me (613) lemfiéfl
`
`Order Ill 0508531EDP041845141
`
`Tue How 16 13:52:30 2004
`
`Page 2 of 10
`NOV 16 ’EM WNW”
`
`M0
`
`I.l
`
`ARTICiEr—mw
`
`
`Molecular Species Compositlon of the Maior Diecyl
`
`Glycemphmspholipids fmm Muesli-3. Liven Retina
`and Brain of Cod (Gadus morhua)
`
`ILV, Ball" and MI, Bleak
`NERC UM ol Amati»: Elmruamkw, Eternal of Natuml Balinese. lEnlvarnfly m Stirling. Sllrllng m0 auxW
`
`
`
`
`
`
`The mnlmulnr sum (ammunition oi emphafidylchullae
`{PC}, thphafldylathmm amine (PEl‘nml phmphafidyl»
`urine {F35 fmm 9:th muscle, liver, “tins and brain at?
`end {1326st marine} were determined by high-
`
`perfotmance liquid chm tography at the mmfive
`
`Liwtlintzylglycerul 3,5difluwanzayl derivatives“ a
`minimum“ of 69 dinnyl Knacks WM ldwtlfledl In tumult
`and liver “th fatty etidlpolyunummw fatty mini
`C?UFM and mm omtmted fatty afiWUF‘A Realm”
`species were predominant. particularly lfidlflflfi and
`lfizflifltfi in PC, 16:0!225 and 18:11'2235 in P’Eand meme
`1 d 1311322126 in PS. Edmnhnaannyl ape-lies were ma»
`
`jar mmpanenta a! PC, PE and PS- l'mm retina. mmpfla'
`ins 2‘53. ”.3 and 503% at the mpmflw whale. 1:»
`domeehemwoyl taped" were «1m abundant ln PE mud
`fee memmemmtm
`F3 hum 3mm. emulating
`
`namfiwly. DEPUF‘A enemies wm imperial,m muadel
`
`mullahs 212% in PC and 38.3% m PE. PC fro
`ail
`tissues had the lam-gun: emanate of upswing Bantams
`only animated a: monounsaturated fatty adds“ an.
`counting fur 59.3% a! PG from brain, including [2,15% a!
`10352411 elm 24:1il8;l.
`Lipids 36‘. “$573 (1901}.
`
`
`The main: ultimatum! glycm’upl'msholipida Mm vertebrate
`neural tissue contain large wants of éocuaahmaamic
`acid 1mm; 22611-3). Mwnaamrnmzl fatty acidn {PUFAE
`are very tenaciously mtaleed by neural Wee (ll. and We
`a: Ill-we geuemtlnna M chem (leprlmemm m requiem
`m Mum algm‘fiewtly the ma PUFA cement. mi brain if:
`rat {2'3 anti brain and retina. nf rhesua man keys 3]. Dept»
`
`tion mi 15-3; PUFA in brain and retina results in reduced
`
`visual and cagnitive abilities in [M33 fiankeya [4}.
`flowwer, the precise rule {of 22;6n~3 in neural blame in
`mtknm
`Early “tempta ta mmm phuaphehpidm tram Wine
`
`md WW mag-manta mam awarding to the degree of am
`
`saturation by admrption thin-1339I cinematography
`(TLCl {5}, and later by Malian TLC We showed the
`pmsance ufi fractlom containing mm than six dwhle
`bands pee molecule Mammal analyses of the fatty adder)
`mPISaml Pfilfrmmlmg mfiahawedflfisfiwambemhmh
`the 52ml md MM methane of the glymride and it; was
`deducmrl
`that; diPUFA amine were meant (7} The
`
`
`ITO when wwpmdmm ash-wild be “dreamed.
`Abbmfififima BHT‘. 11113.th hydmxymlum; GLO gwliquld
`chflmmwwhy; HPLC
`wmliq£1736 dwwm why;
`HP’I‘W Wwfmmm lhmluym Womatwaphr; MU A mu»
`nuumetumled {any amid: W3 phmphulidylehuline: PE
`phmphatitlylethanalamlne; Pl. phanphflidylfinmiml: es,
`phosphatidylsamiua; PUFA. palmumram fatty acid; E033. rod
`anter-
`ana
`t: SFA, animated {any Mid: TLC.
`thin-layer
`chxmnmgmphy; VLCPUFA, very long chain PUFA. “film
`awaits am abbnfiamd my
`follow»: «an momma PC Ea
`1~pflmlmyl»2«flwmnhenmeylvemglmfiwphmphmhwnm
`
`demlopmeat at methade fer analyzing the molecular
`species mmmeltion of glyemphmphelipida by high
`pedal-mam liquid chmmetegmphy (HPLm directly {8)
`or by pmpanng derivatives (9-12}. or by garliquld
`chromato
`hy (GL6) 01 velatile derivatives {13} has
`mauled the fillly complexity at" the lipid campamnts of
`
`memhmne #3ch andW techniquea haw: new been
`applied to may ayatem l'he pummel hf leUFA
`muleculm Wee in Ema retina wan mammal by amen
`analysis: (141E wlfile mum at? glycemphmpholipida from
`
`the brain and retina of tmut showed large amounts of
`
`
`
`di22 :5n3 apmlea, especl
`3! ill phasphatldylahaline {P03
`phasphatifiyEathanolnmlm {PE} and phmphaudylaefine
`
`(133th retire: and in PE and PK fwm brain {15L Small
`mama of MFA amine elm have been (Guild in new
`mural times including ml; srytlmzcyw {1 1}, mt mm (16},
`fish muscle (0E and and ma EH). Marine fiflh are parties):
`Early n‘cla in m3 PUFA (18$. and this paper presents
`
`molecular sped” analyses nf PG. PH and PS cf four
`tissues 01' and {Gadus marten}. The resulta from two
`neural museum. brain am We» are mmpamel with them
`limm liver and: muscle
`
`MATIBIALS AND METHOD:
`
`
`
`{Sod (Gadus mrhua} {300—600 3 weight) mm obtained
`from the Marine Station (Millport an the Firth af Clyde.
`Stanislaw”. Whammy: in u seawater aquarium at. mum
`on a diet of pred «quid axed used within We mks ul’
`capture. "Flak were killed by demplmdmn. mil the 23%.
`liver and fillet at muscle rammed and Emma at --?0°G
`until required. Emil“ m mad and uaedimmedlahely.
`Phasplnofipase C from Emilia: can!“ was obtained
`hum Boehringer Carpnmtlan mandala) Ltd. Email. East
`Stamens, England} Butyletexl hydmymluwe EBH’I‘} wee
`1?me gm: flhemlcal Cu (Wale. Datum. England! Maw
`malt whammyhumid: w Mm mmh mammal Ca
`(Gillinghm Emmet. Englamll and me Whallimd
`from cerium tetrachloride Mare use Smmmdl llpida were
`obtained fmm Sigma and {mm Nu-Chel: Prep (Elysian,
`MN}. as detailed in [15L TLC and high~petfemm thlnv
`layer chmmamwau
`(Hm:
`tea wem mated with
`mileage! 60mm armledzt. 6?le Anal»: made
`glacial acetic md. when gammmde. pmilmvv201 and
`pyridine were purchased hum BDH Ltd. {Paula Dome,
`
`
`England} AfinthumlwmufHI-‘m madamhnm
`
`humfibmlcsls mu Paeblesem Smtlandl
`01mm GUS and mtmphm uttyl H?LC mlurnma
`E26 K 046 cut 1:3 mime partials: ml arm attained fmm
`AlmNflowm (Reelmxw Imtmmwm UK Ltd High
`
`Wmmm Bum England}.
`
`“Ivanhoe and puflflmflen nfflptm All sealants. apart
`1mm those used for TLG and EPLG. contained 0.01% (ml
`
`BHT. Sampiw were etc-red at ~20“C under mltmgan be
`tween pmpamtiva proeeduma
`
`The mm was taken mm 20 end. cm WW 1% Fe
`
`ENZYMOTEC - 1019
`
`000001
`
`BEST AWEEMBLE CDPV
`
`mm m, 34:, m a {1001)
`
`AKER877ITC00073361
`
`000001
`
`

`

`Pram (513) 391-4760
`
`Order # 03038531509'04184514
`
`Tue Hm: 15 13:52:30 2004
`
`:3.
`%:
`
`slag:
`
`MM BELL ANB MI. DICK
`
`Page 3 nf 10
`NEW 1E5 WM WWWM
`
`é
`
`u I
`
`W: and mtmilrilflpmpm»%l (80:20. Wk flaw mm
`M) mhlmm £32};
`a (38 miumn was; and with
`mtthwntexfammfik-fla {fifizéam x-Mv}, flaw raw 1.2
`Within {1?}. Peaks wan: dew-Md at 254 run with a Pya
`Uniwm 4020 detector and quantified using a Shimadzu
`98,625: recording integrator {Anachem Lute“. Engiami},
`Pasha were identified 1mm mam 9f lug“, {relative rewm
`aim-z times. BET} ma the afimtiw carbon numhm‘ on the
`an“! {magician 01‘ film glymride m dmcribed by Eastman es
`mi, {8) and 'Ihkamum at at
`{3.2} using 16:053226 as a
`
`Mam-Ice peak. fiidmoaflmmwfiglycm! {Nuflhak Prep}
`
`
`
`men also avaiinbia for direct m arisen of zemfim timns.
`‘i‘ha main m3 PUFA-contnining malacular anemia were
`idmtifiud Mafiy by mm mf film My mid mathyl “mm
`{1'3}. whim mammal plus: 2020:0221 and 18:3?3421 plus:
`
`244033 had prwimaly hm awshmimd {10}. Each sam-
`Na m chromammphed Wee times in each {If the sol-
`wnt systems mind the atandan’i deviatiuna calculated.
`Where final peak was wens calmdmtad by submiun. than
`13thde dwiaflmn Of him mntributing wake! wm
`MM to aim the final Erma. Mulls am given to cum
`mama] plum fer «laxity. The leg“, {HR’I‘ x III} at all the
`malecular specim in each solvent Exam are given in the
`appendbc
`
`“mm
`
`
`
`in muscle $FMFUFA «pacing wm m mm. 1:201:11:th
`dim. comprising mar 50% «mi PG and P3 with almost.
`
`equai amounts of 15:0120:5 and 1&0fflkfi in PC and mainly
`
`18:0/22:6 in $8 {Table 1}. The 1 mt abundant.
`MUFMPUFA special in each aims and a majm“ wmpu-
`mm a! FE and P8 was lszlffififi.‘ [31“le FA specials mm
`awn abundant
`in PC t2l.fi%} and PE;
`{3843933}.
`pmiaminantly mama plus Mflflafi in WC and gamma
`in PE. Another nine diPUFA smite ware found. Only
`
`almfl amounts of alumina containing unly saturated and
`mmaunnatumted fatty adds were garment M which
`maxim was him mat abundant: mmprming 16.4% a! PC.
`The momma” armies comwmun at PC. PE and PS
`tmm "Hm was Madly similar m that of muscle {’Ihble 2L,
`$FWUFA firm the most. 31:411th warm in PC and PS.
`ami HUFAIPUFA species in PE.
`in Edi
`three
`giycmphnaphofipida 161112325 was a major mica mm»
`pasting a quarter M then PC. and Eadifiazfiflwaa abundant
`in PE and P& In “PS“. 1310f3255 was: a mum: maxim but
`
`new much 16m abunaam than is: $33 Mm mama: mamas.
`mm 18:0!22zfl wane equally abumimt: in PS from fiver. In
`PS from Liver 20:1;1‘225 was aim a major npeciw Mable
`2%. M‘Efldifinn, liver contained up w nima (HP HP); Species
`totaling 13.1% in PE with 22$th swim domi~
`amt. fipmm maintaining m1)? matummd amt mmounk
`«Emmi fatty mm mm again air’ minim: mwmm mm
`1.3mm mem 6.4% M“ m
`mum-ammmmmmm PEmd PS
`{mm retina accounting fur 393. "$1.8 and 593%, mspecu
`fiwiy mblfl 31.. Qfihfir deUFAi'Mfia m at mine“ im.
`mmm In PG. lfiflfmfi. 18mm] and lfizfif‘flfirfi mm
`1.3m only omBI‘WIfi‘W pwmm; at what than 51%.. M
`WW 3mm WM: 9.0% mm aim pmmt in PE
`and were probably very 1mg chain PUFAmnmirdng spe-
`cies £26}. In PE. lfiflflZfi, lfiilffiiifi and 139122.16 at 5-6%
`wetebaonly MMYW pmtatmomthm 2%, whilia
`in PS 13mm w” W Bhundanfi mm: 31.W cum—
`
`AKER877ITC00073362
`
`
`
`
`
`movad find tha mmninimg aw (3.3 g} hammganimd in
`10 wiumm emf nMaml‘mmfmmhanm {221* WM using a
`Palytmn 13mm difimpmn The mama {mm 4%?! frozen and
`eyes (5.? g wane} and 20 chapped {mam livem £89 5] mm:
`m):- homogmimd in 10 miumes of chlamfammFmethaml
`(2:1. HM, and: finely shipped (mun mums imam {211 g}
`17mm 20133}; wm homammd in 5 wlumw of gamma
`Mm; homgxmimtion mm lipid was mmde by Liam
`mthod of Witch at 01L {1%
`Neutral [mm was first mmmd from. the brain and liver
`
`
`
`extracts by gwparative "LC in hexanefdietlwi ethefiacetic
`acid {70:30:}, W}. v'hmphafipida were fluted from thfi
`origin with Wamfonnlmathmolfwate: (525:1, \HM‘V]. driéd
`by mmw magmatic: n and»: vacuum an MFG and finally
`undar an um 43? nitmw‘ Phuaphulimd grim mm
`”mm by TLC in. chinmfumflmthmmflwnmxm‘
`ethylamina {30:35:635, by vol} 1120} and the Hpridu detected
`under UV light after awaying with 011% (WW)
`2'.T'~dlchmmfluomacein in manhunt)! containing 0.01%
`{WW} BET. Phwphnupidfi mm «:1qu {mm We Silica gal
`“with nm Mull: wmhw a! chlflm armhmaghmaWaw
`6515:1‘ WV? and driad a9 Mam 22%inmean
`was remnwé from lipids by “traction with a ambition of
`2% (mar? HHC’DB. PC and PS were furthe: purified by
`
`TLC in cflmformhcemefnmhanoflmfic acidfwatm
`(imam! by ml} (2;), and fiemc‘tfl! and mtmchad as;
`harem. MI phmphnlipida mm finally chm rm [mutiny
`by HPTLC m mam! mmwsmpmwmiimMpmfomu
`meshanQWEWa {WM KG} iifimfizflfiflfltfi, by ml} {2‘2}.
`Lipids ware datecmd by swaying with 3% {WW} cuppa:-
`acetate in 3% {WV} phosphorfia mid and thawing at 160‘0
`far 15 min,
`
`Fatty mid mm} mm mm prepared by mWficnfiw
`
`
`in 2 ml» wf 1% QW) $0th mama Mid Sm ,
`thaw}
`at" 50°13 5m 35‘ hr under mtmgm The fatty Md camped»
`mm: of the phasphalipids was determined by GLC of fab
`
`Ly acid methyl when in 3 Mai 4336 chmmamgraph fit"
`
`ted with a G?“ Wu 5208 fused amen capillary mlumn {50
`m K {132 mm 1.11} (Chmmpmk UK. hm. Mndum
`England? i333. and with a (SP $0503 Mad 3mm capillary
`momma will
`)6; 0.34 mm id) (Quampmké may. waning
`
`hydrogen aa carrier gas,
`
`
`1%me (If flfidinitmbenmyl derimtiwa. O eimg
`
`portions of phosphnlipid were hydraiywd with
`
`phmpbnflpam C waning a ma phase: swam at 1 mL dinthyi
`9.th mi 1 ml; 0.1M amdium karate human 33“ 7.5% M:
`m tampmamm mam mmm far a In: {253, M. rm: end
`{if the imubatima 1,2«diacy1giymmln we $9:an and
`purified by TLC in hmmafliethyl a‘tmcetic acid
`Wmfiflfl, whim E17}. 1~Alkamylo2mcylglycamla and
`Mlkyl—flnmyigwwmls m separabed at this ataga
`Amman ai phwphalipid mmfining after pmpholigm
`fl: Manama mm 1w; than 2% at the: «swung mama
`mflw mm mm in dry mm mm
`M fl -V_mbamwl sauna a: we fur a min under
`nitrogen ‘exuanted and mhw‘ as timber! by 'Ihkamura
`
`fit all
`[£23, Tim pu-ity of the product was checked by
`HM“: in mmfliemyl athafimLic mid mama WW}.
`Sepamtian {if molecular apmim. The hafiimyltadiw
`tmbmzoylvmsglymmlfi WM wpmatd by HPLC at.
`19~21"‘C {m mma phase caliwmms using a 393m Unicam
`$1310 pump (Pm Unicam IML1 flambridga, England} and
`flame ham-ads aoivant system An ODS calumn was
`mad with mathmollpmpamfiwnl (95:5. W}. flaw mm M)
`
`019105. Vat 20. Na & 0W)
`
`000002
`
`
`
`000002
`
`

`

`RM: (6.13) 991w21ffifl
`
`0 flier" at 050353 1501904134514
`
`Tue Hm: 1% 13:52:30 2004
`
`Page 4 0f 10
`MW IE: ’W 03:49PM
`
`5W
`
`MOLECULAR SPECIES DI“ CflD GLYCERQPHQSPHGLIHDB
`
`TABLE? 1
`
`Mnlmsm finned“: Cammuu‘im at 93mph
`tiWMmIim. thmudylnLhAmhmfiM and
`
`Phawhafidylsuine hum Cad Muscle“
`
`Pfimmwa Pamfi%T
`
`Swain
`"WNW
`POImal‘ifi}
`MWWA
`'3."
`mm& {1 324122263
`W‘Nflifi
`W0
`202512035; 20:4}2226
`22:5!3‘226
`22:5’23fi; 22:4.‘223'5
`Tum
`
`HHHflfifi
`
`pea???Hfiwwwu
`
`6,
`
`M
`
`ryrpgwwnwmteaua
`929'9PrrH9?&fiin@wwwinQQ‘ub:
`cflfififlflfifififi
`rrewmmnbfiiwaaaF9???””W’d
`
`fiFAfiPUF‘A
`limfi
`mam5
`Mflv‘fii‘fi
`Ifimfl
`xfiww
`JHMM
`IRWJE
`mmus
`mama:
`1&W’5
`Toni
`
`WWMWWA
`WWW
`imam
`Imimw
`:mmm
`JMflM
`W'Imfi
`Total
`
`¢¢9P¢§§¢fiwwwwwmwa
`
`fl.
`
`9‘ h
`
`W
`
`1,.
`
`.‘Pmm
`
`Mflfiflfifi
`
`flag???wai—vga
`
`@f13w
`
`'6’Hflififi9?9?“Qa»-
`99Vmam-n
`
`W
`
`4a
`
`3????
`?§?§P
`
`afiuQQNG—LFQ
`9????9”gfifigfl
`
`H%HEHHRRW%
`Hflfi%fifl
`
`NW?fiwifi'
`
`Skwgwmwfidg
`
`._.fiwwwgp
`wakkawm
`
`MP
`
`“Oar:
`
`PP?OWE?
`
`&%&&ll&%&¥ 9px;
`99??”Qwwflwifib
`Hflfifififi
`
`a-o
`
`a:
`
`»
`
`B;
`
`fifipgififmmmm5ua
`{rqufifliflfiwwfifisfi
`99¢?FPFFfifP
`§?35?g?
`
`$FWUPA
`lfififilfifl
`mamas}; 11 $0120: 11
`Tom
`
`MUFANMUPA
`lfi‘ffigj
`Imam; (mummn
`"mm
`
`Very 1mg mm
`Total
`
`fifi g?WW
`
`9??man
`
`FF?W63!“ HH $9MM
`
`m4
`
`I"??? ?§1—!»
`
`P???tar-1a.
`
`3*??? 1a; a...
`
`%8?53,-«24
`
`233 3: (LG
`H
`
`Q?
`
`‘15pr in italics hiya Mmed all-1 and M4 fatty acid distributions. otherwise it was
`assumed that the: m; smturahd futy ma mm on the arm} mit- am. {July mm fatty
`acid ambinnum 5a giwn for thaw} am with mm mm”; My [mammal immers.
`mme :5de m m cmmmm baud en fatty acid mmpaailsion 453m The
`mm m giwm m ¢ 1 nmdmd demm mm m an armament denim} aim gm
`mat.)w Tim fanning 1114;112:6qu wade: wan aim dumped as, mam in at law: QM at cm:
`giywwhmpbwpida: ifizifwfi. 22:4!2815. 14133.“. 141mm imam 160220, 1811031
`‘Ifl'mfi 1&mfi Ifiiflfi Ifi'lflzfi, Rama m'lm'li Erma M1324, 151W14fl.
`24:0?! 611. IBflIZUfl, lfljflfltl. laurmi. Efidmfi, 24:1.22212 lflrlthl.
`bw. New; demcm
`cm. «A: “a.
`
`prising away EPA and MUFA totalled 31.13% 11% and 5.9%,
`mamtiwly. in FE and Pig (mm marina.
`N3: {mm and brain 5&1qu We moat; mmgilmc cornpmv
`him: M w M the glycmphmphofipids “amine-cl; a
`ufinimm {If 54 species were found, induding men
`mtknuwns which totalled 040% mm». 4}. At; least 10
`species mnmimng 24:0, 24:1 and 26:1 may acids mm}
`
`ling 153% mm foundy. (in: mm?» impnrwmt M” which W15
`24210821 plus mill?“ (123.0%. Them spasm mmtaimnfi'
`
`wry Eons: chain Emmy mid» W Maud only as Wm cam“
`pennants of PE and as [m mm 1% of 951 P3 from brain
`also contained 43.3% of spasm wutainmg SFA an: MU1w
`
`afwbkhlfififlafl(1&9%;muil&uusn1&a%}wmethfi
`
`
`must ' Immat. The only uther maim- specm present W65
`
`
`
`000003
`
`UMD$_Vm&fifirMQ.afl9m3
`
`AKER877ITC00073363
`
`000003
`
`

`

`Frans (613) 90102300
`
`QWW m DSQBERIWMMMSM Me New m 13:82:30 2004
`
`emf“ 2m
`F’aw 352
`W 16 ‘34
`fiamgpm
`
`555
`
`TMhfi 2
`
`MN. BELL AWE} SR. 010%
`
`Mwmw $9M»: Bump-dim M FMMMMWM Pmmwmmwm M
`
`vhwmmmm km W “W
`
`
` Swiss Pun-MM
`thl‘m
`mamas}
`«3&er
`mmm
`MMfi
`mamas; mammzaa
`mmm mwzzfi
`mm
`
`M a; m
`109 t 0.2
`m a: M
`J»
`
`m
`
`M x 9.1
`w x M:
`m :t on
`m a; m
`13.5
`
`m x, M
`m a; m
`m a: M
`M x 3.2
`4‘0
`
`awmmm
`ifimfi
`mmm
`mwmm
`mama
`gamma
`Iama
`mmm
`iiélmfi
`mmw
`mama
`WW5
`mmq
`“rm:
`
`MMFNPUM
`mzma
`mm“;
`Imam
`mmw
`mmm;
`mxmw
`13:15am
`I‘RMS
`mum
`m2mw
`MIMI
`"mm
`
`SIF‘NSRA
`“mm
`
`SFMM’M‘A
`mmma
`Tum
`
`MUFWMUKA
`mzmm
`mmwflmmzmw
`Tam
`
`m :2; M
`m a: m
`35.3 1 1.41
`2m x M
`m r. M
`m m: cm
`1,2 1 mm
`1.5 x M
`m a: m
`m a: 0.1
`uh
`0.1 :1: mm
`am
`
`L3 :2: w
`m :3: m
`1.5 m 9.1
`La 1 0.4.)
`a}: a: m
`m m m
`2.15 a: 21.1
`m :2: 0.1
`0.9 x m
`cm :9: M:-
`u
`
`2M
`
`M x cm
`(M a M
`125-3: m
`1m. : M
`M :2: mm
`3;} :1; M:
`M 1; m
`u a as
`m at. m
`m m m
`a}; x m
`M 1 cm
`mm
`
`m a: M
`M x 0.3
`ms : m
`u 1 a: {m
`as: a; w
`Mg m m
`{m x M
`m z m
`w 1 m
`w m m
`1M m 0.1
`4M
`
`as
`
`m:
`
`m z m
`m
`
`M :r. (m,
`1‘3 3; (m
`243
`
`M m m
`w
`
`m a; in:
`M m on
`M
`
`cm x M
`m
`m :3: M
`1M 2:: may
`M a: m
`m a; m
`0,4 x m
`1.0 :2: m1
`2m x m
`m x w
`an x m
`m x em
`mm
`
`mm
`m
`M
`w
`M: x m
`am a: M
`m x m
`M a: m
`m x m
`ma :1; M
`m :9; at:
`«.4
`
`M
`
`..
`
`M
`
`m a; m
`w
`
`2.?
`
`Very imw auxin
`ma
`M
`-—-
`mam
`WWW—muww
`
`“SW in EM“ Mm flaw md "11% m~2 tum: add anuwuum MW it was
`mm that. mm W «mum mm Mid was» an um amt mm 0m 0m MW
`ism! mmhfimflm in W}: far We spam with “in MM My Mum Wu.
`5»:me WM m mim‘mpmm Mm! am {am mid mmng mm. mm»
`mmfim mm 1 wmfimd WWW mmmmfimqmwm
`WmeMummm MWKkmswmummMm
`WWWyMW ma mamm WW um um xwm 1W
`ISMWIEW mama 1&‘11‘221‘, MIWM xxx-1mm 14:011610. 15mm 14:001th 115mm
`+ 11mm”, ISM/2&1. mama. Wiflm, mm {03112451.
`hm Hm: mm.
`”M <10“ 1 m
`
`uma m. 2a. m a {M};
`000004
`
`AKER877ITC00073364
`
` “7!,aw““inA‘"”5“"
`
`..mmmmw'mm.
`
`000004
`
`

`

`
`
`Page 6 of 10
`
`15 ’34
`323491314
`
`Order 1* 050853100P04184514
`
`Tue Nov 10 13:52:30 2004
`
`HOV
`
`. ngm (613) 991—23030
`
`
`
`
`MOLECUAH SPECIES 01' 0013 GLYCEROPHOBPHOLINDS
`
`569
`
`TABLE 3
`
`Molecular Spacin- Camponltlonn o! Phoaphntidylthollne. thphnfldyhthnmmim and
`
`thphnfidybainc from God Ration“
`
`Species
`P01010160)
`P81100051
`P31100101!
`
`diPUFA
`
`0.1 1 0.1
`00:01:10
`20.3 1 0.0
`22.0020
`21512240
`0.2 1 0.0
`Tue-1
`00.2
`
`0.4 1 00
`59.7 1 1 4
`0.4 1 0 0
`60.5
`
`1.7 1 01
`
`11.0 1 1 1
`1.0 1 01
`14.0
`
`SFNPUF'A
`10020-0
`Iflflfl'fi
`15.0214
`181W:6
`10002-0
`10020.1
`Tat-0.1
`
`MUFNPUFA
`111.205
`101.1120
`110-1004
`Total
`
`SFNSFA
`Total
`
`SFNMUFA
`16:0!13:1
`10:0!1311; (16:012031
`Total
`
`MUFAIMUFA
`18.11/33; [16:112011
`Total
`
`2.0 1 0.5
`28.2 :1: 0.1
`1.? 1 0.3
`0.0 1 0.1
`0.1 1 1.1
`0.11 1 0.0
`36.5
`
`0.6 1 0.1
`1.1 1 0.5
`0.2 1 0.0
`4.1
`
`0.1 1 0.1
`5.0 It 0.2
`0.0 1 0.0
`0.0 1 0.0
`5.0 1 0.1
`1.0 1 0.0
`14.4
`
`1 2 1 0.11
`5 0 1 0.0
`J
`
`9.0
`
`0.1 1 0.0
`1.7 :1: 0.1
`0.4 1 0.0
`1.0 1 0.5
`10.0 1 0.1
`1,0 1 0.11
`22.2
`
`0.4 1 0.1
`5.0 1 0.4
`1.1 1 0.1
`11.0
`
`0.?
`
`k“
`
`0.5
`
`13.4 t 0.1
`1.3 t 0.1
`15.3
`
`0.3 :1: 0.0
`0.5- .1: 0.0
`0.1-1
`
`0.3 :1: 0.0
`1.6 i 0 2
`2.3
`
`1.3 i; 0 1
`2.8
`
`0.6 :1: 0.0
`1.3
`
`2.6 :1: I] 1
`3.1.
`
`Unknowns
`U1
`U2
`
`5.]. d: 0. 2
`4.1 1 0.1
`
`——
`——
`
`—-
`
`— “
`
`Species in italics have rhflned "-1 and 90-2 fatty acid distributions. otharwisa it. was
`
`assumed that the mast saturated {an}! acid was on the 011—! position. Only 0114! fatty
`acid combination in given Im- thnse apncies with two equally likely positional 1001112“.
`13111011101011 spa-0102 are minor 00019011101100 based 0.11 fatty acid composition data. The
`errors are given as :I: 1 standard deviation rounded to the nearest decimal place 1000
`
`text). The following molecular 01100.03 were also detected as <09‘11: 111 at least one 01
`the glycamphnspholipids: 0002015 22:512226 22.5226. 1010220. JEWEL-5 18-00135
`18 1/20: 4,
`1&11’225 20:11:22.6 20-11225 14;0!16:0. 16:0!1810. 1810110.]. 161111611. 18:1120:1. SEMI.
`182112401.
`0—. H01. 0010:1211
`‘tr. (0.1%.
`
`
`011111101111
`1611111816. PE: and PE hum bum we
`musician to those 11:10 10119010. 11w and 104.010. D1210
`was abundant in both 11103008, Especially P8 {24.0%}, while
`
`18:0!22:fi. 13:1!2203 and 131012211 were the other major
`species in PE, and 18101226 and 13:1!22:6 in PS.
`
`DIS'SUSSION
`
`
`
`
`
`
`
`
`01111: g moat 0f the
`The method used here15 capable of
`
`
`
`
`
`molecular species present in glycerol: upholipids fm
`
`fish tissues. Ewen species containing fatty acids which
`
`
`
`small amounts ((0.5%! were not always
`mmthwy
`
`
`410th 03., 16511-8 and 181311-11. 100m of 1111501111111th
`fatty acids are not. separated. 41g. 11—? and 11-9 MU FA or
`n~3 and. n-'6 PUFA. Analysis by GLC showed that 22:5 was
`
`
`the 11-310011101“. 22: 4 the n-61110:1101- and 20:4 medorninantb'
`tha 11-6 100mm": 20:411-3 was a very minor componant in
`
`
`most samples. The 011101110.anch systems could not
`
`
`separate diPUFA 1111-00100 0011150101113 diffaent [atty acids
`
`but the same total carban number and double bonds Very
`
`long chain fatty acid-contaizfing special were 0100 difficult
`00 10001110. and the 001mm and chmmtogrinlv 001101010110
`
`
`
`
`
`
`I p 05, V01. 2.91. No. 0 (1001)
`
`000005
`
`AKER877ITC00073365
`
`000005
`
`

`

`Pram (613) 99147550
`
`0rde‘r # 05085316339041.34514
`
`Tue Hm: 16 13:52:30 2004
`
`MIN 1:5:
`
`Page 1’ 0f 10
`rem E12: 49m
`
`3
`
`2
`
`~ 53:0
`
`MN. BELL mm .m. mm:
`
`TWLE 4
`
`Mmlmim 39w“ Campmwm M Phawmufiylchuljm nghaflfiyhthumiawim and
`
`Pbowmmdflamm 1mm Cad Brain“
`
`WW
`90mm
`PEima§%:
`Pawn:
`A
`in.
`x
`M
`0mm
`20:5;22;0
`mamm:
`maxim
`mm:
`
`{1.2 i 0.0
`(1.5 ".1: M
`~91
`
`1.0
`
`M :2; {m
`13.3 :1: M
`0.2 x m
`15.5
`
`1.0 a: 0.0
`24.0 m 0.9
`m m m
`26.1
`
`spawn
`mama
`ifififlfifi
`mmw
`mmw
`IRWfi
`mmm
`mama
`macaw
`
`Tom
`
`umwnm
`1mm
`maxms
`Ifl'l/Ms-i
`Javxfia
`m'félafi‘
`Total
`
`30mm
`mmw
`152011823
`Total
`
`{SEAJM {SEA
`16mm;
`mmmm
`10:00am Momma
`Total
`
`MUFANUFA
`mum-I: museum
`magma; mama
`Mimi; mmm
`mm
`
`2.3 i 9.23
`11.5 1 0.3
`m 1 m2
`0.5 $ (2.0
`1.3 :1: 0.:
`2.2 :1: m
`0.3 1 (m
`0.9 1 m
`
`2121]
`
`cm :93: M3
`2.x :1: 1M
`0.1 .4: 0.0
`0.5 :3: 013
`11.3 :t M
`5.2
`
`1.3 a; m
`m m 0.2
`3.5
`
`1.2 :t m
`18.0 :1: M
`9.5 :t: M
`25.9
`
`8.: i (m
`3.2 :3: M
`5.3 a 0.3
`10.5
`
`1.9 a: m
`19.0 i {3.2
`1.3 g: 0.0
`m a: m
`m 1%: mm
`14.? :1: m
`1,0 :2; :11
`m 1 {1.0
`
`46.0
`
`m 3: M
`ms T}: m
`m x m
`m i m
`1.3 x 0.3
`22.2
`
`W
`0.; :2; m
`m
`
`0.2 3: 9.1
`3.1 x 9.0
`4.0 :5 M
`m
`
`m i M
`m t: m
`rm 0; m
`m
`
`m. 1: cm
`1.5 t 111
`....
`m 1: 0.1
`2.6 1: 0.13
`34.5 a: a.»
`m x 0.1
`1.; :2 0.0
`
`41.5
`
`mm a; 0.3
`m: i: m
`M t 0.1
`1.4 i 1.0
`9.5 x m
`22.1
`
`m
`.m
`
`13.:
`
`..
`0.4 :3: 0.0
`3:: :9: 0.1
`w
`
`1.? i 0.0
`m x 0.1
`3.»;
`:2: 9.0
`4.3
`
`Very 1mg chain
`iaflfiizh {2013221}
`Zi‘fiflffiéfl; lfizfimfifl
`Tami
`mam-um m
` m
`Tam)
`3.3
`m
`w
`
`12.8 i 0.2
`1.2 3: 0.1
`15.9
`
`-—
`—
`Lt
`
`llfi. i 0.1
`--
`
`(LE
`
`5f
`
`a i
`
`3
`
`‘
`5
`3.;
`»
`:
`:.
`3;
`3:;
`.
`f
`r :2
`?
`.t
`
`i
`
`:3:
`g;
`;
`:
`
`s»
`0.
`._
`«;
`
`:
`
`E
`.3:
`i
`3
`
`f.
`
`f
`
`.r
`:
`.
`
`
`
`
`
`;;
`3::
`:
`3:
`
`“39mm in italics: have dafinsd and and swfi fatty acid mummy. arbor-aim it was
`”Mind that £15: mat “Maud fatty acid was an the nun—1 amnion- Only an: Fatty
`acid mmbimu’on is given fur d'mae spedas with turn squib: iikeiy positional human.
`Brmkamd «patina we mimr amp-mama based on fatty amid mmpanifiun data. TM
`arm mm aim a :9: 1 3W dwmm macaw m m was: windmill 92m Km
`but]. Thu. {allowing anemia: Wes mm- alm {Embed M (0.9% in at 1mm; am mi um
`afimylmphoflpidm mms. mma ngfi, mama. HMS 18.0009; mmm
`”rim RUM Imam, xmm. lfiwfiih lfiflfflflfl. lfl'lflfl'fi. lfilt‘mflJflfiMfi,
`24:13am miram. mama. mmm + iifizfll‘afim. 20:02“ + mama. + Whflifl
`M'flmfi + BER/201$ “MES + ZSJM‘E
`L. Not detacLaL
`“at. 4:13.105.
`
`mm. M. 24:. Mn. 5 0091)
`
`000006
`
`‘
`
`‘
`
`"
`
`‘
`
`AKER877ITC00073366
`
`000006
`
`

`

`7
`
`firm (613) 9914750
`‘l
`
`arm i“
`
`ii 050353IWPG41MS 14
`
`Tue Nov 1E 13:52:39 rim
`
`Page 8 of 10
`NW 165; “3:1 WNW
`
`577!
`
`MQLECULAR SPECIES 0? C61) flL‘fCERflPHGEFHOLlPlDE
`
`used muld not separate: m 24:0!PUFA and 28:1!PUFA
`PM” funnel in PC from brain “this all The ma solvent.
`systems swell will) the Chi «uranium can seem“: extreme
`poaitisml immra. ass. zfizflilfizfl fmm meme but not
`16:0! 18:1 {rum 1821;16:8‘ The presumption that must
`
`saturated fatty acids are {seated at the sflwl position ef
`the glyseml maiety is. themlare. supported by this: results
`ln‘nll W3 where the method can distinguish positional
`immem.
`Mm fish 1m: parasitism rich in will HEW» Minimally
`dmosuheansin acid £183. The present melts alarm ills
`very anaemia-am minus of the molecular species of
`glycamphmphalipids item cad. PE was the most un-
`saturated glycemphosphelipicla in all times except brain,
`while PC: was the most saturated and mnmined the
`shortest average chain length. both due is a lower mm»
`partisan {if ilmfimii memes-lg apeeisel PC 12mm fish brain
`«remains a much larger menu! of SEW and MUFA»
`cantsiming species than glysarephosphslipids from other
`fish tissues 0f particular note is the large amount at
`
`13:1i24:1 plus 24:1:‘15i1, which wan found in trout. (9! and
`in swan some abundant in red. Sines cad brain contains
`1.0% @2211 family will Wilififitl may live a Mixer cumw
`mm. til“ this peek. Small manure of other swim Wilhelm
`mg my lama: chain fatty maids (24:0. 28:1} were also lewd
`in and brain PC, raising the question nf the special rule
`of these meleuular maria; in the structure and functinn
`of biomemhmnes in this tissue
`
`,
`
`The increased mphisfication of malytiesl techniques
`in (seem; we has led to them ei fili’UPA species
`m an unexpectedly large number (if tissues wall at
`erythmcyim (1 1i and tastes i16l
`fmm rat. Earlier
`molecular weeiaa analyses all fish muscle lecislflns found
`
`appreciable mums ei diPUFA species {mainly 226l22:6,
`20:5!22fl and 22:5{22y3} in slum: Enlmml {linearhyncfius
`karat.
`‘bigeyefl tuna hrazhunnw seems and Alaska
`muscle Themm ckflwgmmma. but mill. it: my (Shimmer
`couple ill); the present male :3me thaws filthy. The
`lament study elm found email amounts sf le‘UFA
`species in Pa PE and PS from cod liver and PS from mil
`
`muscle, while the large meant of di21’;6 in PE from and
`
`muscle is the greatest. pmpartien of this nelwular species
`yet mpormfi in a nanmeuml tissue
`In lamina mans Mmmmd may maids were {crawl
`in an lmi 43% 0f m 24% M: PE and 24% 05 Pt) {6}. and
`PE from fmg mains maintained: about 50% (Eli’U‘F‘A species,
`
`mainly 22:6»m3i22fim3 $14}. Tissues h-Um marine fish new
`£9.51} predamlflantly n-3 PEFH. with very little n—fl PUFA,
`and themfem diPUFA spasm in fish retina are almost m~
`timely
`emerge.
`The
`animate
`0f
`di2226
`glyumphasphollpids famed in cod retina are the highest
`email in the retina 0f any spasms anti are substantially
`
`higher than these found in New; minus (15}. Hm. we
`
`
`found that PI
`the rat-inn 0! mm (15} and cod {3?}
`does not anemia diPUl-‘A species. The melanin species
`of PI are usually dominate-ill by 13mm“ PUFA speciw
`with lameness in majm species in the retina at fish
`
`{15.27) m in iiawee 13mm mum animals @1126), but
`
`mfllwfiwa is pmduiilnmi in fish brain £15.27}. PI is
`therefore much more
`saturated than other
`
`
`glycemphmpholipids in retina.
`
`
`God are fismd feedfl‘a With the liver twin i; a major :13ng
`for storage cf neutral lipid, but Other strategies exist fur
`imaging and aiming lipiii in mm awaits 01' fish. The pat»
`
`
`
`terns of mulecular specietien of phoupholipsids in the
`titanium iii and described here therefore may not he
`mpmmfiiw at all sperms. MW labomlery swim with
`
`
`flash and ether animals have alarm; that. diam lipid
`directly inflnmm the fatty acid campaaititm of tissue
`lipids (ll. Hemm, it. has pmed wry diffimlt to deplete
`neural tiaauw all essential palmaturamd fatty acids in
`mamnmls (1-31. The presence mi large amounts of dues
`glymmphmphelipidn in the retinas at a wise range a!“
`swim species suggests a awial rule for Him minimum.
`It hm hm pmpmd that these unique lipids film! a strut:
`luml role lay interesting with rhudepain in the ROS mam-
`hmm while also maintaining at very fluid membrane in
`which conformational changes can occur during the Luv
`
`citation cycle £29}. The pigment epithelium el the we of
`many wisest fish has a reflecting layer whmh wrists of
`lipid aphemles tantalising times: exclusively iridmoaw
`hemmylglmml 63mg This unique molecule mum most
`easily be synthesized burn 3 sizes glycemphasphnlipida.
`
`Recently very long chain PUFA {“ar'I..Cl=‘Uleiilijaw with up
`in 36 carbons and six double bonds. of both the 11-6 and
`W3 wring have been found in tester [31,321 and partner
`ulnrly in PC {mm retina {28}. A sunny 13me long chain
`PUFA in refine imam a vagina}: at animal species found
`
`them to be present, 31 all systems examined, mending cod
`11%}. The latter study also fauna extrunely high levels of
`22:61:43 in cad retina and suggested a pmpom’lemnce of
`di22zfi malamflar species in this tissue (20}. as was new
`
`finned in the warmth study“ The very lung chain PUFA
`m oats-rifled w the m-I pmitim of [*0 with men-a at
`the area yaaitiem Up .33 a mini {if PC from Wine BIDS
`was found in amulet of such WGPUFN22:6 maxim [29}.
`The two unknms f: and here in PC {mm cad retina
`
`behaved in a marina! consistert with molaculw tantalu-
`ing VLJCZP‘UFA, Hm the unknowns in PC from cod
`brain were different and did nut appear be: fit this pattern“
`TM role-e at wticular fatty minis in hiammhmm strut:
`tum has been the subject a! much experimental wark and
`debate (33.341. Molecular minimum of phasphnlipld
`
`deem from fitment tissues has shown surprising
`
`specificity indieafi 3 defines: roles for some of these
`phaspholipid species in certain membranes. mer. the
`mam that make individual molecular species suited in
`particular hislaguiml
`rules. and the different firms
`amificiiim at theW mwflm in their synthesis and
`maintenance is the bismemhmm main is be elucidatml.
`
`
`
`
`mauomnamun
`
`We: are painful is Professor ML Sergei-Maud an WWW miner
`tor mmtrumw «Miriam a! the manuscript.
`
`MPEMDEGIG
`
`APPENDIX 1
`
`m resume: of Moimm Spar-ins
`Lag“, mm x m with
`
`h Mamunulfl’mflfli fill-J! WWW
`Lula
`100m meets
`lemma
`3.53:»
`Items
`31.000
`Jewell mu;
`mime mm mums
`um: mews
`1.213,
`mews
`0:137 mama
`memes
`was mama mu
`1311224
`1.2::
`News
`0.303
`lflflilim
`10m
`game , 1!
`lei/ms
`ease
`15mm 1.934
`mmme
`1.322
`{wnfinurd as most pm!
`
`“91515, Val M, Na; a (19W)
`
`
`
`000007
`
`!
`AKER877ITC00073367
`
`000007
`
`

`

`‘
`
`l
`N
`fr?!“ £01?) 00.» 2'750
`5??
`
`090M 0' 05005
`
`31650904100514
`
`Tue: MW 16 13:52:30 2004
`
`Page 9 of 10
`NOV 16 ’84!
`02:40PM
`
`
`
`2‘
`j
`
`;
`2
`‘
`
`‘
`A
`
`f
`7
`
`;
`:
`é: ‘
`3w
`2
`’"
`
`\
`;,i
`fit :
`
`i}
`“
`
`:
`5
`
`“
`
`MN. BELL MD LB. DICK
`
`1.82? APPENHLX 3
`maxim
`1.036
`Hfiflfifi
`20:10:00
`60%
`Lam
`mum};
`L036
`Mnmlazl
`mafia;
`«53.037
`10329 Lemma!” :1: m win: EmmetmlwnflMMnL-cum Hyacinth
`16:0!1810
`1.052
`mum-5
`210/326
`0.033
`
`L333 Mumnw‘fllmhmoflmh 9&5'1 WM“
`“3&6
`1.052
`IEJMZ’E
`14mm
`90852
`10m mama: mm :mnsa
`“gwfi
`1.005 mum: ma
`“"53
`“W“
`mm ‘33“
`Iawmo mm W5
`mm mamas
`1.01&
`rams
`1.235
`1mm
`mum»:
`20.02%
`0.3“
`um 13mm}:
`1"”
`1&4;sz
`mac
`“mm-a
`1.034
`16mm)
`11247
`14.02%
`”‘37 mama
`Jaemw
`ma? mma mm 2mm:
`um mmg
`mm mama:
`mats
`20mm
`1230
`ifgjifi
`2:3:
`ififij
`”3: 3m:
`:3:
`zomme mm 15211133
`1.03?
`lemma
`11m
`"
`"
`‘
`‘
`M
`'
`‘
`"
`Jam-s
`MM 22:02:25
`1.04? mm an
`1332
`”"3““
`‘3’“?
`mamas H“
`”am"
`1"525‘ ”mm was
`warm?
`1.0m mmm Lam
`gig:
`235;:
`film“
`“:33 g?“ :33;
`teams
`mm fi‘mfi
`mm maxml
`Lam
`”wig
`“m,
`$01,303
`£190
`16;a12mm
`1*813 mums
`£30,335 mmm 1.008 mama
`was
`1
`j‘
`-
`;
`31
`-
`mam»;
`6,375 mama
`1.03m
`Ara-{mg
`10403
`”M5
`W“?
`“WW “3”
`15“”2‘“
`““3 14mm
`3.377
`20mm 1.0m mmm mm
`
`mwm amt»
`MMM
`(mas
`mama
`0.040
`
`1820121531 mm “W4
`imam L130
`21mm:
`1.740 mlaw
`1mm:
`1.130
`1W”
`“3'0 mama;
`”‘3‘ mmm
`
`gamma:
`1.1135
`Emma
`my:1‘
`”mm mm 2¢Jm§s
`as“
`mm Hiram m m mama
`
`”‘3‘
`”9&3
`“3‘3
`39% Egg)
`1291mm:
`
`'
`a
`5
`“95
`
`20mm
`mamas.
`
`30:43:20.1
`5mm
`9316 mama
`
`1.1m
`1.142
`
`mmcw
`“@2215
`
`1 «ma
`1km
`1.433
`
`1.0m
`1353
`
`15“
`“firms
`QM" WWW ”42
`“3pm in mm hm Mimi m»; and W: may new 6511th mama
`1,000
`1630”“
`0300 mm”
`1‘1“
`65m». (3W cums fatty acid mkmmm Em; aim“ fur Wm swam Iflfi‘M‘S
`
`mm
`9.946 wimg
`mm mm“
`with ma muggy mubabh ”mm Emma I“ Mm it i. only Mdfim
`1:350
`ems
`:mm mm 181mm
`mink w my. mmulu a
`-‘
`“h‘
`mm x W 14.02%
`mug mama
`mm
`Lemma mm
`m” w Mam
`manage:
`min“ «firming by mm mm 0‘02.
`0.8%
`30:1 [20:11
`1.177
`24:0”. 3:31
`1.70%)
`110033051)
`03W 4
`Ifi-1f1341
`1.177
`2011;1011
`1.339
`JS‘W 5‘
`6380
`JErfl/Ifl'fi'
`1.187
`chli‘Mil
`LEM}
`15.112336
`{309% WJWJ
`12m
`923%?!
`1.839
`23:03:20.5
`1.0013
`3&‘I/HLW
`Ifigfi‘fifi‘fi
`10102
`
`
`63m in imfia have defined “-1 mad .m-R hm adnd flsuibufim
`APPENW" ‘3
`(301}! mu fatty acid mnfifimbflm in 31mm in: than: mm with km
`,
`‘
`«quail; 51W mammal Wm In wwtflm It; £0 ml); mnihla
`Mme, mm 3*: 1:3 with Mam-M m lfiflflzfil at Malawi»: 5330600 W
`
`“mm"mmfim'mn mm “”7“
`in mm: mm» mm wam Loam mm x mm value: as:
`warm!»
`«1m mmw 1.03.3
`13mm 1.3:? W by m than ME
`imam 0.0.50
`{301.23%
`1.00;
`350110;»
`1.32;
`Mariam:
`0.8453 mums
`was
`mzmaw
`3.325
`mama
`0mg
`za~1m4
`1,055
`“mm 1.243
`£63226
`0.?{11
`Jami
`1.037
`Ififl’Mfl
`30348
`gamma
`um 132mm
`1.0%
`fitmfii
`L345 “WWW“
`mama
`mm whims
`1.102
`24mg
`1.3m
`L 1mm .3. mm; Mag; mm Ram. 1:. ma»
`20:432M
`0:311
`lkflilml
`10122
`lfitliifil
`Lil-«£5
`2 Bambi}. J.M., Fiscal. 6., Durand. 6.. Mason. MM, Dam-out. 0..
`znmzm
`um 13mg
`L125 Mama
`HM
`and mama 21541080.}. Neumhcm. 43 sen-ma,
`mama's
`ohms mime
`1.130
`imam mm a. Hmmm 3%.. Wm, WEE?” m. 1mw Emmi. L.“ and km.
`HMS
`{LEW mmw 1.140 mmm 1.471
`a 09343 Pm: Nan. Amt Sm‘. USA 83,, 40214035.
`:mma
`mas
`1&Hlflzl
`mas
`momma
`laws;
`4. Hastings: m0, Comm“ WE. arm mm“ CL. and Emmi. L
`mm”
`0,533
`smfi
`1.155
`{19341.1 can Invest. 33. Eva-m
`wwwfi
`14mm
`Mama mmm ma
`0 fimmm‘gum“ LAgg'gg 510% Md 01w... 3m, ”9701.
`gamma:
`ems
`”mm 1.157 mums
`a»:
`“I"-
`“ F W M w
`‘
`‘ m
`‘
`16.133235
`10.335
`1&m4
`1.170
`29.11%!
`B. 1‘31le Mal" Md Bum. “26‘ “1983)le ilfild 35% 24. 020—623".
`
`L503
`1.53:3
`116%
`
`“$9473.
`1
`‘
`.
`.
`3. Maxim 1mm m” mi mm $1419an upmm.
`”“3
`“95 mm‘y
`mm mm,"’3
`mm";
`23 mama
`1.195 mums mm
`am mam-5
`Ifi'm£
`I‘
`‘
`-
`1.220
`18:012m 1
`1.633
`0,920
`IfiW’fi
`I$113134
`a, Nah aM.&,Hm¢.1LThk:emn. Knandhmu. K, @981 B H;
`WE 3‘2,“ m ma m madam
`‘
`’ “
`31.220
`1590000
`1.4550
`0.5330
`001%“
`“mow
`1‘1 ”WNW“: 3'” m0 Ham? LA. ”9133, J: 1w Ru. IN,
`LMS
`”3:1me
`1.?“
`mm Jfim'i‘
`IfiMM
`1w_1275_
`EBMQI
`13W
`934$
`IRWIN
`15mm um mmm um :1, Wm 59.. Blank. 0%.. MW. Rim-86} mm Beam“
`nus-m
`Imam
`16: ”30:1
`1.390
`30:1 131:1
`L913
`Mphym W 37 F379;
`9:9?“
`1 0:0” 531
`fwyllfl‘l
`1.234
`131151255}
`Lalfi
`130 WWW flu, “mm. “m ”PM H... and KW). 31L {3%51Lfmdn
`0.9M
`16:0W5
`MUIBQ
`1.296
`E31031“!
`1.9““
`fl. 359-35}-
`{3.