6 research outputs found

    Magnetoresistivity As A Probe To The Field-induced Change Of Magnetic Entropy In R Al2 Compounds (r=pr,nd,tb,dy,ho,er)

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    The heat capacity CP (T) of the ferromagnetic compounds R Al2 (R=Pr,Nd,Tb,Dy,Ho,Er) was measured at zero and applied magnetic field of 5 T in the temperature interval from 2 to 200 K. From these results are calculated the magnetic component of the entropy change, -Δ Smag (T) =S (0,T) -S (H,T). From resistivity measurements, ρ (H,T), from 2 to 300 K in the same compounds, we calculated the resistivity change due to the applied magnetic field, -Δ ρmag (T) = [ρmag (0,T) - ρmag (H,T)]. The results are compared and we observed a similar dependence between -Δ ρmag (T) and (T/ TC) m Δ Smag (T) with m=0 for T≥ TC and m=1 for T≤ TC. A simple model using a Hamiltonian considering molecular and crystalline electric fields, in a mean field approximation, is adopted for the calculus. Our results show that theory and experiment are in good agreement showing that the magnetoresistivity is a probe to the field-induced change of magnetic entropy in these compounds and can be extended to other materials. A model for the factor connecting both quantities, -Δ Smag (T) and -Δ ρmag (T), is developed. This factor contains mainly the effective exchange integral which is related to Fermi energy that in turn is related to the electron effective mass. © 2006 The American Physical Society.7413Ziman, M., (1972) Electrons and Phonons, , Oxford University Press, LondonPurwins, H.G., Leson, A., (1990) Adv. Phys., 39, p. 309. , ADPHAH 0001-8732 10.1080/00018739000101511Pecharsky, V.K., Gschneidner Jr., K.A., Pecharsky, A.O., Tishin, A.M., (2001) Phys. Rev. B, 64, p. 144406. , PRBMDO 0163-1829 10.1103/PhysRevB.64.144406Potter, H.H., (1932) Philos. Mag., Suppl., 13, p. 233. , ADPHAH 0001-8732Alexander, S., Helman, J.S., Balberg, I., (1976) Phys. Rev. B, 13, p. 304. , PLRBAQ 0556-2805 10.1103/PhysRevB.13.304Potter, H.H., (1931) Proc. R. Soc. London, Ser. a, 132, p. 560. , PRLAAZ 1364-5021Ravishankar, K., Sablik, M.J., Levy, P.M., Uffer, L.F., (1974) AIP Conf. Proc., 18, p. 923. , APCPCS 0094-243XVan Daal, H.J., Buschow, K.H.J., (1969) Solid State Commun., 7, p. 217. , SSCOA4 0038-1098Inoue, T., Sankar, S.G., Craig, R.S., Wallace, W.E., Gschneidner Jr., K.A., (1977) J. Phys. Chem. Solids, 38, p. 487. , JPCSAW 0022-3697Deenadas, C., Thompson, A.W., Craig, R.S., Wallace, W.E., (1971) J. Phys. Chem. Solids, 32, p. 1853. , JPCSAW 0022-3697Ibarra, M.R., Lee, E.W., Del Moral, A., Moze, O., (1985) Solid State Commun., 53, p. 183. , SSCOA4 0038-1098Ibarra, M.R., Moze, O., Algarabel, P.A., Arnaudas, J.I., Abell, J.S., Del Moral, A., (1988) J. Phys. C, 21, p. 2735. , JPSOAW 0022-3719Griffiths, R.B., (1969) Phys. Rev., 188, p. 942. , PHRVAO 0031-899X 10.1103/PhysRev.188.942Dekker, A.J., (1965) J. Appl. Phys., 36, p. 906. , JAPIAU 0021-8979 10.1063/1.1714260Von Ranke, P.J., Pecharsky, V.K., Gschneidner Jr., K.A., (1998) Phys. Rev. B, 58, p. 12110. , PRBMDO 0163-1829 10.1103/PhysRevB.58.12110Christen, M., (1980) Solid State Commun., 36, p. 571. , SSCOA4 0038-1098Sablik, M.J., Pureur, P., Creuzet, G., Fert, A., Levy, P.M., (1983) Phys. Rev. B, 28, p. 3890. , PRBMDO 0163-1829 10.1103/PhysRevB.28.3890Furrer, A., Purwins, H.G., (1977) Phys. Rev. B, 16, p. 2131. , PLRBAQ 0556-2805 10.1103/PhysRevB.16.2131Tsai, T.H., Sellmyer, D.J., (1979) Phys. Rev. B, 20, p. 4577. , PRBMDO 0163-1829 10.1103/PhysRevB.20.4577Milchberg, H.M., Freeman, R.R., Davey, S.C., More, R.M., (1988) Phys. Rev. Lett., 61, p. 2364. , PRLTAO 0031-9007 10.1103/PhysRevLett.61.2364Rawat, R., Das, I., (2001) J. Phys.: Condens. Matter, 13, p. 379. , JCOMEL 0953-8984 10.1088/0953-8984/13/19/104Das, I., Rawat, R., (2000) Solid State Commun., 115, p. 207. , SSCOA4 0038-1098Xiong, C.M., Sun, J.R., Chen, Y.F., Shen, B.G., Du, J., Li, Y.X., (2005) IEEE Trans. Magn., 41, p. 122. , IEMGAQ 0018-946

    Mechanisms to Housing Rights implementation in Barcelona: Balancing act between private landlords and low-income tenants

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    In recent years, Barcelona has been a pioneer in the implementation of PRO-Housing Right policies. Unfortunately, these measures can be considered too radical and not necessarily efficient in a long-term scenario. Private landlords are at the center of this challenge, being that they are the stakeholder that might potentially be the solution and negotiate a balance between the two sides of the coin. On the one hand, a housing rental real estate market is shown that depends on the owners and investors, so its stability is dictated by the interest and the ttractiveness it has. Cases such as Real Estate Investment Trusts are the financial mechanism that allows indirect investment to come to the city and higher tax collection that is reflected in the infrastructure of the city. On the other, big private landlords such as opportunistic private equity funds are responsible for managing the properties and, therefore, responsible for the conditions of the tenants. Which, in recent years, have been affected the adequacy of the house. In this way, the question that will guide this research arises, what are the challenges between real estate management models and securing housing rights for low-income tenants in Barcelona? Articles, policy documents, and annual reports will be investigated to explore the investment practices of different companies (to be defined). To establish a complete view of the local investment practices of the SOCIMIs studied. The research will be divided in three sections fallowing the logic of the research questions.Aalbers M B (2016) The Financialisation of Housing: A Political Economy Approach. New York: Routledge Aalbers, M. B. (2017) ‘The Variegated Financialization of Housing’, International Journal of Urban and Regional Research, 41, 542–554 Aalbers, M. B., Rolnik, R., & Krijnen, M. (2020). The Financialization of Housing in Capitalism’s Peripheries. Abbott, S., & McConkey, R. (2006). 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Reston, VA: American Society of Civil Engineers.Graduation laboratoryArchitecture, Urbanism and Building Sciences | Advanced Housing Desig

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Phone your order or drop a line snd our directory department solicitor will call upon you at once. Our phone number ia II*, PROrC99'ONAl ATTORNEYS^ NORMAN E. MALCOLM ATTORSKY AT LAW NOTARY I'lHUC First National Bsnk bulld'no. Palo Alto Office phon* 04. Re-rtdeei- pbon* 01 HK. WANTED. AKTNA I,IKK _ A CO UK NT INSI KA.NCE. Q. R. Hlocum, Alma Hi I.ytlon. I'h. 41 A no.*.. MM'K-sMITH, KTOVK COS* if ■ i.,,,-. general repairing. .110 High Street. I'hone tiiKHi. At liiMi.llll.i; HK •ahum; t'ltlvri-ally '.,,.;:<-. SIS unci. .1,,-,-t. I'l. • tiu llll. (iRUI'KRIKS. STATIONERY. NOTIONS. II. II. Spaoldlag. Ri'.uiMii.ir srhool block. Ph. All Wanted- Professional dressmaking snd children's sewing. MO Addison avenue Wanted — Won, ny the day washing, cleaning, ironing, sewing. cooking. Mm.A.Rodi;ti. phone ttPK EGERTON D. LAKIN ATTORNEY AT I-AW NOTARY I'llll.lC lUmoiiis llulldlng. 1"*'" ■"*-'■ flftlre plmne PalO* AJtO AAA. Rm. abeam Pnlo ARo -iotk SCHOOLS. Wanted- Dressmaking and plain sewing. Mrs. More*, 317 Alma st Illl'lt'l.l'iV sI'uitUNti (/OS i)ii.'it.ii.-iv Locksmith, F. n. -smith.'I tu (Inlr. I'hon* 010K JAPANESE OOOM I'.cture framtnjt. Tlie Nippon. 271 t'niversity ave. I'll, l.l.t.x III.ACKsMITH I . J. (*ou,m->rfonl. AM Hryant street. Plume S80Y, nmSBf HAIRY PI RK MII.K AM) CREAM II. MA I tor N. PHONE tXIMX. BeptenitH .'.1 - .1 then ele. ted tl ate in ih* ■*■:•■' i" Oakland II* ". i ■. ■ .- ■ .■ . ,i .it.ti belonged ■■■■ tl *■ Si u i ai*. ' ;i< durlut- thai time Sn...' leering Ihe nntvers'tt i >■ bai hi Id i Ilnil i"* ■ slon* ih-. Oakland water- ■ ant I - ." "tiiiiii-mit i lesi :.- ; - , Uvltles and amateoi theatrleals around the bay. He li n member ol the Phi Kappa pal HYaiornlt) Tin. position eai : I- R ■ ..' ft II tl-.-ll Couldn't Bf-*t H*- Kllanora bad l***n Mr- lloneun * i 'i m ■-. I r*ew iwitefl lo gel lil.tr: ltd I •T fill. nil.I l.-Ilo...; tU I I---I, ■*—•!> ot I,,! |..| . . BUp .■ .' - tllu-ti --ll*- ■Int nlmralh-d oil Del former is..-tr.—. ■ \i..l m >on nate .t ui:i>- - m Kllfl noral mi 'i Mi* ii"i-"n "Vaa'um. smiled ib* woman "a nl n IHllii I toy -Ami mm «l'i -l «w "Weill. «■■ > tin* rabiml v "E-tgnttti: lllllllt HUH.- -Well, yo Klliiuori, '-I Uvea n*J ■'■■■■ Tom and J--i outdone by 'i tARI'KT lT.KANI.Nt; < nniproaaed air nr eacinun. Qfbaoa, 1(17 l.jiion. Ph. IMa*. KOHAKS AMI SFPI'I.IKM Robln-Min A Ct-bbO-II. ISl '■ im. i it> atrnue. I'lione ■.-.■■ Wanted <iard-*nlng; sll kind* o' t * iirun**.! or cut, or any k'id of SOCk-; *vtntng -mil bandy7~52** C*lb- erlne *t. Phone evenings 03*>K. s-t-im Wanted-—laundry to take home. hand-Ironed table linen, embroidered doll!** etc s spe* laity.. Will go OUI hy day or hour. Mrs. Tslt. Ph. SOL S-S-tf MRS. DWIG1IT V. ROSS Teacher of Pianor~*\_^ Studio 00 Addison Av*oo**. Pteoe P- -*■ -I**0- MRS. P. O- HUNDLEY Teacher of Piano. im PareaS si*. Pboae 380Y - will The Kirobcr School ol Music CARPI*T CI.EANINi; I'a'o Alio Cnrj-vt Cleaning Works, ioi High si. i'h. ittdl- I.AIHKS' ANH OKNTI.EMEN'K TAILOR."!. Sun Tailor. N. Alh«. l-a, 000 llryiiiit *t. Phon-- .III1K - t<n«isc Wmt..,] j*. .It.! liVKl.Nt. AMI CI.KAMNf. W. II. Knlglit. BIS I nuerhlty avenue. Pbon* I7u. p \l rO MTO l.A Nl -RV First ^1 sea work. I> pt rul- lr.ll. n an 1 drilvery. Pb •a* tiV. BUCCTIUCAL WORK*-. Aifuii i:. Worry, my Ruth st. PhonO Pnlo Alto 7111. Plil HDING CONTRACTOR It. S. JEWELL -IHI High str**t. Phone .11M THE FOOTPATH. All. bare It Is. Tin* sliding mil that marki the ohl rwmember ed ipot* The ^up Ihut struck our -■ i I»iy trail Tin* crtKikftj pitth acroaa tlw lot- It left ih* road i.v *.■!!.-.u ami ciiun-b. A p*ucIU*d slmdow. uothltig raoiv, Th. : parted from tin* sliver lilreb And ended st tn* fannnouaa door No Hm* or nrnipfioi tatccd im with Oequem bends ie left w rlKtit. In- nltuif-t*. wayward curvea it put. Hut alwaya Kept tin* i1.hu In -i*:lit -Oliver Wendell Uaimee L.f« Bslts Arsn't C, Mo,i |..-..|..- ii Hsfced ... WOT :;. ,.,<| "f WOUltl ;tu-«. r. fori. " Hm il l-li t •>■■ nowml ■- *- Cork nf,- belts ar* nearly o* de the dodo Th* sobatan** nltuo** aays ummI u<iwailays is a Ohruim ;r■.". railed kapok. obUlned from i tbal grot*-, in Java Kail*-* waa to stufT niahlwsS t-r many jre tore the Idea was hit on of Ming lu | iteaiitifuii.i' buoyant, qoalltles for no j bier pur--*'*'"* Tht* ttiosi buoyani nm ' i.ri.ii Enow I**' 10 .'-t!*it**uti* is. now- I ever, until* oi i**>i»i*> beada Export ! menial biioy* have been made uf this j material, bul in»i wltb eery satisfac- , tor- nstulfM l'«i»PJ heads in*nali t"*» ! easily (n nml.,' a perfect suhetaot-**-—• Peoi-nn *• U'eeklT . ~> ■ VOU Cat: Add Xl-.ll ' BUSINESS to Vo„r PRESENT BUSINESS by JUDICIOUS ADVERTISING Classified Advr tis**nents —■ * KI.El'IP.H Al WORK llurkh-irl i: rl. Worka, -.--. Kmereon **ir**t. Phon* IH. PUREST SOFT DRINKS All Oeeocs. Palo Alto fl-xSa Work*. SIS Hlgb- Ph. BSBXs, II i:\ii i RK. I.IN'oi.EI MS, I Pllul.sTKRINt;. P. A. lurnl- tni-** Co.. '2V2 I'nh. ae, I'll. IS, STANOARIl PI IMltlNti ANH METU. W< IRKS. 1ST Hariill- ton avrnti*. I'd -in- i"i\ FOR KENT. item I'liiiii.ii.-ii lull;.!-. :i^ i freshli v, .i i tn rt and ■1 Inquire «i 001 Waverley. 7l*tf FOR RENT. Fnr iient Altr* rial! on t' ■•■ Hilrd floor of th* Simklns bu;1.1ina. Is Bvnlialilw for small m«eIlnR* at a rental of $1 60 yer tueellng En quire Ttinos ofn.-n >*or Kent Sn tm v room with, i i i.'i iii.nri.-.i couple Phone *o»K , ., j, Furnished houae of (• room* nnd '-.iti. gnrnjie reai uneble to respon- tth4e ■••.fi* Aft*'* KM Waheteei ee Idrt i i Larkln -t s.m Pranclaec S-l I llll* Pot Student* „■ or " ntrely fur nlshed rooms; ever) convenlenee: i'- i \ Klrksey, T"*'* Cowpei *t |*SO-0| FOR SALE If You Fell Out of an Airship i,..,.'.. • H'g UaiKSili Spet i.i rn '• irdfln r.. Kent Parti) fumuhod 6< ...tm hou** "-iii ii.iii. PJuuii B K ** : I '-t i'" itt-ti* Hunttj i ni Rai le unlli aod « 1 stove it heating to; Hryan I streei The Lytton apts., Lytton ni Cnwr ei -t t'li-to-dtit* t, ■■ nnd -I room ;•• .furnished Quiet, clean !»nd ■ ble t.No children,) ii-3-tf K ii item ii. t i. furnished house : room - and basement 6S0 Unl- ane ■orner Pyron street S-t7-,tm* ' "■■■ Bryai l, . ompletelj fur nlshed ungalow: r-i- ranee, water h*st*r A nvenleni < S it li s Bent Rooms, also room and Mrs C n Townsley. lit i nenue I'ln-ii* rt.'.*. 8-30 -it For Rent Furnished 7-roon* i.. u-,,- nt 607 (*h inning «■'■ Dy*be'e *■ Slo.-iun. IOI I.ytton ave I 10-tl lot Rent Furnlahed >• room hou**. |30 per month (12 Cowpei stTTFt*—Ph-oire---«#r*jrKr- 171-If W.uited Hy voting woman fttuii the es*t. position at once as maid or ast-ond WOTb M C Wanted Work for t .*r I boat dallj !>> Japanese student; «%n drtve -ind tak* rare *>f aohtu wag** Wemrm. P 0 it \ BUoford i ni.- n Ij - horn.! \\ || « ■ •• Waver let itreei MISCELLANEOUS. THR URKKNE Stanford t olVei ity. mil ripen August 22 Table - bonrd .1 specialty Ontaldn double rooms for [women. it Salratlern- Mr.*-t | I'hone Lois! .I I 8-30-Se Phone *.t:k prehch Home Laundry Btrtetly hand work. 3tsr . High street. Miss Heald's Conservatory S »*«**■ t ■ I -,11 tr* ,.;.■: •>,r-m •■ ■ ■ -■ ■ ..!--- I • • • r. r ..-■'■ Mrs. Sara Bihby-Brown TKMIIKR OK VOCIL :tl "III . Di ■ NOTARY PUBLIC. ' DICY A. fiAL'UH Notary Public 257 University Ave. For Rent—l-*urrtshed housekeep- lng to.mi-, modern: ressonable Inquire tti Kipling street. 6-Mf Conveniently furnished rooms for housekeeping: also single room* l.'-l Hamilton avenue Phone I86K fi-t-tr For Kenl In KvcrRit-t'ti Park, Unyfleld '•■ room niodern rottaoe tent iii Inquire at SSI Stanford avej Rvergreeh Park, Mnvn**id rooms, sun-hin* i -* 4 6 Hrtrollton si Kor K.-nl Fuml-di* ! n|.,n I" ei I Ihr** moms, kitihenett* nnd bath Adult*, ttn? Gllman street M-t( For Kent Furnish*<l room arate entran*-* and private I'-ttli Enquire J10 Waverley -t » :•■ tt* For Rent Fouiptately* 'urtiish-><i flat* sn.l bouse*. Appt) ***i Ucyenl street. *^ .* *.i" DRBSSUAKINQ Mrs. .Wilt* s Ooodapeed 031 ailaun *t. Palo UU Prices reuoaibta LODGES Mamanits Camp, U A of A" Wed Ate. 7:10, Fraternity HmV. F w Sherman, clerk. 4*;T Webster st Forester* of America meet Wed. *»d *erretar> . 111 Hawthorne av*nu* I iHterual Aid Assoc n meet* ..i! snd 4th Mondays, Fraternity Hall Mrs.E.C.8h*rman.sec..-t27 Webster si i DANCING MISS (AltltlK l.KWIS will reopen the children - ■laming Thnrsda) Sept |fl ■ o'clock Students' <:;i-*...~ Frida* Sept 11. -t - ., , lock Vnbei Privati Studio Tin Ctowpef *m. pi*. ,,. ti.., IIAIKDRESSING. Shampooing: Manh uilnit ttii'l H:ur Work. ('.■miniigs Made Dps MISS E CHRISTOFFERSON . Vlnimt lUUtf.. LKW I n,irt*.i) PHUNK 031, Shampooing M \t.V WO PAC1AL IHI \1MKNT II,i:; GckmIs Mulr to Order. -MRS. C MALCOLM WADE Room I. Mn.1l--i--TI--i.il*. Illork. Phone 235X CHIROPRACTIC. ~ .1. •Ill alt THORNLEY DX. , . ..[ disease spinal sdjustmenis l-ady endani Consultation free. l sWeislh Hm*! IM Cfceb lrl.rS-.A- tsi h Typewriting MRS. ARMSTRONG •'!■. i:.t ,i,r st. Pbon* S30>1 For Kent Room*-, also room and ...m! 333 Waver!*) -tr**i ndla 1100) lm-u re Times of- Tou'd probably ft—1 sorry b*for* yot reat-ti,.] Ih* .■■.:■'. Rut you'd have one «atUfai-ti"ti -.ni d know you wefl not gniog t" hu ordinary *nd Cheer Up! Tumbl* to tiii* Instead. Re origin:! la your '•■''•■ -■•■•- printing. The lr*ltn kind diM-11't cost any more We fur nlsb Ideas s* well a* execute youi work speedily and right up to thr mla Bt* ■ i 1 .. Six per *ent A. A. Rai :••, KINDERGARTEN. 1 Mi*- Rvelyn ; len it im open A ■ I i Fall For Us! state highway. Mayfield. on the 3-17-lm Four Free Deliveries Daily 8 A. to. 11 a. m. 2 p. to. 5 p. M. Our Phor.t Is 3S0K CAMPUS Bvery Day At 3 P. ,*-1. Get Our Prices The Package Store E. W . Cooke Proprietor HOME MADE CAKES:-- W* l*-ei* to annoumc to tii* irml* 11 ■,-,• arTansements lone been nuute with Mm* Spvueee, trim lun put.tm.,^1 \|tv RCnarsTs .uke .ind |MMftri baslDees, to sappt* ua unl, |lf- dollclowi (hoitNMUade cake*. A K,„wi n-M.nn.ent wi||*ht on band Tlmrs- (Sny afternoon, Augu-t •Jim., an-] thereefter win i-,. able 10 supply your want*. Special order* tsi'l r*ce*lve our careful ruiroilon. EARLE & CO. GROCERS Masonic Temple Bldg. Phones 837 and 83

    Delta modulation techniques for low bit-rate digital speech encoding

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    Includes bibliography.Two new hybrid companding delta modulators for speech encoding are presented here. These modulators differ from the Hybrid Companding Delta Modulator (HCDM) proposed by Un et al in that the two new encoders employ Song Voice Adaptation as the basis of the instantaneous compandor, rather than Constant Factor adaptation. A detailed analysis of the performance, both objective and subjective, of these hybrid codecs has been carried out. Results show that overall the two codecs developed as part of this project are better than the HCDM codec. In addition the new codecs offer simpler implementation in digital hardware than the HCDM. A Computer Aided Test (CAT) system has been developed to simplify the design and test processes for speech codecs

    Projection of acoustic fields using the Fourier transform

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    A method is presented for projecting acoustic fields using the Fourier transform. It is shown that the source velocity distribution can be represented by a number of sinusoids. Each sinusoid gives rise to a plane wave whose direction is given by the wavelength of the sinusoid. The plane waves are summed at the plane of interest to obtain the resulting pressure field. Errors are introduced when projecting pressure fields by large distances. These are circumvented by observing that the farfield pressure pattern can be found by simply taking the Fourier transform of the nearfield pressure distribution. A rectangular source is simulated on the computer; the Fourier transform technique of projecting fields is compared to a direct integration technique. The Fourier transform technique is used to back project a measured pressure pattern to detect defects on the transducer face. Measurements of pressure are made in the nearfield of a circular transducer. These measurements are forward and back projected to give the pressure and velocity at other planes

    Characterisation of novel matrix-binding interactions for latent transforming growth factor-β-binding protein-2 (LTBP-2), with emphasis on heparin and heparan sulphate proteoglycans.

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    Elastic fibres are important components of the extracellular matrices, being composed of an elastin core and fibrillin-microfibrils around the periphery. Elastic fibre formation is a complex developmentally regulated process whereby fibrillin-microfibrils act as templates for the deposition of elastin. Additional matrix macromolecules, including fibulin-4, fibulin-5 and as yet unidentified heparan sulphate proteoglycans (HSPGs), have also been identified as playing important roles in this process. Fibrillins-1, -2, -3 and latent transforming growth factor-β binding protein (LTBP)-1, -2, -3, -4, associated components of fibrillin-microfibrils, make up a superfamily of extracellular matrix proteins. Fibrillins and LTBPs share a high degree of structural similarity since they both have rod-like structures of tandem EGF-like 6-cysteine repeats interspersed with unique 8-cysteine motifs. LTBP-1, -3 and -4 covalently bind TGF-β and target and store the latent growth factor in the matrix. Unlike the other LTBPs, LTBP-2 does not bind latent TGF-β and its function is poorly understood. LTBP-2 has been shown to bind fibulin-5, an elastin-binding protein and through this interaction it may target tropoelastin-fibulin-5 complexes on to fibrillin-1-microfibrils during elastic fibre assembly. In order to understand more about the role of LTBP-2 in the assembly of elastic fibres and to identify other novel functions, this study involved screening for potential molecular interactions of LTBP-2 with other matrix components, particularly heparin/HSPGs. In elastic tissues HSPGs are found on cell surfaces as syndecans and glypicans and in basement membranes as perlecan. Full length human recombinant LTBP-2 (rLTBP-2) was expressed in 293 EBNA cells using a modified pCEP-4 vector and purified by nickel affinity chromatography. Upon validation of the purified protein using western blots, solid phase binding assays were used to screen for interaction between rLTBP-2 and heparin. Heparin serves as a useful model for heparan sulphate; due to the lack of adherence of heparin to microtitre plates heparin-BSA conjugate was synthesised and purified for the binding assays. Recombinant LTBP-2 was found to interact with heparin-BSA conjugates using an established solid phase binding assay. The binding was blocked by the addition of heparin (but not chondroitin sulphate) to the liquid phase, confirming the specificity of the interaction. Furthermore, the binding was blocked by the addition of 5mM EDTA and 5mM EGTA, showing that the interaction was cation (calcium) dependent. An apparent Kd of 14.5±3.7nM was calculated from non-linear regression analysis of the LTBP-2-heparin binding curve, indicating a strong affinity. To identify the location of the heparin binding site(s) on LTBP-2, expression constructs were produced encoding three fragments of LTBP-2, i.e. rLTBP-2NT(H), rLTBP-2C(H) and rLTBP-2CT(H), corresponding to the N-terminal, central and C-terminal regions of the molecule. Good yields of rLTBP-2C(H) were obtained using the pCEP4-293-EBNA system, and rLTBP-2CT(H) was previously expressed and purified by members of the Gibson laboratory. However, difficulties were encountered with the rLTBP-2NT(H) expression construct and no LTBP-2NT(H) was available during the candidature. The central fragment LTBP-2C(H), (but not the C-terminal fragment LTBP-2CT(H)), was found to bind heparin. However, the apparent Kd of 52.2±6.9nM was significantly higher than that for full length LTBP-2, indicating that LTBP-2C(H) had relatively lower heparin-binding affinity. This result suggested that an additional heparin binding site(s) is present in the N-terminal region of the molecule. It was considered that the true tissue ligand(s) for LTBP-2 would be a HSPG rather than heparin. Therefore, LTBP-2 was screened for interaction with HSPGs, recombinant syndecans-2 and -4, and endothelial cell-derived perlecan. Interestingly, LTBP-2 bound strongly to r-syndecan-4 but not r-syndecan-2 even though both molecules were produced in the same mammalian cell system and had been screened for binding to HS-binding growth factor, fibroblast growth factor-2. This finding indicates that LTBP-2 does not interact with all HS and must recognise specific microstructures within the heparan sulphate chains. It appears that syndecan-4 is now a strong candidate as mediator of LTBP-2-cell signalling. LTBP-2 was also found to specifically interact with perlecan in a cation-dependent, heparininhibitable manner. Confocal immunohistochemical studies using foetal human aorta showed that LTBP-2 and perlecan generally had distinct distribution patterns within the medial layer, located on fibrillin-microfibrils and basement membranes respectively. However, there were small but widespread regions of LTBP-2-perlecan colocalisation which showed a similar pattern to the fibrillin-1-perlecan colocalisation. Thus it would appear that LTBP-2 is present at microfibril-basement membrane interfaces and may be involved in stabilising the interaction between these two structural elements of the matrix. This concept needs to be confirmed at the ultrastructural level. The interaction of LTBP-2 with perlecan during embryonic development is also worthy of investigation. In parallel studies, a proteomic approach was used to identify other matrix binding proteins for LTBP-2. This was carried out in parallel with another matrix protein of poorly defined function, transforming growth factor-beta-inducible gene-h3 (βig-h3). Recombinant LTBP-2 and βig-h3 coupled to sepharose were used as bait proteins to screen complex mixtures of matrix proteins from the elastic tissue nuchal ligament and basement membrane preparation Matrigel. Initial studies showed that non-specific background binding to these proteins was a major problem. In efforts to overcome this difficulty, binding conditions were varied with limited success. A two-dimensional gel approach was used to fractionate and compare proteins binding to LTBP-2 with those binding to βig-h3. The differentially displayed protein spots were to be identified by mass spectrometry. However, complications in comparing the patterns on two separate gels made identification of candidate spots challenging. Finally, CyDye DIGE fluor dyes were used to fractionate proteins binding to LTBP-2 and βig-h3 on the same gel, but unfortunately this work could not be completed in the time frame of the candidature.Thesis (Ph.D.) -- University of Adelaide, School of Medical Sciences School, 201
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