An Official Journal of the American Heart Association
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1 Circulatin Research OCTOBER VOL. XXXI An Official Jurnal f the American Heart Assciatin Increased Plasma Aldsterne Cncentratin in Respnse t Hemdialysis in Nephrectmized Man By Rbert E. McCa, Cnnie S. McC, Dale G. Read, Jhn D. Bwer, and Arthur C. Guytn 1972 NO. 4 ABSTRACT The nephrectmized patient maintained n intermittent hemdialysis while awaiting renal transplantatin prvides an excellent mdel fr studying the influence f factrs ther than the renal renin-angitensin system that play a rle in the regulatin f plasma aldsterne cncentratin. Peripheral plasma aldsterne cncentratins were determined using a sensitive radiimmunassay methd in nine anephric patients in the recumbent psitin immediately befre and after hemdialysis. Average patient weight fell 1.5 kg fllwing 12 hurs f dialysis n the Kiil dialyzer. Plasma sdium cncentratin decreased frm 137 ±1 (SE) t 132 ± 1 meq/ liter, plasma ptassium cncentratin decreased frm 4.8 ± 0.3 t 3.2 ±0.1 meq/liter, and the rati f sdium t ptassium increased frm 31.3 ±1.9 t 41.7 ± 0.9. Plasma aldsterne cncentratin increased frm 9.7 ± 1.6 ng/100 ml plasma befre dialysis t 17.9 ±2.1 ng/100 ml plasma after dialysis despite the lack f kidneys t prduce renin. Plasma crtisl cncentratin did nt change significantly (17.5 ±2.0 /ig/100 ml plasma t 14.8 ± 1.9 yug/100 ml plasma). These results shw that plasma aldsterne cncentratin increased in anephric patients in respnse t hemdialysis. This increase ccurred withut a cncmitant increase in plasma crtisl levels, suggesting absence f an adrencrtictrpic hrmne respnse, and was independent f the renal reninangitensin system. KEY WORDS adrenal crtex kidney crtisl angitensin renin sdium Since the bservatin in 1960 f the aldsterne-stimulating effect f exgenus angitensin in man (1, 2), cnsiderable experimental evidence has accumulated indicating that the renal renin-angitensin system Frm the Department f Physilgy and Biphysics, the Department f Bichemistry, and The Artificial Kidney Unit, University f Mississippi Schl f Medicine, Jacksn, Mississippi This investigatin was supprted by U. S. Public Health Service Grant 5 R01 HL frm the Natinal Heart and Lung Institute. Received March 27, Accepted fr publicatin August 2, radiimmunassay ptassium plays a majr rle in the regulatin f aldsterne secretin. Plasma sdium and ptassium cncentratins and pituitary secretin f adrencrtictrpic hrmne (ACTH) are thught t play imprtant secndary rles in cntrlling aldsterne secretin. Our knwledge f the regulatin f aldsterne secretin wuld be enhanced by determining whether the cncentratin f aldsterne in the plasma can be maintained within nrmal physilgical limits and whether the plasma aldsterne cncentratin can change in respnse t acute stimuli independently f OrcuUiwn Reitercb, Vl. XXXI, Octber
2 474 McCAA, McCAA, READ, BOWER, GUYTON the renal renin-angitensin system. The nephrectmized human being wh must be maintained n chrnic hemdialysis while awaiting renal transplantatin prvides an excellent mdel fr studying the influence f humral factrs ther than the renal reninangitensin system that play imprtant rles in cntrlling plasma aldsterne cncentratin. The purpse f this study was t determine the effect f hemdialysis, with its remval f electrlytes, fluids, and metablites, n the plasma aldsterne cncentratin in the nephrectmized patient. Furthermre, mdificatin f a sensitive radiimmunassay prcedure (McCaa, R. E., and McCaa, C. S., unpublished results) fr determining aldsterne cncentratin in peripheral plasma allwed us t measure this sterid hrmne with a high degree f precisin and sensitivity; the methd fr this prcedure is presented in detail in this paper. O <N CO 00 IO CO ' Q O c5 m c» c 56 S O 'O O IO "O IO O S... OCOlOtO Methds Studies in Nrmal Subjects. Six healthy vlunteer subjects were used t btain a nrmal range f values. The subjects were required t stand fr 2 hurs after having been recumbent fr at least 2 hurs. Venus bld samples were drawn befre and after the perid f standing. Five f the vlunteers were placed n a lwsdium diet fr 5 days. On the fifth day, they were again required t stand fr 2 hurs after having been recumbent fr at least 2 hurs, and venus samples were drawn immediately befre and after the perid f standing. The same five subjects were then placed n a high-sdium diet fr 5 days. On the fifth day, bld samples were drawn frm these subjects. All bld samples were cllected in tubes treated with ethylenediaminetetraacetic acid (EDTA) and immediately centrifuged. The plasma samples were frzen until they culd be analyzed fr plasma aldsterne cncentratin. Studies in Anephric Patients. Nine patients wh had undergne bilateral nephrectmies 2 t 38 mnths (average 12 mnths) befre this study were used. A summary f the clinical data is given in Table 1. Als shwn are the reasns fr nephrectmy and the length f time each patient had been nephrectmized. Each patient was restricted t 34 meq/24 hurs f dietary sdium. Other restrictins in the diet are given in Table 1. The bld pressure f all the patients was under 3 S a I 1 i TH tn ^ CO a.-2 -g a..2 S t- Q J 2 m 11* ii r-> t~- b "3 "3 t 2-2 "3 E ct S a! a a I Circtdrtin Rtsurcb, Vl. XXXI, Octbtr 1972
3 PLASMA ALDOSTERONE IN ANEPHRIC MAN 475 cntrl, and nne f the patients tk any medicatin fr 1 week prir t the study. Bld samples were drawn frm the venus side f each patient's indwelling arterivenus shunt 15 minutes befre he was cnnected t the Kiil dialyzer. Each patient was then dialyzed fr 12 hurs at a dialyzer bld flw f apprximately 500 ml/min. Immediately after hemdialysis, venus bld samples were again drawn. These tw sets f bld samples, labeled predialysis and pstdialysis, were then analyzed fr plasma sdium, ptassium, crtisl, and aldsterne. Plasma Electrlytes. Plasma sdium and ptassium determinatins were perfrmed using an Instrumentatin Labratries flame phtmeter. Plasma Crtisl Plasma crtisl determinatins were perfrmed by the cmpetitive prteinbinding prcedure reprted by Murphy (3). This prcedure is cmmnly used by many labratries and des nt need t be discussed further here. Plasma Aldsterne. The radiimmunassay methd fr determinatin f plasma aldsterne cncentratin used in this study was a mdificatin f the prcedure reprted by Mayes et al. (4). This mdified prcedure increased bth the accuracy and the sensitivity f the determinatin, and, therefre, the methd and the tests f its validity are given in detail in the subsequent paragraphs. Aldsterne carbxymethxime was prduced by the reactin f aldsterne and carbxymethxylamine hemihydrchlride (Aldrich Chemical Cmpany, Inc.) in the presence f sdium acetate and methanl. The aldsterne xime derivative was cupled t bvine albumin by the carbdiimide methd (5), and the sterid-prtein cmplex was purified by dialysis at 4 C fr 36 hurs against 20 liters f distilled water. The aldsterne xime-albumin cmplex was emulsified with cmplete Freund adjuvant cntaining Mycbacterium butyricum, and an aliqut f the emulsin was injected siibcutaneusly int ten rabbits every 2 weeks fr the first three injectins and then mnthly. Six f the rabbits develped a usable antibdy fllwing the furth injectin. Sera were stred at 10 C. An antialdsterne serum preparatin at a dilutin f 1 t 75,000 was used in this series f experiments. One-tenth ml f 8 H-l,2-d-aldsterne (10,000 cunts/min), checked weekly fr purity and isaldsterne frmatin, was added t 5-, 10-, r 20-ml plasma samples, depending n the expected aldsterne cncentratin. Samples were extracted fr 6 minutes with eight vlumes f redistilled dichlrmethane. The dichlrmethane layer was separated frm the plasma layer by centrifugatin, and the plasma layer was remved by aspiratin. The dichlrmethane extract was Circulatin R*,t*rcb. Vl. XXXI, Octbtr 1972 evaprated t dryness in a water bath at 37 C under a stream f nitrgen. Each sample was disslved in ml f a methanl-acetne (1:1) slutin and applied t Whatman n. 1 chrmatgraphy paper which had been washed cntinuusly fr 48 hurs with redistilled methanl in a mdified Sxhlet extractin apparatus (Cming Glass Wrks). The paper was remved frm the wash and dried immediately befre samples were applied. The chrmatgram was equilibrated fr 2 hurs and then was develped by descending chrmatgraphy in a benzenehexane-methanl-water (9:1:5:2.5) slvent system fr 12 hurs. Aldsterne was lcated using crtisne and crtisl standards. Aldsterne was apprximately 8 cm behind crtisne and 15 cm ahead f crtisl in this system after 12 hurs f develpment. The samples were eluted frm the paper with redistilled methanl. An aliqut was remved fr liquid scintillatin cunting t allw determinatin f percent recvery, and then tw aliquts equal in vlume t the ne measured fr recvery were evaprated t dryness in a vacuum ven at 35 C fr radiimmunassay. A standard curve was cnstructed fr each series f samples analyzed. Standards cntaining 0, 10, 25, 50, 100, 200, 500, and 1,000 pg f aldsterne (Ciba) disslved in ethanl were added t test tubes, 2,500 cunts/min f s H-aldsterne was added t each standard, and the slutins were mixed well and evaprated t dryness in a vacuum ven. In additin, standards f 100, 500, and 1,000 pg f aldsterne were added t pled plasma samples and carried thrugh the cmplete purificatin prcedure with each grup f samples extracted. Dilute antiserum was prepared immediately befre use by diluting stck antiserum with brate buffer (0.05M, ph 8.0) cntaining 0.1* gamma glbulin (human immune serum glbulin, Merck, Sharp and Dhme) and 2% methanl. Threetenths ml f the dilute antiserum was added t the samples and standards, and the slutins were thrughly mixed withut faming. The tubes were cvered with Parafilm, placed in an ice bath, and incubated in a cld rm fr 2 hurs. Cld, saturated ammnium sulfate (enzyme grade) slutin (0.3 ml) was added t each tube. The cntents were mixed well, incubated fr 15 minutes in an ice bath, and centrifuged at 2,500 g fr 15 minutes at 1 C. Three-tenths ml f the supernatant slutin was transferred t a scintillatin vial. Ten ml f phsphr slutin was added t ne vial cntaining 0.3 ml f dilute antiserumsaturated ammnium sulfate (1:1, backgrund), t three vials cntaining the amunt f 8 H- aldsterne added as a recvery standard, t
4 476 McCAA, McCAA, READ, BOWER, GUYTON three vials cntaining the amunt f 8 H-aldsterne added t the aldsterne standards, and t the vials cntaining 0.3 ml f supernatant slutin frm the tubes f the standard curve, the pled plasma samples, the unknwn samples, and the blanks. All samples were cunted in a Mark II liquid scintillatin cunter (Nuclear Chicag) with autmatic standardizatin using a 188 Ba surce. The standard curve was cnstructed by pltting the percent f unbund sterid against the amunt f unlabeled aldsterne added. The cncentratin f aldsterne in the aliquts f the unknwn plasma samples was determined directly frm the standard curve. The percent f unbund aldsterne was calculated frm the fllwing equatin. percent crss-reactivity f sterid S = (x/y) X 100. Crticsterne was the nly sterid which appreciably crss-reacted, 3.5%, with the aldsterne anrisera. Other sterids which shwed a crss-reactin were crtisl, 0.05%, crtisne, 0.05*, prgesterne, , and dexycrticsterne, 0.25%. Because f the relatively large cncentratins f the ther sterids such as crtisl in plasma in relatin t the cncentratin f aldsterne, chrmatgraphic separatin f cntaminating sterids was essential. In the paper chrmatgraphic system used, a benzene-hexanemethanl-water (9:1:5:2.5) slvent system, excellent separatin f aldsterne and crtisl was achieved with the rate f migratin f crtisl being 0.5 when aldsterne was used as a reference cmpund. The separatin f alds-,,,, cunts/min f 0.3 ml f supernatant slutin Percent unbund aldsterne =» _,. : j ^ r. 0.5 (cunts/min f recvery sample) The aldsterne cntent in the samples in nangrams per 100 miuiliters was calculated frm the frmula terne and crtisne was nt as satisfactry. Hwever, by using crtisne as a standard n each chrmatgram and nly cutting ut the area cunts/min added cunts/min recvered 100 vlume plasma used /1000, where S is the amunt f aldsterne (in picgrams) in the aliqut taken fr assay as read frm the standard curve and B is the aldsterne equivalent (in picgrams) as read frm the curve fr an equal aliqut f the water blank. Crrectin fr the mass f 3 H-aldsterne added t assayed samples was nt necessary if the cunts/min in the sample tubes equaled the cunts/min in the standard tubes. Hwever, if the cunts/min f recvery differed frm the cunts/min f the 8 H-aldsterne added t the standard tubes by mre than 100 cunts/min, crrectin had t be made fr differences in mass cntributed by s H-aldsterne. The recvery f 8 H-l,2-aldsterne in the analysis f 155 plasma samples was 72 it 2.1% (SE). Samples with a recvery f less than 50% were discarded and nt subjected t radiimmunassay. Such samples ccurred infrequently and were attributed t lsses during chrmatgraphy. Specificity f the aldsterne anrisera was determined by studying the percent crss-reactivity f ther sterids cmpared with aldsterne. Calculatins f percent crss-reactivity were carried ut as described by Abraham (6). If x = picgrams f aldsterne required t displace 50% f the 8 H-aldsterne bund t the aldsterne antisera and y = picgrams f sterid S required t displace 50% f the 8 H-aldsterne bund t the aldsterne antisera, then the well behind the crtisne t assay fr aldsterne, cntaminatin by crtisne was eliminated. T determine whether this prcedure separated aldsterne and crtisne adequately, 5 ng f aldsterne, 200 ng f crtisne, and 2 X 10 s cunts/min f a H-l,2-d-aldsterne were added t five plasma samples. The specific activity f aldsterne was determined after chrmatgraphy in the rutinely used system previusly described and after tw subsequent stages f chrmatgraphy in a tluene-ethyl acetate-methanl-water (95:5:50:50) slvent system and in an isctane-fbutanl-water (10:5:9) slvent system. Althugh the last tw systems prvided excellent separatin f aldsterne and crtisne, n significant alteratin in specific activity was bserved after the initial chrmatgraphy, indicating that adequate separatin frm crtisne was btained with nly the ne chrmatgraphic system. There was n interference frm dexycrticsterne r crticsterne, since bth migrated well ahead f aldsterne in this system and, at the end f 12 hurs f develpment, were cmpletely ff the paper. T examine further the specificity f the prcedure, the results btained by radiimmunassay were cmpared with thse btained n the same plasma samples by duble-istpe dilutin (7). As shwn in Table 2, results using the tw methds were cmparable. Fr estimatin f the blank value, samples f Circulatin Rtstarcb. Vl. XXXI, Octebtr 1972
5 PLASMA ALDOSTERONE IN ANEPHRIC MAN 477 TABLE 2 Cmparisn f Aldslerne Values Obtained by Radiimmunassay and Duble-Istpe Dilutin Duble-Istpe Subject (ng aldsterne/ 100 ml plaima) (ng aldsterne/ 100 ml puama) Subjects 1-4 were nrmal yung adult vlunteers wh were n an ad libitum sdium diet and wh remained supine at 7:00 am after at least 6 hurs f recumbency. Subjects 5-7 had taken hydrchlrthiazide (100 mg/24 hur) fr 3 days and were standing at the time the sample was taken. Subject 8 was a fur-year-ld Addisnian patient. distinguished frm zer with a certainty f P = 0.01, it was fund t be abut 25 pg/5 ml plasma r 0.5 ng/100 ml plasma. T check the accuracy f the methd, variable amunts f aldsterne were added t 5-ml aliquts f pled plasma btained frm supine subjects n a high-sdium diet. A linear relatinship was btained ver the range f 1.5 t 50 ng/100 ml plasma when the aldsterne cncentratin assayed was pltted as a functin f die amunts added. T verify rutinely the accuracy, standards f 100, 500, and 1,000 pg f aldsterne were added t previusly assayed pled plasma aliquts and assayed with each series f samples. The precisin, r repeatability, f the methd was investigated by carrying ut duplicate assays fr aldsterne n a series f plasma samples. The standard deviatin was calculated frm the frmula sr> = V (2d 2 /2N), where d is the difference between duplicates and N is the number f duplicates. The SD f a grup f 20 duplicate samples was ± 0.2 ng/100 ml plasma. distilled water equal t the vlumes f plasma rutinely used were treated as plasma samples and taken thrugh the extractin and purificatin prcesses. The mean aldsterne equivalent cntent fund in 20 samples f 5 ml f distilled water was 12 ± 3 (SE) pg/sample after crrectin fr lsses thrugh the prcedure. This blank apparently was nt a result f material extracted frm water, since increasing the vlume f water extracted did nt appreciably influence the blank value. Meticulus cleaning f the glassware, washing f the paper immediately befre use with methanl, and purificatin f the slvents kept the blank in a range that culd be tlerated. A water blank was prcessed with each series f samples. When sensitivity was calculated by determining the lwest value which culd be TABLE 3 Plasma Aldsterne Cncentratin (ng/100 ml plasma) in Nrmal Subjects n Nrmal-, Subject L.H. F.L. J.L. R.C. R.H. D.L. MEAN ± SE Sex F F M M M M Nrmal diet Supine Standing Results Studies in Nrmal Subjects. Plasma aldsterne cncentratin in respnse t pstural changes was studied in six healthy vlunteers while they were recumbent befre arising in the mrning and again after 2 hurs f ambulatin during perids f lw-sdium intake and perids f high-sdium intake. The values fr the plasma aldsterne cncentratins under these cnditins are shwn in Table 3. In the six healthy subjects n an unrestricted sdium diet, there was a furfld increase in plasma aldsterne cncentratin in respnse t changes in psture. In the supine psitin, the plasma aldsterne cncentratin averaged 5.8 ±0.57 ng/100 ml Lw-tdium diet Supine Standing IAXW-, and High-Sdium Intake Higtudlum diet S pine Standing Kmtrcb, Vl. XXXI, Octbtr 1972
6 478 McCAA, McCAA, READ, BOWER, GUYTON plasma and increased t 22.9 ±2.15 ng/100 ml plasma after 2 hurs f ambulatin. Fllwing a perid f lw-sdium intake, the plasma aldsterne cncentratin averaged 18.3 ±1.70 ng/100 ml plasma in the supine psitin and increased t 37.1 ± 2.42 ng/100 ml plasma after 2 hurs f ambulatin. Fllwing a perid f high-sdium intake, plasma aldsterne cncentratin averaged 1.9 ±0.26 ng/100 ml plasma and increased t 3.7 ±0.29 ng/100 ml plasma after 2 hurs f ambulatin. Althugh there was nly a N=9 P<0.000l pj- N=9 twfld increase in plasma aldsterne cncentratin in respnse t pstural changes fllwing perids f lw- and high-sdium intake, these values were significant in bth instances (P< 0.001). Studies in Anephric Patients. Mean values fr plasma sdium, ptassium, aldsterne, and crtisl cncentratins and fr the rati f sdium t ptassium btained fr nine nephrectmized patients immediately befre and after hemdialysis are shwn in Figures 1 and 2. When the patients were dialyzed against a dialysate with the cmpsitin f 130 meq sdium/liter and 2.0 meq ptassium/liter, there was an average patient weight lss f 1.5 kg. Nrmally, plasma sdium cncentratin changes nly slightly in respnse t hemdialysis. In this study, plasma sdium cncentratin decreased an average f 5 meq/liter frm 137 ±1 (SE) meq/liter t 132 ± 1 meq/liter (P < 0.04). Hwever, by cllecting the used dialysate frm the dialyzer and measuring the sdium cncentratin in the dialysate as well OUJ 4.0 E E CO; EUJ CO * P< N=9 N=9 N=9 P<0.04 Pre Dialysis FIGURE 1 Pst Dialysis Plasma sdium and ptassium cncentratins and the rati f sdium t ptassium in nephrectmized patients immediately befre and after 12 hurs f hemdialysis. Vertical lines indicate ±SE. N = number f patients tested. V) a E _ Q_ E ^-«EC/) 15- g> ^E ^^ c m N=9 N S P<0.02 Pre Dialysis Pst Dialysis FIGURE 2 Plasma aldsterne and crtisl cncentratins in nephrectmized patients immediately befre and after 12 hurs f hemdialysis. Vertical lines indicate ±SE. NS = nt significant, N = number f patients tested. Circulatin Research, Vl. XXXI, Octber 1972
7 PLASMA ALDOSTERONE IN ANEPHRIC MAN 479 as the sdium cncentratin in a prtin f the dialysate befre its passage thrugh the dialyzer, it was fund that an apprximate average f 450 meq f sdium was remved by the prcess f hemdialysis. Als, an apprximate average f 150 meq f ptassium was remved, resulting in a decrease f plasma ptassium frm 4.8 ± 0.3 meq/liter t 3.2 ±0.1 meq/liter (P< 0.001). The remval f sdium and ptassium frm the bdy by the prcess f hemdialysis resulted in an increase in the rati f sdium t ptassium frm 31.3 ± 1.9 t 41.7 ± 0.9 (P< 0.001). Peripheral plasma aldsterne cncentratin averaged 9.7 ± 1.6 ng/100 ml plasma in the predialysis samples and increased t 17.9 ±2.1 ng/100 ml plasma in the pstdialysis samples (P<0.02). There was n significant change in plasma crtisl cncentratin in respnse t hemdialysis, althugh crtisl cncentratin actually decreased frm 17.5 ± 2.0 /^g/100 ml plasma in the predialysis samples t 14.8 ± 1.9 ^ig/100 ml plasma in the pstdialysis samples. Discussin In the present study, plasma aldsterne cncentratin was in the upper nrmal physilgical range in nephrectmized man despite the lack f kidneys t prduce renin. Recent studies by Bayard et al. (8) als indicate that plasma aldsterne cncentratin is maintained within nrmal physilgical limits in the absence f the renal reninangitensin system. These investigatrs reprted a high psitive crrelatin between plasma aldsterne cncentratin and serum ptassium cncentratin in anephric man, and preventin f the rise in serum ptassium by the administratin f Kayexalate (sdium plystyrene sulfnate) between perids f dialysis caused the plasma aldsterne cncentratin t remain at lw levels in these patients. In the present study, we fund a twfld increase in plasma aldsterne cncentratin in respnse t hemdialysis. Weidmann et al. (9) reprted similar bservatins. Peart (10) and Mitra et al. (11) fund n alteratin in CiratUttn Rasurcb, Vl. XXXI, Octbtr 1972 plasma aldsterne cncentratin in anephric patients in respnse t gradual dehydratin induced by peritneal dialysis. Furthermre, in ur study, plasma crtisl cncentratin actually decreased during the perid f dialysis, indicating that there was n increase in secretin f ACTH t accunt fr the increase in plasma aldsterne cncentratin. Als, there was a decrease in plasma ptassium cncentratin during hemdialysis. The remval f sdium and ptassium by the prcess f dialysis resulted in an increase in the rati f sdium t ptassium. Bth the decrease in plasma ptassium and the increase in the rati f sdium t ptassium wuld nrmally be a negative stimulus fr aldsterne secretin. Apparently, therefre, the increased plasma aldsterne cncentratin in respnse t hemdialysis was nt mediated by angitensin II, ACTH, r ptassium ins. Hwever, preliminary studies in ur labratry indicate that the sensitivity f the zna glmerulsa t ACTH is increased after hemdialysis (12). A pssible cause f the increased plasma aldsterne cncentratin which we bserved in respnse t hemdialysis might be that the remval f sdium frm the bdy, resulting in decreased plasma sdium cncentratin, is a mre pwerful direct stimulus t aldsterne secretin than mst investigatrs have believed. As an alternate explanatin, the increase in plasma aldsterne cncentratin culd result frm the remval f apprximately 300 ml f bld frm the cardivascular system when the bld is diverted thrugh the Kiil dialyzer. In effect, this represents acute hemrrhage. Anther pssible explanatin culd be that hemdialysis causes increased plasma aldsterne cncentratin by decreasing the metablic clearance rate f aldsterne rather than by influencing the rate f secretin. In studies n dgs, Jhnsn et al. (13) reprted a significant decrease in hepatic bld flw with hemrrhage and sdium depletin, and bservatins by Davis et al. (14) revealed a decline in the metablic clearance rate f aldsterne. And yet anther pssible explanatin, f curse, is that there
8 480 McCAA, McCAA, READ, BOWER, GUYTON might be sme unidentified mechanism that respnds t decreased sdium r fluid vlume t cntrl the secretin f aldsterne. The significance f these data is that the nephrectmized patient can respnd t acute stimuli with increased plasma aldsterne cncentratin in the absence f the renal renin-angitensin system. Regardless f the precise mechanism(s) regulating aldsterne, these data indicate that pwerful cntrl mechanisms are still active after remval f the kidneys. Acknwledgment We wish t thank Miss Betty Prestn, Mrs. Terry Grss, Mrs. Judy Blackburn, and Mrs. Sharn Kcher fr their technical assistance in The Artificial Kidney Unit. We are als grateful t Dr. C. A. Nugent fr his advice in develpment f the radiimmunassay technique. References 1. BmN, P., Kiw, E., NOWACZYNSKI, W., BROUTLLET, J., AND GENEST, J.: Effects f intravenus infusins f valine-5-angitensin II and ther pressr agents n urinary electrlytes and crticsterids, including aldsterne. J Clin Invest 40: , LARACH, J.H., ANGERS, M., KELLY, W.G., AND LIKBERMAN, S.: Hyptensive agents and pressr substances: Effect f epinephrine, nrepinephrine, angitensin II, and thers n the secretry rate f aldsterne in man. JAMA 174: , MURPHY, B.E.P.: Sme studies f the prteinbinding f sterids and their applicatin t the rutine micr and ultra-micr measurement f varius sterids in bdy fluids by cmpetitive prtein-binding radiassay. J Clin Endcrinl Metab 27: , MAYES, D., FUHUYAMA, S., KEM, D.C., AND NUGENT, C.A.: Radiimmunassay fr plasma aldsterne. J Clin Endcrinl Metab 30: , GOODFHIEND, T.L., LEVINE, L., AND FASMAN, G.D.: Antibdies t bradykinin and angitensin: Use f carbdiimides in immunlgy. Science 144: , ABRAHAM, G.E.: Slid-base radiimmunassay f estradil-17/9. J Clin Endcrinl Metab 29: , COCHLAN, J.P., AND SCOGGINS, B.A.: Measurement f aldsterne in peripheral bld f man and sheep. J Clin Endcrinl Metab 27: , BAYAHD, F., COOKE, C.R., TILLER, D.J., BETTINS, I.Z., KOWAHSKI, A., WALKER, W.G., AND MIGEON, C.J.: Regulatin f aldsterne secretin in anephric man. J Clin Invest 50: , WEIDMANN, P., HORTON, R., FRANKLIN, S., FICHMAN, M., GRAL, T., LUPU, A., AND MAXWELL, M.: Dynamic studies f aldsterne in anephric man (abstr.). Clin Res 20:201, PEABT, W.S.: Renin and angitensin in relatin t aldsterne. Am J Clin Pathl 54: , MITRA, S., GENUTH, S.M., BERMAN, L.G., AND VEHTES, V.: Aldsterne secretin in anephric patients. N Engl J Med 286:61-64, MCCAA, R.E., READ, V.H., BOWER, J.D., MCCAA, C.S., AND GUYTON, A.C.: Adrenal crtical respnse t hemdialysis, ACTH and angitensin II in anephric man (abstr.). Circulatin 44(suppl. H): 11-67, JOHNSON, J.A., DAVIS, J.O., BAUMBER, J.S., AND SCHNEIDER, E.G.: Effects f hemrrhage and chrnic sdium depletin n hepatic clearance f renin. Am J Physil 220: , DAVIS, J.O., OLICHNEY, M.J., BROWN, T.C., AND BINNION, P.F.: Metablism f aldsterne in several experimental situatins with altered aldsterne secretin. J Clin Invest 44: , Ciradati Rtstsrcb, Vl. XXXI, Oabtr 1972
A Plasma Humoral Factor of Extrarenal Origin Causing Release of Reninlike Activity in Hypotensive Dogs
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