THE UNIVERSITY OF MANITOBA STUDIES ON OVINE PLACINTAL ...

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t34 FRACfTON No. (l.5mt p6r tubo¡ Figure ì0. Gel filtration on Sephadex G-b0 column ( I.4 X 94 cm ) of the fractlons contalnrng opl fro¡¡ DE-sephadex A-2s corumn (fractions #20-30). The column was equiübrated with 0.rM anmoni um bÍcarbonate, pH 8.2 .
TABLE I TABLE OF PURIFICATION OF OVINE PLACENTAL LACTOGEN PURIFICATION FACTOR I j * PROCEDURE PROTEIN RRA-GH RIA-oPL REC0VERY IN Z OF (s) (ms) (ms) pR0TEIN RRA:cH RrA-opL Extraction 646 390 3.l2 .l00 l0O 100 Acidification 546 370 294 g5 95 94 (pH 6.s) 2 (¡ 3 DEAE-Cellulose 150 ]53 120 23 39 38 Acidification 103 134 .l14 16 34 37 (pH 6.0) 34 CM-Cel l ulose 3,2 67 52 O.S 17 17 86 CM-Sephadex 0.65 32.2 27.1 0.10 8.3 8.7 ( c-5b) 446 Sephadex G-100 0.103 23.8 2Z.Z 0.016 6.1 7.1 1685 DE-Sephadex 0.021 '15.6 '17.ì 0.0033 4.0 5.s ( A-25 ) Sephadex G-S0 O.OIS** ß.t 16.3 0.0023 3.8 5.2 * Protein measured by Lowry method ** Dry welght N.B. This procedure was empioyed for the purificatìon of oPL uslng 3 separate batches of pìacentaS yielding ' a total of 35 mg oPL. In general , the results were simllar to the data shown above.
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THE UNIVERSITY OF MANITOBA STUDIES
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To My parents and wife: Mn & Mrs. K
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CONTTNT S I. REVIEW 0N PLACENTAL PR
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V. RESULTS E. STUDIES ON THT BIOSYN
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Peptide Hormones: ACTH CG FSH GH LH
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LIST OF FIGURES FIGURE PAGE I Linea
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22 Radioreceptor assay for oPRL us.
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Effect of PGF,. and lactogenic horm
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ABSTRACT STUDIES 0N 0VINE PLACENTAL
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eceptor-b i nd i ng studies show th
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-2- years have seen an increasing i
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-4- secretion of a peptide hor:none
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NHt-Aì a-Pro-Asp-Val -Gl n-Asp-Cys
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6a NH, -Ser-Lys -G'l u.Pro -Leu -Ar
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-8- 197J-; Rayford et al | !9721 .
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-10- sialic acid díd not alter the
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-L2- and Biasucci (1969) denonstrat
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_14_ addition of CAMP (Handwerger e
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-16- conponents I the.faster haying
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_18_ by hcc, as evidenced by increa
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_20- (Mason et a1 , L973i Menon ¡
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21a NH, -Vaì -G1 n-Thr-Val -Pro-Le
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-23- growth-promoting ef,fect, wher
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-25- diffusion teÊt. Josi¡novich
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-27- v¡hereas in the tissue extrac
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-29- found products of molecular we
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-3r- vÍii) S.ecreLion and metabol
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- 33 - metabolic factors may influe
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-35- Although hpI, si¡nuLates the
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-37- C. CIIHER ITUMAN PLACENTAL PEP
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_39_ stimulating activityf is found
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-4t-- (iii) Human Chorionic Cortlco
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- 43 - syncytiotrophoblast (Tatarin
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- 45 - Ib SUBPRTMATE PLACENTAT PR.O
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-47 - samples, signif,icant elevati
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-49- required for the chorignÍc ao
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B. (i) -51 - MONI(ET PLACENTAL I,AC
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- 53 - brith 13 minutes for hpl,. T
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55 Pregnarit Marê Sêrum Gonâdót
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II b. NON - pRl¡late PT,ACENTAT.
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(iv¡ Inmuhologicat Mêthogs: - 59-
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_ 6t_ 1974; Fellows et a1, 1974¡ H
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63 Ordêr Rodêhtia (Tat, nicê, ro
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Guinea Pi g: 65 Hypophysectomy of t
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-67 Orilet Perissodactyla (Horse an
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-69 activity in the N-acetyl-lactos
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(Íi) coat: -71 The goat is a spont
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- 73- When serum from a pregnant go
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-75 Gusdon et al (1920)using . haer
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77 (c) Imnunologicaì Properties of
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-79 - SECTION II : OYINE REpROpucTI
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BI which contain' as lvell as the o
Page 105 and 106: 83 ce11s usually contaln either num
Page 107 and 108: 85 non-pregnant values did not diff
Page 109 and 110: 8rl Leaf blank to corect nr.rmberin
Page 111 and 112: - 89- luteolysis when supesistpessd
Page 113 and 114: 9t SECTION IV¡ MtTttODs AND IIATER
Page 115 and 116: - 93- (.TCA) solution were added co
Page 117 and 118: -95 - (ÞPRL' NIH-P-B3) , and all q
Page 119 and 120: 97 buffer, the eluates were pooled,
Page 121 and 122: Char4ctêf ization Pfocedures l. Ch
Page 123 and 124: t0t (c) À.nalvticaL .{rod.iu¡n do
Page 125 and 126: 103 B : MFTÍÍOD OF DETECTION AND
Page 127 and 128: AFsa)¡- ProceiiureS 105 Specific b
Page 129 and 130: 107 ', C : IIEBI{OÐ OF DEVELOPMENT
Page 131 and 132: 109 D. RTNG PREGNôXCY (l) Maternâ
Page 133 and 134: l1ì (wtlvo'). After the hornogenat
Page 135 and 136: (4) Tissúe distributiori df radloi
Page 137 and 138: l'1 5 (2) Procg.ssjnq, gf. incuÞ¡
Page 139 and 140: 1.l7 for 30 min. Then the precipita
Page 141 and 142: (2) Assav of ìuteo.trgpic effec.t
Page 143 and 144: EXPERIMENT # TI (7 days after HCG)
Page 145 and 146: 123 to progesterone (l/300 djlution
Page 147 and 148: SECTION V : RESULTS (r) LACTOGEN DU
Page 149 and 150: 127 Fractioñâtión of oPL. from c
Page 151 and 152: 129 Figure 6. carborymethyì(cM,)-c
Page 153 and 154: t 3't óo0 ,oo 800 900 FRACIION r,t
Page 155: E o ø ô¡ t- z ao É, o '133 .--.
Page 159 and 160: 137 Figure lì. A, Poìyacrylamide
Page 161 and 162: Leaf blank to correct nr.rmbering
Page 163 and 164: 14r Anal.vtical Sodi um Dodecyl Sql
Page 165 and 166: o 9 x F- -)- 9 rU I æ -t f () LT'
Page 167 and 168: ¿ o s : t45 Figure 15b. Displaceme
Page 169 and 170: HoR MoNE coNc€NrRArto ln ß/mt )
Page 171 and 172: Tissue used for binding ôtudies Li
Page 173 and 174: Noñ. p..s'ro¡r {N,p.l Pr.eñonr (
Page 175 and 176: ¡( É T 2, \ ¡a.2¡./ir ,..r.¡.
Page 177 and 178: P 80 \ x.. ¡,azrot¡j .1.¿¡i¡ .
Page 179 and 180: o c o U o òs -o c2o ao eÀo o 'o .
Page 181 and 182: F 2 o o o\ ¡¡ig ô 2 Þ 0 (¡ l5e
Page 183 and 184: ! o 't6 l HORMO¡¡ E COr{CENr FAIt
Page 185 and 186: 163 Figure 24 shows the sensltivity
Page 187 and 188: 165 D. STUDIES ON THE SECRETION OF
Page 189 and 190: E .Dc À o - art & 167 Flgure 26. P
Page 191 and 192: lJhen blood samples were taken four
Page 193 and 194: Sheep no. l{6 25í 960 464 5?0 680
Page 195 and 196: 't73 fetal cotylêdons at various d
Page 197 and 198: TABLE VI OPI CONCENTRATTON IN FETAL
Page 199 and 200: l. Súrglcat Method 't77 Figure 3l
Page 201 and 202: = >€ ,3 ; I 179 rwt 76-0450 o----
Page 203 and 204: dUrlng PrêOnancy in thè Ewes tBr
Page 205 and 206: ! ñ' l83 NoRMON! C oNCENTßArlO þ
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l ol
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I I I "+ _t rl \l :- 'i rl -l l87 a
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l89 Flgure 37. Percentage ot l2sl-o
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19t column is shown in Figure 3BA.
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- 193 3H -couNdro _________SPECtFtC
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TABLE IX /- BIOASSAY OF qROIItIfH p
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z o 197 Figure 40, Radioreceptor as
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5A L lNl Pgtzq cô-tlzt Pgfl6 + cl
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SFCT.IoN II : pl.scussr0N 4d ã 1È
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Ch aracteri zati on 203 Analytical
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205 size, corresponding to the mole
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207 the specific binding of oPL to
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209 than with prolactin-like effect
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2t1 the jncrease in oPRL binding si
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213 The resuìts obtained indicate
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215 on the role af PL in the fetus.
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Surgi ca1 method- 217 Our studies o
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219 5.Relationships amonq opRL. oGH
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221 The nature of thelarge nolecula
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223 data argue for a close interact
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]. TABLE OF CHEMICAL PROPERTIES GUI
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cor,t coAT (bPL) (cpl) 3. TABLE OF
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À) E 229 Figure 43. The secretion
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231 H. POSSIBLE ROLT OF OVINE PLACE
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233 Our studies on the blnding of o
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235 through its binding to cotpus l
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237 Furthermore, oPL neasurement in
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239 circulationand alìantoic f'lul
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SECTICN IX PUBLICATIONS A) Manuscr
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SECTION X: BIBLIOGRAPHY Abramovich,
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Beas,F., Saìinas, 4., GonzalezrF.,
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247 Brody,S,, and Carlstrom,G.,J.Cl
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249 Crosignanì ,P.G. , Nencioni ,
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25t Friesen,H.G., Endocrinoìogy 76
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253 Grinwich,D.L., M. Hichens, and
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25s Hum, V.G., Knipfel , J.G., and
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?57 Knopp,R,H,, H,J. Ruder,E. Herra
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259 Manku,M.S., and D.F. Horrobin,
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261 , :iir .,.-;;r-,:, Morgan, F.J.
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263 Pierce,G.B., Midþley,A.R. and
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?65 Shepard, T.H.., Gen. Comp. Endo
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267 Tsuruhara,T., Dufau,M.L., Hickm
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