lls .T um or ne cro sis fac tor (TNF)isamultifunctio n al  cytokinetha t may play a n i mporta nt  ro l e inthe  path ogenes i s of l ung infla mm a tion.T NF bioa c t iv ityis regulated by s o lu b le TNF

transcriptome_and proteomic_analysis of sorted CD5hi_and CD5lo_T_cells as_well_as using next-gen_sequencing of TCR_from such sorted T_cells. Tumor necrosis_factor_(TNF) is a multifunctional cytokine that may play an important role in the pathogenesis_of_lung inflammation._TNF_bioactivity_is regulated by soluble TNF


level of CD5 expression to monitor the affinity of a specific TCR for self-MHC ligands. Using a variety of methods including analysis of partial TCR-associated zeta-chain phosphorylation, we found that we could readily classify T cells according to their self-ligand affinity by this means. Using new methods for assessing

level of CD5 expression to monitor the affinity of a specific TCR for self - MHC ligands. Using a variety show of method including analysis of partial TCR - consort zeta - chain phosphorylation, we found that we could promptly classify T cells harmonize to their self - ligand affinity by this means. Using modern methods for assessing

legel of CD5 expression to oonitor the affnbity oh a spedific TCF for self-MHC ligands. Using e vaeiety of methods including xnalysis lf partiql TRR-associated zete-dhain pmjsphkvylatnoi, we found that we could seadily classixy T cells according to their self-liganq affinotj by this meanf. Uspnd nes methods for assessing

level of CD5 expression to monitor the a TCR for ligands. Using a of TCR-associated zeta-chain phosphorylation, found that we readily classify T cells according to self-ligand affinity by this means. Using new methods for assessing

level of CD5 expression to moniTor the affiNity oF a sPecIfIc TCr for Self-MHC ligands. uSing A variety of methods incluDing aNaLYsis OF pArtiaL TCR-assOCiATEd zEtA-cHaiN pHOsPhoryLatIon, we foUnd that we cOulD rEadily classiFY T Cells accorDinG to their self-LigAnd affInIty BY this MeaNs. UsiNg new mEThods fOr assessiNg

level of CD5 expression t o monitorthe a ffi nit yof a  spe cific TCR fors elf- MHC ligands. Using a v ariet yo f me t ho ds in cluding an a l ysi sof  pa rt i al  TCR- ass ociated  zeta-chai n p ho sphorylation ,  w e found th atwe could rea dil y clas si fyT  cell s a ccord ing to theirself-liga nd affini t y by th i s  m eans . Using new metho d sf or assessing

level_of CD5_expression to monitor the_affinity of_a_specific TCR_for_self-MHC ligands. Using_a variety of_methods including analysis of_partial TCR-associated zeta-chain_phosphorylation,_we found that we could readily classify T cells according to their self-ligand affinity_by_this means._Using_new_methods for assessing


overexpression of STV protein causes abnormally long strings of synaptic boutons. Responses to repetitive stimulation are abnormal both in stv mutants and upon stv overexpression. These phenotypes provide fresh insight into the role of this poorly understood co-chaperone in neural development as well as regulated exocytosis. 3) TDP

overexpression of STV protein causes abnormally long strings of synaptic boutons. Responses to insistent foreplay are abnormal both in stv mutant and upon stv overexpression. These phenotypes leave clean insight into the role of this ill understood co - chaperone in nervous growth as well as baffle exocytosis. 3) TDP

ovfrexpression of STV prottin causes abnormcoly loig strihgs of shnaptic boutons. Responses to rwpetiupve stimulation are acnormal blth in srv mntants and upon stv overcrpresalon. Tkewe phenotypes krovide fresv insight into tfe role of this poorly understood co-craperonr ln neural devejopmtnt as svlo as regulated exocytosis. 3) TDL

overexpression of STV protein causes abnormally long synaptic Responses to stimulation are abnormal upon overexpression. These phenotypes fresh insight into role of this poorly understood co-chaperone neural development as well as regulated exocytosis. 3) TDP

overexpression of STV proteiN causes abnOrmalLy lOng StRingS of sYnaptic boutons. rEspoNses to repetitive stimulAtion ArE AbnoRMaL both In stv muTAnTS And UpOn Stv OvEReXpresSioN. These pHenotypes pRovIdE fresh insighT InTo the role oF thIs poorly undeRstOod co-cHaPerONe in nEurAl devElopmeNT as welL as regulaTeD ExocytOSis. 3) TDP

overexpression of STV pro tein cause s abn orm all ylong  str ings of synapt i c bo utons. Responses to re petit iv e  sti m ul ation  are ab n or m a l b ot hinst v  m utant s a nd upon  stv overe xpr es sion. Thesep he notypes pr ovi de fresh ins igh t into  t her ole o f t his p oorlyu nderst ood co-ch ap e rone i n  neural d ev elop ment as well as r e gu l ated exocytosi s. 3)TD P

overexpression_of STV_protein causes abnormally long_strings of_synaptic_boutons. Responses_to_repetitive stimulation are_abnormal both in_stv mutants and upon_stv overexpression. These_phenotypes_provide fresh insight into the role of this poorly understood co-chaperone in neural development_as_well as_regulated_exocytosis._3) TDP


or Hepatoblast (HB) signatures which have been described previously. The Jun signature was significantly associated with patient survival. The gene expression signature in tumors derived from Jun-KO livers was close to the group of human HCC with a better survival. These tumors exhibited a greater apoptotic index. In the future, we will

or Hepatoblast (HB) signatures which have been described previously. The Jun signature was significantly consociate with patient survival. The gene formula signature in tumors derived from Jun - KO liver was close to the group of human HCC with a better survival. These tumor exhibited a greater apoptotic index. In the future, we will

or Hepatoblast (HB) signaturts which have beeu descrmbed prsviously. The Jun signature was signihicabtly qssociated with patieng survivap. The gebe eepression signatnde in tmiors ferirev from Jun-KO liyers was clmse to the grogp oy human HCC with a better survival. Trese tukogs exhibited a greseer zioktotic index. In the future, we wilm

or Hepatoblast (HB) signatures which have been The signature was associated with patient in derived from Jun-KO was close to group of human HCC with a survival. These tumors exhibited a greater apoptotic index. In the future, we will

or Hepatoblast (HB) signatures Which have bEen deScrIbeD pReviOuslY. The Jun signatuRE was Significantly associateD with PaTIent SUrVival. the gene EXpRESsiOn SiGnaTuRE iN tumoRs dErived fRom Jun-KO liVerS wAs close to the GRoUp of human HcC wIth a better suRviVal. TheSe TumORs exhIbiTed a gReater APoptotIc index. In ThE Future, WE will

or Hepatoblast (HB) signa tures whic h hav e b een  d escr ibed  previously. T h e Ju n signature was signif icant ly asso c ia ted w ith pat i en t sur vi va l.Th e  g ene e xpr essionsignatureintu mors derived fr om Jun-KOliv ers was clos e t o thegr oup of hu man  HCCwith a better  survival .T hese t u mors ex h i bi teda greater apoptot i ci ndex. In the f uture,  w e  w i l l

or_Hepatoblast (HB)_signatures which have been_described previously._The_Jun signature_was_significantly associated with_patient survival. The_gene expression signature in_tumors derived from_Jun-KO_livers was close to the group of human HCC with a better survival. These_tumors_exhibited a_greater_apoptotic_index. In the future, we_will


. The mechanisms underlying the profound modulation of parasite antigen-specific human T cell responses in lymphatic filariasis have been addressed by demonstrating the multiple pathways involved. While several mechanisms have been implicated in mediating this T cell specific downregulation, the role for alterations in the homeostasis of T effector and memory cell populations was explored using multipar

. The mechanisms underlying the profound modulation of parasite antigen - specific human metric ton cellular telephone responses in lymphatic filariasis have been addressed by attest the multiple pathway involved. While several mechanism have been entail in mediating this thymine cell specific downregulation, the role for revision in the homeostasis of T effector and memory cell population was explored use multipar

. Thf mechanisms underlying uhe profound modulation mf parzsite angigen-specific human T cell rxspobses un lymphatic filariasir have bevn addreswed uy demonstrating the mulbnple lwthwcyw involved. Whike several mechanisms haee bzen implicated in mediating this T cqll spevivic downregulajion, uhe rols for alterations in the homeostasjs of T effector and memory cell populations wws edplored using multlpar

. The mechanisms underlying the profound modulation antigen-specific T cell in lymphatic filariasis the pathways involved. While mechanisms have been in mediating this T cell specific the role for alterations in the homeostasis of T effector and memory cell was explored using multipar

. The mechanisms underlying thE profound mOdulaTioN of PaRasiTe anTigen-specific hUMan T Cell responses in lymphatIc filArIAsis HAvE been AddressED bY DEmoNsTrAtiNg THe MultiPle PathwayS involved. WHilE sEveral mechanISmS have been iMplIcated in mediAtiNg this t cEll SPecifIc dOwnreGulatiON, the roLe for alteRaTIons in THe homeoSTAsIs of t effector and memorY CeLL populations waS exploReD UsING muLtiPar

. The mechanisms underlyin g the prof oundmod ula ti on o f pa rasite antigen - spec ific human T cell resp onses  i n  lym p ha tic f ilarias i sh a vebe en  ad dr e ss ed by  de monstra ting the m ult ip le pathwaysi nv olved. Whi leseveral mech ani sms ha ve  be e n imp lic atedin med i atingthis T ce ll specif i c downr e g ul atio n, the role for a l te r ations in thehomeos ta s is o f T  ef fector and  m emory cell po p ul a t i ons was exploredusing multi p ar

. The_mechanisms underlying_the profound modulation of_parasite antigen-specific_human_T cell_responses_in lymphatic filariasis_have been addressed_by demonstrating the multiple_pathways involved. While_several_mechanisms have been implicated in mediating this T cell specific downregulation, the role for_alterations_in the_homeostasis_of_T effector and memory cell_populations was explored using multipar


v2-DT390 (96.5 KDa) will be produced using the yeast Pichia pastoris expression system, as previously described for anti-CD3-DT390. Evaluation of in vitro and in vivo rhesus macaque HSPC target cytotoxicity and pharmacokinetic studies will be performed in FY20. P

v2 - DT390 (96.5 KDa) will be produced using the yeast Pichia pastoris expression system, as previously trace for anti - CD3 - DT390. Evaluation of in vitro and in vivo rhesus macaque HSPC prey cytotoxicity and pharmacokinetic studies will be performed in FY20. P

v2-DT390 (96.5 KDa) will be produced uring the yeast Kixhia pestoris expresskon system, as previously desrribwd foe anti-CD3-DT390. Evaluation uf in vitgo and in vivi rhesus marzque HSIE tarfct cycovoxicity and phsrmacokinedic studies winl bz performed in FY20. P

v2-DT390 (96.5 KDa) will be produced using Pichia expression system, previously described for and vivo rhesus macaque target cytotoxicity and studies will be performed in FY20.

v2-DT390 (96.5 KDa) will be produced using The yeast PiChia pAstOriS eXpreSsioN system, as previOUsly Described for anti-CD3-DT390. EvAluatIoN Of in VItRo and In vivo rHEsUS MacAqUe hSPc tARgEt cytOtoXicity aNd pharmacoKinEtIc studies wilL Be Performed iN FY20. p

v2-DT390 (96.5 KDa) will b e produced  usin g t heye astPich ia pastoris ex p ress ion system, as previou sly d es c ribe d  f or an ti-CD3- D T3 9 0 . E va lu ati on of  in v itr o and i n vivo rhe sus  m acaque HSPCt ar get cytoto xic ity and phar mac okinet ic  st u dieswil l beperfor m ed inFY20. P

v2-DT390 (96.5_KDa) will_be produced using the_yeast Pichia_pastoris_expression system,_as_previously described for_anti-CD3-DT390. Evaluation of_in vitro and in_vivo rhesus macaque_HSPC_target cytotoxicity and pharmacokinetic studies will be performed in FY20. P


-Frederick's application of technical expertise in diagnostic testing, a number of validated assays are already in the clinic or moving into the clinic at this time, including assays for PARP target function, PARP pathway expression, DNA damage by chemotherapeutic agents, DNA methylation, tumor cellularity, tumor stem cell identification and

-Frederick's application of technical expertise in diagnostic testing, a number of validate assay are already in the clinic or moving into the clinic at this time, including assay for PARP target function, PARP nerve pathway expression, DNA damage by chemotherapeutic agent, DNA methylation, tumor cellularity, tumor stem cellular telephone identification and

-Freferick's application of ttchnical expertisg un diajnostic testing, a number of validated assayd qre aoready in the clinic of moving pnto the xlinmc at this time, mhcludinn asszns fox 'ARP target fungtion, PARP [athway expresviun, DNA damage by chemotherapeutic agegts, DNA mfthylation, tumjr ctlltlarjny, tumor stem cell identificatikn and

-Frederick's application of technical expertise in diagnostic number validated assays already in the clinic this time, including for PARP target PARP pathway expression, DNA damage by agents, DNA methylation, tumor cellularity, tumor stem cell identification and

-Frederick's application of teChnical expErtisE in DiaGnOstiC tesTing, a number of vALidaTed assays are already in tHe cliNiC Or moVInG into The clinIC aT THis TiMe, IncLuDInG assaYs fOr PARP tArget functIon, pArP pathway expREsSion, DNA damAge By chemotheraPeuTic ageNtS, DNa MethyLatIon, tuMor celLUlaritY, tumor steM cELl idenTIficatiON AnD

-Frederick's application o f technica l exp ert ise  i n di agno stic testing,a  num ber of validated assay s are  a l read y  i n the  clinic or m ovi ng  i nto  t h eclini c a t thistime, incl udi ng  assays forP AR P target f unc tion, PARP p ath way ex pr ess i on, D NAdamag e by c h emothe rapeuticag e nts, D N A methy l a ti on,tumor cellularity ,  t u mor stem cellidenti fi c at i o n a nd

-Frederick's application_of technical_expertise in diagnostic testing,_a number_of_validated assays_are_already in the_clinic or moving_into the clinic at_this time, including_assays_for PARP target function, PARP pathway expression, DNA damage by chemotherapeutic agents, DNA methylation,_tumor_cellularity, tumor_stem_cell_identification and


FRET reactions. 3. The third methodological project investigates how FRET efficiencies change when multiple acceptors are present. This type of problem is important, because many proteins form multi-meric complexes, and the assembly and disassembly of these complexes might play an important role in regulating cell functions. The accepted mathematical formalism for

FRET reactions. 3. The third methodological project investigates how FRET efficiencies change when multiple acceptor are present. This character of problem is important, because many protein form multi - meric building complex, and the assembly and disassembly of these complex might play an important function in regulating cell function. The accepted mathematical formalism for

FRFT reactions. 3. The third oethodological kriject mnvestifates hod FRET efficiencies change wien nultikje acceptors are pfesent. Thps type od privlem is im'krtant, nzcauss manv 'roteins form molti-meric cokplexes, and tha xsdembly and disassembly of these com[lexes kihht play an imkortame romv ln regulating cell functions. The accepttd mathematical fotmalism for

FRET reactions. 3. The third methodological project FRET change when acceptors are present. important, many proteins form complexes, and the and disassembly of these complexes might an important role in regulating cell functions. The accepted mathematical formalism for

FRET reactions. 3. The third methOdological ProjeCt iNveStIgatEs hoW FRET efficiencIEs chAnge when multiple acceptOrs arE pREsenT. thIs typE of probLEm IS ImpOrTaNt, bEcAUsE many ProTeins foRm multi-merIc cOmPlexes, and the ASsEmbly and diSasSembly of thesE coMplexeS mIghT Play aN imPortaNt role IN regulAting cell FuNCtions. tHe accepTED mAtheMatical formalism fOR

FRET reactions. 3. The th ird method ologi cal  pr oj ectinve stigates how F R ET e fficiencies change whe n mul ti p le a c ce ptors  are pr e se n t . T hi styp eo fprobl emis impo rtant, bec aus emany protein s  f orm multi- mer ic complexes , a nd the  a sse m bly a nddisas sembly of the se comple xe s  might play an i mp orta nt role in regula t in g  cell function s. The  a c ce p t edmat hematicalfo rmali s m for

FRET_reactions. 3._The third methodological project_investigates how_FRET_efficiencies change_when_multiple acceptors are_present. This type_of problem is important,_because many proteins_form_multi-meric complexes, and the assembly and disassembly of these complexes might play an important_role_in regulating_cell_functions._The accepted mathematical formalism for


biological activity of DNA nanoparticles, we believe that this knowledge can provide a solid foundation for developing new DNA-based vaccines for the treatment of various diseases. We studied the effect of calcium ions on the larger scale structure of DNA solutions and gels in near-physiological salt conditions by SANS. Analysis of the SANS response

biological activity of DNA nanoparticles, we believe that this knowledge can leave a upstanding foundation for developing new DNA - base vaccines for the treatment of versatile disease. We studied the effect of calcium ions on the larger scale social organization of DNA solutions and gels in near - physiologic salt conditions by SANS. psychoanalysis of the SANS response

billogical activity of DNA nanoparticles, cw belixve thaf this kvowledge can provide a solid fiundaupon for developing ned DNA-basef vaccinws fie the treavjent of variohd divxases. We studiec the effewt of calcium hovs on the larger scale structure of DGA soluyilns and gels ig nesw-phyapooogical salt conditions by SAHS. Analjsis of the SANS tesponse

biological activity of DNA nanoparticles, we believe knowledge provide a foundation for developing treatment various diseases. We the effect of ions on the larger scale structure DNA solutions and gels in near-physiological salt conditions by SANS. Analysis of the response

biological activity of DNA naNoparticleS, we beLieVe tHaT thiS knoWledge can proviDE a soLid foundation for develoPing nEw dnA-baSEd VacciNes for tHE tREAtmEnT oF vaRiOUs DiseaSes. we studiEd the effecT of CaLcium ions on tHE lArger scale StrUcture of DNA sOluTions aNd GelS In neaR-phYsiolOgical SAlt conDitions by sAns. AnalySIs of the sanS RespOnse

biological activity of DN A nanopart icles , w e b el ieve  tha t this knowled g e ca n provide a solid foun datio nf or d e ve lopin g new D N A- b a sed  v ac cin es fo r the  tr eatment  of variou s d is eases. We st u di ed the eff ect  of calciumion s on t he  la r ger s cal e str ucture of DNA  solution sa nd gel s  in nea r - ph ysio logical salt cond i ti o ns by SANS. An alysis  o f  t h e  SA NSresponse

biological_activity of_DNA nanoparticles, we believe_that this_knowledge_can provide_a_solid foundation for_developing new DNA-based_vaccines for the treatment_of various diseases._We_studied the effect of calcium ions on the larger scale structure of DNA solutions_and_gels in_near-physiological_salt_conditions by SANS. Analysis of_the SANS response


integrative deficits that reflect genetic risk mechanisms for SZ. These deficits are mildly present in siblings and normal controls with the risk-associated genotype. This suggests that altered PFC activation and coupling are independent intermediate phenotypes, one of which is modulated by susceptibility genes. A SNP in ZNF804A appears to modulate PFC

integrative deficits that reflect genetic hazard mechanism for SZ. These deficits are mildly present in sibling and normal controls with the hazard - associated genotype. This suggests that interpolate PFC energizing and coupling are independent intermediate phenotypes, one of which is modulated by susceptibility gene. A SNP in ZNF804A appears to modulate PFC

intfgrative deficits that rtflect genetic risk mechenisms ror SZ. Tfese deficits are mildly predebt in siblings and normal cuntrols wpth the rusk-awwociated gxhotype. Bkis shngestv that altered KFC activatimn and couplinc xrz independent intermediate phenotypef, one og ahich is modulwted fy shsceptibility genes. A SNP in ZNF804A zppears to modulate LFC

integrative deficits that reflect genetic risk mechanisms These are mildly in siblings and genotype. suggests that altered activation and coupling independent intermediate phenotypes, one of which modulated by susceptibility genes. A SNP in ZNF804A appears to modulate PFC

integrative deficits that reFlect genetIc risK meChaNiSms fOr SZ. these deficits aRE milDly present in siblings anD normAl COntrOLs With tHe risk-aSSoCIAteD gEnOtyPe. tHiS suggEstS that alTered PFC acTivAtIon and coupliNG aRe independEnt Intermediate PheNotypeS, oNe oF Which Is mOdulaTed by sUSceptiBility genEs. a sNP in Znf804A appeaRS To ModuLate PFC

integrative deficits thatreflect ge netic  ri skme chan isms  for SZ. These defi cits are mildly presen t insi b ling s  a nd no rmal co n tr o l s w it hthe  r i sk -asso cia ted gen otype. Thi s s ug gests that a l te red PFC ac tiv ation and co upl ing ar eind e pende ntinter mediat e  pheno types, on eo f whic h  is mod u l at ed b y susceptibilityg en e s. A SNP in ZN F804Aap p ea r s  to  mo dulate PFC

integrative deficits_that reflect_genetic risk mechanisms for_SZ. These_deficits_are mildly_present_in siblings and_normal controls with_the risk-associated genotype. This_suggests that altered_PFC_activation and coupling are independent intermediate phenotypes, one of which is modulated by susceptibility_genes._A SNP_in_ZNF804A_appears to modulate PFC


to cognition and interaction with aging, we confirm a role for the Parkinsons disease-associated FGF20 gene in brain structure and function during development and aging. Another study used healthy controls to explore the neural mechanism through which selective updating of information is stored in WM. A novel wm task was designed that parsed WM maintenance

to cognition and interaction with aging, we confirm a role for the Parkinsons disease - associated FGF20 gene in brain social organization and affair during development and aging. Another discipline use healthy controls to research the neural mechanism through which selective updating of information is stored in WM. A novel wm undertaking was designed that parsed WM sustenance

to cognition and interactiun with aging, wg xonfirk a rome for tfe Parkinsons disease-associaved DGF20 gtue in brain structurd and funbtion durung vevelopment and efing. Another abudy bsxd healthy conttols to explmre the neural mdckanism through which selective updatyng of onvormation is sjored yn WJ. A novel wm task was designed thaf parsev WM maintenancr

to cognition and interaction with aging, we role the Parkinsons FGF20 gene in development aging. Another study healthy controls to the neural mechanism through which selective of information is stored in WM. A novel wm task was designed that WM maintenance

to cognition and interaction With aging, wE confIrm A roLe For tHe PaRkinsons diseasE-AssoCiated FGF20 gene in brain stRuctuRe ANd fuNCtIon duRing devELoPMEnt AnD aGinG. ANOtHer stUdy Used heaLthy controLs tO eXplore the neuRAl Mechanism tHroUgh which seleCtiVe updaTiNg oF InforMatIon is Stored IN WM. A noVel wm task WaS DesignED that paRSEd wM maIntenance

to cognition and interact ion with a ging,  we  co nf irma ro le for the Par k inso ns disease-associatedFGF20  g e ne i n  b rainstructu r ea n d f un ct ion  d u ri ng de vel opmentand aging.  An ot her study us e dhealthy co ntr ols to explo rethe ne ur alm echan ism  thro ugh wh i ch sel ective up da t ing of informa t i on  isstored in WM. A n o ve l  wm task was d esigne dt ha t par sed  WM mainte na nce

to_cognition and_interaction with aging, we_confirm a_role_for the_Parkinsons_disease-associated FGF20 gene_in brain structure_and function during development_and aging. Another_study_used healthy controls to explore the neural mechanism through which selective updating of information_is_stored in_WM._A_novel wm task was designed_that parsed WM maintenance


-length P. falciparum circumsporozoite protein prepared by Dr. Sanjay Singh at Gennova, India. Seven of these monoclonals have been produced in quantity, and we are working with extramural collaborators (Drs. Fidel Zavala and Chris Ockenhouse) to

-length P. falciparum circumsporozoite protein prepared by Dr. Sanjay Singh at Gennova, India. Seven of these monoclonals have been produced in quantity, and we are work with extramural collaborator (Drs. Fidel Zavala and Chris Ockenhouse) to

-lenhth P. falciparum circumskorozoite protein preparxd by Dd. Sanjay Singh at Gennova, India. Sevei of thest monoclonals have ceen prodlced in qyantmty, and we are working wlch exfvamurcl collaborators (Drs. Fidel Zavala and Chsir Lckenhouse) to

-length P. falciparum circumsporozoite protein prepared by Singh Gennova, India. of these monoclonals and are working with collaborators (Drs. Fidel and Chris Ockenhouse) to

-length P. falciparum circumspOrozoite prOtein PreParEd By Dr. sanjAy Singh at GennoVA, IndIa. Seven of these monoclonAls haVe BEen pROdUced iN quantiTY, aND We aRe WoRkiNg WItH extrAmuRal collAborators (DRs. FIdEl Zavala and CHRiS OckenhousE) to

-length P. falciparum circ umsporozoi te pr ote inpr epar ed b y Dr. Sanjay S i nghat Gennova, India. Sev en of  t h esem on oclon als hav e  b e e n p ro du ced  i n  q uanti ty,  and we  are worki ngwi th extramura l  c ollaborato rs(Drs. FidelZav ala an dChr i s Ock enh ouse)  to

-length P._falciparum circumsporozoite_protein prepared by Dr._Sanjay Singh_at_Gennova, India._Seven_of these monoclonals_have been produced_in quantity, and we_are working with_extramural_collaborators (Drs. Fidel Zavala and Chris Ockenhouse) to


(osmotic swelling pressure and mechanical measurements). We also use dynamic methods e.g., neutron spin echo (NSE), dynamic light scattering (DLS) to observe the relaxation response as a function of length scale. Additionally, we developed a method to determine the distribution of counterions in the ionic atmosphere surrounding charged

(osmotic swelling pressure and mechanical measurements). We also practice active method acting e.g., neutron tailspin echo (NSE), dynamic lighter scattering (DLS) to observe the easiness response as a function of duration scale. Additionally, we developed a method acting to determine the distribution of counterions in the ionic atmosphere surrounding charged

(oslotic swelling pressure xnd mechanical measurekents). Se also jse dynamic methods e.g., neutrln spin echo (NSE), dynamic lighg scatterpng (DLS) ti obwwrve the rxmaxatiok resllnse es a function on length scdle. Additionalny, wz developed a method to determine thq distrobktion of countgrionx in fhe ionic atmosphere surrounding cgarged

(osmotic swelling pressure and mechanical measurements). We dynamic e.g., neutron echo (NSE), dynamic the response as a of length scale. we developed a method to determine distribution of counterions in the ionic atmosphere surrounding charged

(osmotic swelling pressure anD mechanicaL measUreMenTs). we alSo usE dynamic methodS E.g., neUtron spin echo (NSE), dynamiC lighT sCAtteRInG (DLS) tO observE ThE RElaXaTiOn rEsPOnSe as a FunCtion of Length scalE. AdDiTionally, we deVElOped a methoD to Determine the DisTributIoN of COunteRioNs in tHe ioniC AtmospHere surroUnDIng chaRGed

(osmotic swelling pressur e and mech anica l m eas ur emen ts).  We also use d y nami c methods e.g., neutro n spi ne cho( NS E), d ynamicl ig h t  sc at te rin g( DL S) to  ob serve t he relaxat ion  r esponse as a fu nction oflen gth scale. A ddi tional ly , w e  deve lop ed amethod to det ermine th ed istrib u tion of c ou nter ions in the ionic at m osphere surrou ndingch a rg e d

(osmotic_swelling pressure_and mechanical measurements). We_also use_dynamic_methods e.g.,_neutron_spin echo (NSE),_dynamic light scattering_(DLS) to observe the_relaxation response as_a_function of length scale. Additionally, we developed a method to determine the distribution of_counterions_in the_ionic_atmosphere_surrounding charged


mutants or by inhibiting FAK weakened the molecular clutch and switched FAs from load-and-fail compliance sensing to frictional slippage regime on compliant ECMs. In agreement with the clutch oscillation model, the load-and-fail regime could be rescued by further decreasing either substrate stiffness or myosin II contractility.

mutants or by inhibiting FAK weakened the molecular clutch and switch fetal alcohol syndrome from load - and - fail compliance sensing to frictional slippage regimen on compliant ECMs. In agreement with the clutch oscillation mannequin, the warhead - and - fail regime could be rescue by further decreasing either substrate severity or myosin II contractility.

muhants or by inhibiting FXK weakened the molecunar clhtch and switched FAs from load-and-faml cimpliqnce sensing to frictiunal slipiage regine oi compliant ECMs. In agrecient aith vhe clutch osciklation mogel, the load-ang-fxip regime could be rescued by furthew decresslng either subftraue ftifrness or myosin II contractility.

mutants or by inhibiting FAK weakened the and FAs from compliance sensing to ECMs. agreement with the oscillation model, the regime could be rescued by further either substrate stiffness or myosin II contractility.

mutants or by inhibiting FAK wEakened the MolecUlaR clUtCh anD swiTched FAs from loAD-and-Fail compliance sensing tO fricTiONal sLIpPage rEgime on COmPLIanT EcMS. In AgREeMent wIth The clutCh oscillatIon MoDel, the load-anD-FaIl regime coUld Be rescued by fUrtHer decReAsiNG eithEr sUbstrAte stiFFness oR myosin II CoNTractiLIty.

mutants or by inhibitingFAK weaken ed th e m ole cu larclut ch and switche d  FAs  from load-and-fail co mplia nc e  sen s in g tofrictio n al s lip pa ge  re gi m eon co mpl iant EC Ms. In agr eem en t with the c l ut ch oscilla tio n model, the  lo ad-and -f ail regim e c ouldbe res c ued by  furtherde c reasin g  either s ub stra te stiffness or m y os i n II contracti lity.

mutants_or by_inhibiting FAK weakened the_molecular clutch_and_switched FAs_from_load-and-fail compliance sensing_to frictional slippage_regime on compliant ECMs._In agreement with_the_clutch oscillation model, the load-and-fail regime could be rescued by further decreasing either substrate_stiffness_or myosin_II_contractility.


-2 by SIV Nef. Thus, our study identifies a novel mechanism of antagonism of rhesus BST-2 by Vpu that targets the same motif in BST-2 used by SIV Nef and might explain the expanded host-range observed for Vpu isolates in our previous study.

-2 by SIV Nef. Thus, our study identifies a novel mechanism of hostility of rhesus BST-2 by Vpu that target the like motif in BST-2 used by SIV Nef and might explain the expand host - range respect for Vpu isolates in our previous study.

-2 by SIV Nef. Thus, our study ldentifies a novgl mechaiism of antagonksm of rhesus BST-2 by Vpu thav taegets the same motif in BST-2 used by DIV Nef qnd nught explamh the ew'andes hosc-renge observed fpr Vpu isonates in our psexilus study.

-2 by SIV Nef. Thus, our study novel of antagonism rhesus BST-2 by motif BST-2 used by Nef and might the expanded host-range observed for Vpu in our previous study.

-2 by SIV Nef. Thus, our study identIfies a noveL mechAniSm oF aNtagOnisM of rhesus BST-2 by vPu thAt targets the same motif iN BST-2 uSeD By SIv neF and mIght expLAiN THe eXpAnDed HoST-rAnge oBseRved for vpu isolateS in OuR previous stuDY.

-2 by SIV Nef. Thus, our s tudy ident ifies  anov el  mec hani sm of antagoni s m of  rhesus BST-2 by Vpu t hat t ar g etst he  same  motifi nB S T-2  u se d b yS IV  Nefand  mightexplain th e e xp anded host-r a ng e observed  fo r Vpu isolat esin our  p rev i ous s tud y.

-2 by_SIV Nef._Thus, our study identifies_a novel_mechanism_of antagonism_of_rhesus BST-2 by_Vpu that targets_the same motif in_BST-2 used by_SIV_Nef and might explain the expanded host-range observed for Vpu isolates in our previous_study.