Nephritic Factor of the Classical Pathway of Complement: IMMUNOGLOBULIN G AUTOANTIBODY DIRECTED AGAINST THE CLASSICAL PATHWAY C3 CONVERTASE ENZYME

• Text
• PDF

Abstract

A factor, functionally characterized by its capacity to stabilize the normally labile classical pathway C3-converting complex of the classical pathway of complement, has been isolated from the serum of one patient with a case of acute glomerulonephritis, subsequent to a cutaneous infection. The factor confers long-lived stabilization of classical pathway C3 convertase complexes formed both in the solid (sensitized sheep erythrocytes bearing activated C̄1̄ and the classical pathway C3 convertase) and fluid phase. The half-life of such stabilized C3-cleaving enzymes extended beyond several hours at 37°C. The stabilizing activity was associated with a protein fraction immunochemically identified as immunoglobulin (Ig)G, a sizeable population of which exhibited a gamma chain of 60,000 daltons. The IgG-associated stabilizing activity was found to bind to the classical pathway C3 convertase enzyme via a fragment bearing the antigen-binding site of the molecule [F(ab)2 and F(ab)]. Such binding was demonstrable for classical pathway and not for alternative pathway C3 convertase. Thus, the stabilizing factor behaves like an autoantibody to the C3-converting complex of the classical pathway of complement. The binding of the antibody to the enzyme affords protection of the latter against decay-degradation. By analogy with the nephritic factor of the alternative pathway situation where IgG autoantibodies specifically bind to alternative pathway C3 convertase enzymes and protect them from degradation, the functionally unusual IgG in our patient was designated as the nephritic factor of the classical pathway. Indirect evidence suggests that nephritic factor of the classical pathway-IgG might be of the IgG3 subclass.

Authors

L. Halbwachs, M. Leveillé, Ph. Lesavre, S. Wattel, J. Leibowitch

Captopril-induced Changes in Prostaglandin Production: RELATIONSHIP TO VASCULAR RESPONSES IN NORMAL MAN

• Text
• PDF

Abstract

Captopril is a potent hypotensive agent whose efficacy has hitherto been attributed to its ability to alter either angiotensin II formation or kinin degradation. Our purpose was to examine captopril's acute effect on prostaglandin production, because changes in neither the renin-angiotensin nor the kallikrein-kinin systems appear adequate to account for the fall in arterial pressure. The plasma levels of angiotensin II, kinins, and prostaglandins were determined in response to increasing doses (5, 12.5, and 25 mg) of captopril and these responses were compared with the change in arterial pressure observed in nine supine normal male subjects studied on both a high (200 meq) and low (10 meq) sodium intake.

Authors

Stephen L. Swartz, Gordon H. Williams, Norman K. Hollenberg, Lawrence Levine, Robert G. Dluhy, Thomas J. Moore

Eosinopenia of Acute Infection: PRODUCTION OF EOSINOPENIA BY CHEMOTACTIC FACTORS OF ACUTE INFLAMMATION

• Text
• PDF

Abstract

One distinctive aspect of the response to acute inflammation involves a rapid and persistent decrease in the numbers of circulating eosinophils, yet the mechanisms of this eosinopenia are undefined. One possibility is that the abrupt eosinopenia may be the result of release of small amounts of the chemotactic factors of acute inflammation into the circulation. These studies were designed to examine the numbers of circulating eosinophils after an intravenous injection of zymosan-activated serum, partially purified C5a or the synthetic peptide, N-formyl-methionyl-leucyl-phenylalanine. Each of these factors caused a virtual disappearance of circulating eosinophils within 1 min, a transient return of eosinophils to ∼50% of control levels after 10-90 min, and a subsequent decrease which persisted for 5 h. In contrast, the numbers of circulating heterophils, although dropping transiently, rapidly returned and rose to elevated levels for 6 h after injection. The response was not caused by adrenal mediation as it occurred normally in adrenalectomized rabbits. Two chemotaxins of allergic inflammation, histamine and the tetrapeptide valine-glycine-serine-glutamic acid, did not cause significant eosinopenia.

Authors

David A. Bass, Thomas A. Gonwa, Pamela Szejda, M. Susan Cousart, Lawrence R. DeChatelet, Charles E. McCall

Mechanisms of insulin resistance in human obesity: evidence for receptor and postreceptor defects.

• Text
• PDF

Abstract

To assess the mechanisms of the insulin resistance in human obesity, we have determined, using a modification of the euglycemic glucose clamp technique, the shape of the in vivo insulin-glucose disposal dose-response curves in 7 control and 13 obese human subjects. Each subject had at least three euglycemic studies performed at insulin infusion rates of 15, 40, 120, 240, or 1,200 mU/M2/min. The glucose disposal rate was decreased in all obese subjects compared with controls (101 +/- 16 vs. 186 +/- 16 mg/M2/min) during the 40 mU/M2/min insulin infusion. The mean dose-response curve for the obese subjects was displaced to the right, i.e., the half-maximally effective insulin concentration was 270 +/- 27 microU/ml for the obese compared with 130 +/- 10 microU/ml for controls. In nine of the obese subjects, the dose-response curves were shifted to the right, and maximal glucose disposal rates (at a maximally effective insulin concentration) were markedly decreased, indicating both a receptor and a postreceptor defect. On the other hand, four obese patients had right-shifted dose-response curves but reached normal maximal glucose disposal rates, consistent with decreased insulin receptors as the only abnormality. When the individual data were analyzed, it was found that the lease hyperinsulinemic, least insulin-resistant patients displayed only the receptor defect, whereas those with the greatest hyperinsulinemia exhibited the largest post-receptor defect, suggesting a continuous spectrum of defects as one advances from mild to severe insulin resistance. When insulin's ability to suppress hepatic glucose output was assessed, hyperinsulinemia produced total suppresssion in all subjects. The dose-response curve for the obese subjects was shifted to the right, indicating a defect in insulin receptors. Insulin binding to isolated adipocytes obtained from the obese subjects was decreased, and a highly significant inverse linear relationship was demonstrated between insulin binding and the serum insulin concentration required for halfmaximal stimulation of glucose disposal. In conclusion: (a) decreased cellular insulin receptors contribute to the insulin resistance associated with human obesity in all subjects; (b) in the least hyperinsulinemic, insulin-resistant patients, decreased insulin receptors are the sole defect, whereas in the more hyperinsulinemic, insulin-resistant patients, the insulin resistance is the result of a combination of receptor and postreceptor abnormalities; (c) all obese patients were insensitive to insulin's suppressive effects on hepatic glucose output; this was entirely the result of decreased insulin receptors; no postreceptor defect in this insulin effect was demonstrated.

Authors

O G Kolterman, J Insel, M Saekow, J M Olefsky

Leucine Oxidation and Protein Turnover in Clofibrate-induced Muscle Protein Degradation in Rats

• Text
• PDF

Abstract

Treatment of hyperlipidemia with clofibrate may result in development of a muscular syndrome. Our previous investigation (1979. J. Clin. Invest.64: 405.) showed that chronic administration of clofibrate to rats causes myotonia and decreases glucose and fatty acid oxidation and total protein of skeletal muscle. In the present experiments we have investigated amino acid and protein metabolism in these rats. Clofibrate administration decreased the concentration of all three branched-chain amino acids without affecting those of others in muscle. Studies to examine the mechanism of decreases in muscle concentrations of branched-chain amino acids showed the following: (a) Plasma concentration of leucine was decreased, whereas there was no significant change in the concentration of isoleucine and valine. (b) Liver concentrations of all three branched-chain amino acids remained unaltered. (c) The uptake of cycloleucine (a nonmetabolizable analogue of leucine) by both muscle and liver was unaffected. (d) The percentage of a trace amount of injected [1-14C]leucine expired as 14CO2 in 1 h was significantly increased. (e) The capacity of muscle homogenate for α-decarboxylation of leucine was enhanced, whereas that of liver was unaffected. (f) The activity of leucine transaminase was unaffected, whereas that of α-ketoisocaproate dehydrogenase was increased in muscle.

Authors

Harbhajan S. Paul, Siamak A. Adibi

Thyrotropin-releasing Hormone Stimulation of Adrenocorticotropin Production by Mouse Pituitary Tumor Cells in Culture: POSSIBLE MODEL FOR ANOMALOUS RELEASE OF ADRENOCORTICOTROPIN BY THYROTROPIN-RELEASING HORMONE IN SOME PATIENTS WITH CUSHING'S DISEASE AND NELSON'S SYNDROME

• Text
• PDF

Abstract

ACTH-producing mouse pituitary tumor cells in culture (AtT-20/NYU-1 cells) were found to have binding sites for thyrotropin-releasing hormone (TRH). These putative receptors bound TRH with high affinity; the apparent equilibrium dissociation constant was 3.7 nM. The affinity of the receptors for a series of TRH analogues was similar to those previously reported for TRH-receptor interactions on thyrotropic and mammotropic cells in culture. Like some human pituitary tumors in situ, AtT-20/NYU-1 cells were found to produce the alpha subunit of the glycoprotein hormones (alpha). Alpha accumulation in the medium was constant (3.1 ng/mg cell protein per h) and was not affected by TRH. In contrast, TRH increased the amount of ACTH accumulated in the medium from AtT-20/NYU-1 cells to 190 and 420% of control at 1 and 24 h, respectively. TRH induced a dose-dependent increase in ACTH release during a 30-min incubation; half-maximal stimulation occurred at ∼0.1 nM. TRH had no effect on ACTH release in vitro from anterior pituitary cells derived from normal rats. Because TRH stimulates release of ACTH in some untreated patients with Cushing's disease and Nelson's syndrome as well as pathological states associated with pituitary tumors (but not in normal subjects), AtT-20/NYU-1 cells may serve as an important in vitro model for human pituitary ACTH-secreting adenomas. Moreover, these findings suggest that the primary abnormality in Cushing's disease and Nelson's syndrome, allowing TRH stimulation of ACTH release, may be intrinsic to neoplastic adrenocorticotrophs rather than in neuroregulation of ACTH release.

Authors

Marvin C. Gershengorn, Carlos O. Arevalo, Elizabeth Geras, Mario J. Rebecchi

Studies on the Pathogenesis of Pigment Gallstones in Hemolytic Anemia: DESCRIPTION AND CHARACTERISTICS OF A MOUSE MODEL

• Text
• PDF

Abstract

The pathogenesis of hemolysis-induced gallstones was studied in mice with a hereditary hemolytic disease called normoblastic anemia (genotype nb/nb) and in their normal controls (genotype +/+). Infrared spectroscopy demonstrated that spontaneously formed gallstones from nb/nb mice were nearly identical to stones from patients with chronic hemolysis as the result of sickle cell disease, and both mouse and human stones strikingly resembled synthetic calcium bilirubinate. 57% of 115 nb/nb mice, but none of 109 control mice, developed calcium bilirubinate pigment gallstones (P < 0.001). The incidence of luminal gallstones in nb/nb mice was both sex and age dependent. Female nb/nb mice formed stones twice as frequently as male nb/nb mice (P < 0.001). Before 6 mo of age neither sex developed stones, but thereafter the incidence of stones increased with age. Neither hematocrit, reticulocyte count, nor total plasma bilirubin values, were useful in distinguishing between nb/nb mice with or without gallstones. In gallbladder bile, nb/nb mice with gallstones had higher concentrations of hydrogen ion, total bilirubin, calcium, and bile acids than nb/nb mice without stones. Although total unconjugated bilirubin was similar in both nb/nb groups, the ionized fraction of unconjugated bilirubin was higher in bile from nb/nb mice without stones than those with stones. In nb/nb mice, neutral mucin plugs and pigment concentrations were observed histologically in the glandular crypts of the gallbladder in 33% of nb/nb mice without stones and in 80% of nb/nb mice with luminal stones. This suggested that luminal pigment stone disease in mice with hemolysis may be preceded by microscopic precipitation of calcium bilirubinate in the glandular crypts of the gallbladder. These precipitates may then migrate into the lumen and grow by accretion.

Authors

Bruce W. Trotman, Seldon E. Bernstein, Kevin E. Bove, Gary D. Wirt

Cytochemical Bioassay of Parathyroid Hormone: CHARACTERISTICS OF THE ASSAY AND ANALYSIS OF CIRCULATING HORMONAL FORMS

• Text
• PDF

Abstract

A cytochemical bioassay for parathyroid hormone (PTH) was used for the characterization of the biological activity of circulating forms of the hormone. PTH-stimulated glucose-6-phosphate dehydrogenase activity in distal convoluted tubule cells was quantitated by integrating microdensitometry and the response to native bovine (b)PTH(1-84) was found to be linear between graded doses of hormone from 5 fg/ml to 5 pg/ml. Synthetic bPTH(1-34) and human (h)PTH(1-34) elicited a parallel and equimolar response; however calcitonin, ACTH, glucagon, epinephrine, vasopressin, and insulin failed to significantly stimulate the enzyme in doses up to 100,000 times greater than the lowest concentration of bPTH used. The assay was capable of distinguishing hormonal activity in normal, hypoparathyroid, and hyperparathyroid human plasma. After gel chromatography, bioactivity in plasma of hyperparathyroid patients with skeletal disease but normal kidney function coeluted mainly with bPTH(1-84), whereas bioactivity in plasma of hyperparathyroid patients with skeletal disease but severe uremia coeluted in approximately equivalent amounts with bPTH(1-84) and hPTH(1-34). Despite the abundance of small molecular-weight bioactivity in the peripheral circulation in uremia, ∼85% of the bioactivity in the parathyroid venous effluent coeluted with bPTH(1-84). The results therefore demonstrate the sensitivity and specificity of the assay for PTH and its utility in measuring the hormone in human parathyroid disorders. The results furthermore demonstrate the importance of entities cochromatographing with bPTH(1-84) in comprising the circulating bioactive hormone in hyperparathyroidism, and support the concept of a biological role for smaller forms of PTH, at least in chronic renal failure.

Authors

David Goltzman, Brian Henderson, Nigel Loveridge

Variant von Willebrand's Disease: CHARACTERIZATION OF TWO SUBTYPES BY ANALYSIS OF MULTIMERIC COMPOSITION OF FACTOR VIII/VON WILLEBRAND FACTOR IN PLASMA AND PLATELETS

• Text
• PDF

Abstract

We have examined the multimeric composition of factor VIII/von Willebrand factor in plasma and platelet lysates by means of sodium dodecyl sulfate agarose electrophoresis followed by staining with 125I-labeled affinity-purified antibody. In normal plasma and platelet lysates, factor VIII/von Willebrand factor displayed 10 distinct multimers that ranged in apparent molecular weight from 0.86 to 9.9 × 106. The molecular weight difference between adjacent bands was 0.8−1.1 × 106. Larger material, not resolved into discrete bands, was also present with an average Mr of 14.5 × 106. Though the dimer (apparent Mr = 0.48 × 106) and the monomer (apparent Mr = 0.28 × 106) generated by reduction of disulfide bonds were readily identified in this system, they were not detected in normal plasma or platelets. No differences were observed between fresh plasma prepared without anticoagulant and fresh or frozen plasma anticoagulated with either citrate or heparin. “Variant” (type II) von Willebrand's disease could be divided into two subtypes. In subtype IIA, factor VIII/von Willebrand factor in plasma consisted predominantly of the five smaller multimers with traces of the sixth and seventh (Mr up to 4.5 × 106). In subtype IIB, all these multimers were easily detected and, in addition, bands of intermediate size (Mr = 8.5 × 106 and smaller) were present. In contrast, the multimeric composition of IIB platelet factor VIII/von Willebrand factor was identical to normal, whereas in subtype IIA the larger multimers were absent from platelets as well as from plasma. In subtype IIB, binding of factor VIII/von Willebrand factor to platelets occurred at lower concentrations of ristocetin than required for normal and multimers of smaller size than in normal bound. On the contrary, in subtype IIA, binding was minimal, as was true of normal factor VIII/von Willebrand factor of equivalent size. Thus, physical as well as functional differences in the two subtypes of variant von Willebrand's disease described suggest that different pathogenetic mechanisms underlie the factor VIII/von Willebrand factor abnormalities in these patients.

Authors

Zaverio M. Ruggeri, Theodore S. Zimmerman

Calcium and Inorganic Phosphate Transport in Rat Colon: DISSOCIATED RESPONSE TO 1,25-DIHYDROXYVITAMIN D₃

• Text
• PDF

Abstract

In the small intestine, 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] stimulates both calcium (Ca) and inorganic phosphate (Pi) absorption. This is mediated through an increase in mucosal-to-serosal flux (Jms) whereas the serosal-to-mucosal flux (Jsm) remains unchanged. We now report that in rat proximal colon, 1,25(OH)2D3 produces active Ca absorption without affecting Pi transport, and that this induced active Ca absorption is associated with alterations in kinetics of both Jms and Jsm so that both processes demonstrate saturable components. Vitamin D-deficient rats were given daily injections of solvent (−D) or 270 ng 1,25(OH)2D3 (+D) for 3 d. 45Ca and [32P]phosphate fluxes were measured employing the Ussing technique using a modified Krebs-Ringer-HCO3 buffer ([Ca] 1.25, [Pi] 1.18, [glucose] 11 mM). In −D rats there was no net flux (Jnet) of either Ca or Pi. In +D rats net active Ca absorption was observed (−D = 3.3 nmol/cm2 per h ±3.4 (SEM); +D = 27.3 ±3.8, n = 11, P < 0.001) whereas Pi transport was unchanged, i.e., still no Jnet. Pi Jms was not different from Pi Jsm measured at the following buffer [Pi]: 0.0118, 0.118, 1.18, and 2.36 mM. Ca saturation kinetics were estimated using buffer [Ca] from 0.0125 to 5.0 mM. Saturable processes were demonstrated for both Jms and Jsm. Jnet for Ca across colon from +D rats exhibited saturation at [Ca] > 3 mM, with an estimated Vmax of 44.0 nmol/cm2 per h and a Km of 0.9 mM. This colonic model may provide a useful system for studying 1,25(OH)2D3-induced molecular events related to Ca but not Pi transport. The apparent action of 1,25(OH)2D3 on Ca secretory process may furnish new insights into the mechanism of action of vitamin D.

Authors

D. B. N. Lee, M. W. Walling, U. Gafter, V. Silis, J. W. Coburn

Mechanism of Bilirubin Diglucuronide Formation in Intact Rats: BILIRUBIN DIGLUCURONIDE FORMATION IN VIVO

• Text
• PDF

Abstract

Although it is well established that bilirubin monoglucuronide is formed in the liver from bilirubin by a microsomal bilirubin uridine diphosphate (UDP)-glucuronosyltransferase, the subcellular site of conversion of monoglucuronide to diglucuronide and the molecular mechanism involved in diglucuronide synthesis have not been identified. Based on in vitro studies, it has been proposed that two fundamentally different enzyme systems may be involved in diglucuronide synthesis in rat liver: (a) a microsomal UDP-glucuronosyltransferase system requiring UDP-glucuronic acid as sugar donor or (b) a transglucuronidation mechanism that involves transfer of a glucuronosyl residue from one monoglucuronide molecule to another, catalyzed by a liver plasma membrane enzyme. To clarify the mechanism by which bilirubin monoglucuronide is converted in vivo to diglucuronide, three different experimental approaches were used. First, normal rats were injected with either equal amounts of bilirubin-IIIα [14C]monoglucuronide and unlabeled bilirubin-XIIIα monoglucuronide, or bilirubin-XIIIα [14C]monoglucuronide and unlabeled bilirubin-IIIα monoglucuronide. Analysis of radiolabeled diglucuronide excreted in bile showed that [14C]glucuronosyl residues were not transferred between monoglucuronide molecules. Second, in normal rats infused intravenously with dual-labeled [3H]bilirubin [14C]monoglucuronide, no transfer or exchange of the [14C]glucuronosyl group between injected and endogenously produced bilirubin monoglucuronide could be detected in the excreted bilirubin diglucuronide. Third, in homozygous Gunn rats, injected 14C-labeled or unlabeled bilirubin mono- or diglucuronides were excreted in bile unchanged (except that diglucuronide was hydrolyzed to a minor degree). This indicates that Gunn rats, which lack bilirubin UDP-glucuronosyltransferase activity, are unable to convert injected monoglucuronide to diglucuronide. Collectively, these findings establish that a transglucuronidation mechanism is not operational in vivo and support the concept that bilirubin diglucuronide is formed by a microsomal UDP-glucuronosyltransferase system.

Authors

Norbert Blanckaert, John Gollan, Rudi Schmid

Amiloride inhibits mammalian renal kallikrein and a kallikrein-like enzyme from toad bladder and skin.

• Text
• PDF

Abstract

Renal kallikrein is localized in luminal plasma membranes of the mammalian distal nephron and gains access to urine from this site. Its activity is regulated, in part, by aldosterone. These facts led us to study the effects of amiloride, a drug known to inhibit sodium reabsorption and potassium secretion at this site, on kallikrein activity. Amiloride inhibited the esterolytic activity of purified rat or human urinary kallikrein or of rat renal cortical cells upon a synthetic substrate (ID50 = 0.12-0.23 mM). Kinetic analyses showed that the enzyme inhibition was noncompetitive and reversible in nature. The kinin-generating activity of kallikrein acting upon kininogen substrates was also inhibited by amiloride, as measured by bioassay in the rat uterus of guinea pig ileum or by radioimmunoassay of liberated kinins (ID50 = 85 microM). No other diuretic drug tested inhibited kallikrein activity, except triamterene, which did so, weakly. In addition, kallikrein-like enzyme activity was discovered in the urinary bladder or skin of Bufo marinus toads and this activity was also inhibited by amiloride. The localization of the enzyme and its inhibition by this drug suggest that further study of relationships amongst the glandular kallikrein-kinen system and renal ion and water transport is warranted.

Authors

H S Margolius, J Chao

Extracellular Matrix and Control of Proliferation of Vascular Endothelial Cells

• Text
• PDF

Abstract

Bovine vascular endothelial cells plated at low cell density in the presence of high (10%) concentrations of serum and maintained on plastic tissue culture dishes proliferate slowly. If the cultures were exposed to fibroblast growth factors (FGF), the cells proliferated actively and, after a week, a monolayer composed of closely apposed and highly contact-inhibited mononucleated cells formed. In contrast to cultures maintained on plastic, cultures maintained on dishes coated with an extracellular matrix produced by corneal endothelial cells proliferated rapidly and no longer required FGF to reach confluence. Addition of FGF to such cultures did not decrease the mean doubling time, which was already at a minimum (18 h), nor did it result in a higher final cell density, which was already at a maximum (700-1,000 cells/mm2). Likewise, although human umbilical vein endothelial cells plated at low density on plastic did not proliferate, they proliferated rapidly when plated on dishes coated with an extracellular matrix. However, unlike bovine vascular endothelial cells, they still required FGF if the cultures were to become confluent.

Authors

Denis Gospodarowicz, Charles Ill

12-O-Tetradecanoyl-Phorbol-13-Acetate Release of Glycosyltransferases from Human Blood Cells

• Text
• PDF

Abstract

The mononuclear cells separated from human blood by Ficoll-Hypaque centrifugation contained and released sialyltransferase, galactosyltransferase, and fucosyltransferase. Granulocytes contained and released lesser amounts of glycosyltransferases, whereas platelets released more fucosyltransferase than sialyltransferase or galactosyltransferase. When mononuclear cells were incubated with 12-O-tetradecanoyl-phorbol-13-acetate (TPA), the release of these three glycosyltransferases increased two- to six-fold, and cell suspension glycosyltransferase activities decreased 10-50%. Mononuclear cells were fractionated into lymphocytes and monocytes using baby hamster kidney cells microexudate-coated flasks. TPA stimulated the release of glycosyltransferases from lymphocytes but not from monocytes. The release of glycosyltransferases by TPA-treated mononuclear cells was not further stimulated by reincubation with TPA and was not affected by puromycin, cAMP, or cGMP. Concanavalin A, a mitogenic stimulator of lymphocytes, also stimulated the release of glycosyltransferases from mononuclear cells, but to a lesser extent. TPA did not stimulate the release of 5′-nucleotidase or decrease its activity on the cell pellet. Triton X-100 (0.2%) stimulated the release of glycosyltransferases to the same extent as TPA, but also caused the release of 5′-nucleotidase. [3H]TPA bound specifically and reversibly to mononuclear cells. The possible relationship between glycosyltransferase release and TPA effect on the plasma membrane is discussed.

Authors

Chen-Kao Liu, Robert Schmied, Samuel Waxman

Constituents of Human Neutrophils that Mediate Enhanced Adherence to Surfaces: PURIFICATION AND IDENTIFICATION AS ACIDIC PROTEINS OF THE SPECIFIC GRANULES

• Text
• PDF

Abstract

The endogenous constituents of human neutrophils that enhance the adherence of the neutrophils to surfaces have been isolated from sonicates of purified neutrophils. The predominant adherence-enhancing activity in the neutrophil sonicates cofiltered on Sephadex G-75 with a major peak of chemotactic inhibitory activity and exhibited ∼30,000 mol wt. Sequential isoelectric focusing and electrophoresis in glycerol gradients of the 30,000-mol wt activities resolved two distinct acidic protein with isoelectric points of 3.6-3.8 and 3.3-3.4 that were designated the neutrophil adherence factor (NAF) I and II, respectively. Glutamic acid and aspartic acid together accounted for a total of 18 and 19% of the amino acids in purified preparations of NAF I and NAF II, respectively, whereas the basic amino acids lysine, arginine, and histidine represented <2 and 3% of the total residues. The preincubation of portions of 2 × 106 neutrophils with as little as 6 pmol of NAF I or 9 pmol of NAF II enhanced adherence to plastic petri dishes and inhibited chemotactic migration to a maximal extent, with comparable dose-response relationships for the two effects. Neither of the NAF was cytotoxic, exhibited substantial neutrophil chemotactic or chemokinetic activity, or influenced the phagocytosis of sheep erythrocytes sensitized with immunoglobulin (Ig)G. Analyses of subcellular fractions of neutrophils indicated that the NAF are contained predominantly in the specific granules. These distinctive acidic proteins of the specific granules of human neutrophils represent a new class of endogenous constituents that may regulate the involvement of neutrophils in inflammation.

Authors

Linda K. Bockenstedt, Edward J. Goetzl

Lysine Fluxes across the Jejunal Epithelium in Lysinuric Protein Intolerance

• Text
• PDF

Abstract

Lysinuric protein intolerance (LPI) is one of a group of genetic diseases in which intestinal absorption of the diamino acids lysine, arginine, and ornithine is impaired. In LPI, the clinical symptoms are more severe than in the kindred disorders. The mechanism of lysine absorption was, therefore, investigated in vitro on peroral jejunal biopsy specimens in seven patients with LPI and 27 controls. The lysine concentration ratio between cell compartment and medium was significantly higher in the LPI group (mean±SEM, 7.17±0.60) than in the controls (5.44±0.51). This was also true for the intracellular Na concentration (LPI, 73.6±10.8 mM; controls 42.3±3.7 mM). The rate of unidirectional influx of lysine across the luminal membrane was Na dependent and was the same in the two groups. In the absence of an electrochemical gradient, net transepithelial lysine secretion was observed in LPI. This was entirely the result of a 60% reduction of the unidirectional flux from mucosa to serosa. Calculation of unidirectional fluxes revealed the most striking difference at the basolateral membrane, where the flux from cells to serosa was reduced by 62% and the corresponding permeability coefficient reduced by 71%. A progressive reduction in short-circuit current appeared in the epithelia of all four patients with LPI tested after addition of 3 mM lysine. Thus, LPI appears to be the first disease in which a genetically determined transport defect has been demonstrated at the basolateral membrane.

Authors

J-F. Desjeux, J. Rajantie, O. Simell, A-M. Dumontier, J. Perheentupa

Effect of Portal Venous Blood Flow Diversion on Portal Pressure

• Text
• PDF

Abstract

To anticipate the hepatic vascular response to portacaval anastomosis, we studied portal pressure during diversion of portal blood through a temporary extracorporeal umbilical vein to saphenous vein shunt. The relationship of portal pressure to shunted flow was approximately linear. In five schistosomiasis patients (controls) portal diversion to 1,250 ml/min gave portal pressure-shunted flow curve slopes ranging from 0.13 to 0.57 cm water/100 ml per min (0.31±0.18, mean±SD). In 17 cirrhotic patients with portal hypertension a continuum of slopes was observed from within mean±2 SD of control (type A) to larger slopes (type B) indicating failure of portal pressure regulation. When portal flow was augmented by shunting from saphenous vein to portal vein, cirrhotic patients who had slopes less than mean±2 SD of controls during diversion (type A) exhibited a compliant system with small increases in portal pressure, whereas type B patients had significantly greater pressure increases. Selective investigations suggested that changes in portal pressure provoked compensatory changes in hepatic arterial blood flow that tended to maintain portal pressure at a set point. Type B patients demonstrated failure of this mechanism to varying degrees.

Authors

David S. Zimmon, Richard E. Kessler

Decreased Lysosomal Dipeptidyl Aminopeptidase I Activity in Cultured Human Skin Fibroblasts in Duchenne's Muscular Dystrophy

• Text
• PDF

Abstract

Several lysosomal enzymes were assayed in cultured human skin fibroblasts from patients with Duchenne's muscular dystrophy (DMD) and age- and sex-matched control patients (N). The activity of four glycosidases, cathepsin B1, and total autoproteolysis at pH 4.0 were unchanged between the groups, but dipeptidyl aminopeptidase I (DAP-I, or cathepsin C) in the DMD cells was found to be only 30% as active as in the control cells (P < 0.003). This difference is not the result of a redistribution or loss of enzyme during homogenization because the difference occurs in all homogenate fractions. DAP-I activity existing in N and DMD fibroblasts behaves identically with respect to activation by chloride ion, activation by the sulfhydryl reducing agent dithiothreitol, changes in hydrogen ion concentration (pH), changes in substrate concentration (i.e., apparent Km values), and changes in temperature (i.e., apparent activation energies). Mixtures of N and DMD cell sonicates display an additivity in DAP-I activity. These results support the conclusion that the catalytic function of the DAP-I molecule is equivalent between N and DMD fibroblasts, and that the decrease in tissue-specific DAP-I activity probably results from the fact that fewer enzyme molecules are present in the DMD cells. These results are also an indication that these nonmuscle cells are expressing some of the phenotypic aspects of the genetic defect in DMD. Cultured human skin fibroblasts may therefore be a useful cellular model in DMD research.

Authors

Benjamin B. Gelman, Linda Papa, Michael H. Davis, Eric Gruenstein

Transplacental stimulation of lung development in the fetal rabbit by 3,5-dimethyl-3'-isopropyl-L-thyronine.

• Text
• PDF

Abstract

The effect of thyroid hormone on maturation of fetal rabbit lung was studied with maternal treatment using 3,5-dimethyl-3'-isopropyl-L-thyronine (DIMIT), a synthetic analogue of triiodothyronine. To investigate the in vivo kinetics and distribution of DIMIT, we prepared [3H]DIMIT and injected both pregnant rats (18-21 d gestation) and rabbits (25 d gestation). In the rat, maximal concentrations of radioactivity in maternal plasma, fetal plasma, and amniotic fluid occurred within 10 min, 1-2 h, and 4-6 h, respectively, after intramuscular injection. After 7 h the concentration of radioactivity in fetal plasma was 163 and 71% of the maternal level in rats and rabbits, respectively, indicating that DIMIT readily crosses the placenta. We treated pregnant rabbits for 1-2 d with DIMIT in doses of 0.5-3 mg/kg per d and examined the fetuses at 26 and 27 d gestation. Treatment did not affect fetal growth or viability. In fetal liver, DIMIT increased the activity of NADPH cytochromeac reductase by 64% and decreased the glycogen content by 73% compared to controls. The rate of choline incorporation by lung minces increased in dose-dependent manner to a maximum of +104% at 3 mg/kg DIMIT; this does stimulated by 38% the activity of lung phosphatidic acid phosphatase (PAPase), a corticosteroid-responsive enzyme, but there was no increase in tissue PAPase activity at most lower doses of DIMIT that enhanced choline incorporation. Treated lungs had 38% less glycogen tha controls, but there was no effect on tissue levels of DNA, protein, or phospholipid. DIMIT treatment increased the amount of total phospholipid (+163%). saturated phosphatidylcholine (+330%), and PAPase activity (+134%) in lung lavage fluid. The DIMIT effects on both choline incorporation by lung minces and phospholipid content of lavage fluid were substantially greater than what had occurred with an optimal dose of betamethasone. DIMIT also increased corticosteroid binding capacity in fetal plasma and produced a dose-dependent increase (maximal threefold) in total and free corticoids of both maternal and fetal plasma. It is estimated that elevated endogenous corticoids probably account for less than one-third of the increases in phospholipid synthesis and secretion observed at the higher doses of DIMIT. These data indicate that administration of DIMIT to pregnant rabbits accelertes maturation of the surfactant system in fetal lung. The magnitude of the effects on phospholipid synthesis and secretion, along with the minimal effect of PAPase activity in fetal lung tissue, suggest that thyroid hormones affect different biochemical processes from those influenced by glucocorticoids.

Authors

P L Ballard, B J Benson, A Brehier, J P Carter, B M Kriz, E C Jorgensen

Cerebrotendinous xanthomatosis: a defect in mitochondrial 26-hydroxylation required for normal biosynthesis of cholic acid.

• Text
• PDF

Abstract

Oxidation of side chain of 5 beta-cholestane-3 alpha,7 alpha,12 alpha-triol was studied in a patient with cerebrotendinous xanthomatosis (CTX) and in control subjects, using various subcellular fractions of liver homogenate and a method based on isotope dilution-mass spectrometry. In the control, 5 beta-cholestane-3 alpha,7 alpha,12 alpha-triol was converted into 5 beta-cholestane-3 alpha,7 alpha,12 alpha,26-tetrol and 3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholestanoic acid by the mitochondrial fraction, and into 5 beta-cholestane-3 alpha,7 alpha,12 alpha,-25-tetrol by the microsomal fraction. In the CTX patient, liver mitochondria were completely devoid of 26-hydroxylase activity. The same mitochondrial fraction catalyzed 25-hydroxylation of vitamin D3. The microsomal fraction of liver of the subject with CTX contained more than 50-fold the normal amount of 5 beta-cholestane-3 alpha,7 alpha,12 alpha-triol. The basic metabolid defect in CTX appears to be a lack of the mitochondrial 26-hydroxylase. The excretion in the bile of 5 beta-cholestane-3 alpha,7 alpha,12 alpha,25-tetrol and 5 beta-cholestane-3 alpha,7 alpha,12 alpha,24 alpha,25-pentol observed in CTX patients may be secondary to the accumulation of the major substrate for the 26-hydroxylase, i. e., 5 beta-cholestane-3 alpha,7 alpha,12 alpha-triol, and exposure of this substrate to the normally less active microsomal 25-and 24-hydroxylases. It is concluded that the major pathway in the biosynthesis of cholic acid in human liver involves a mitochondrial C27-steroid 26-hydroxylation.

Authors

H Oftebro, I Björkhem, S Skrede, A Schreiner, J I Pederson

New Granulocyte Antigens Demonstrated by Microgranulocytotoxicity Assay

• Text
• PDF

Abstract

Although granulocyte transfusions and bone marrow transplantation are becoming common clinical modalities, our knowledge of surface nonerythroid, nonlymphoid, non-HLA hematopoetic antigens remains very incomplete. Accordingly, we have systematically screened sera from recipients of multiple granulocyte and whole blood transfusions, and immunoneutropenic patients for antibodies directed primarily at granulocytes.

Authors

J. S. Thompson, V. L. Overlin, J. M. Herbick, C. D. Severson, F. H. J. Claas, J. D. 'Amaro, C. P. Burns, R. G. Strauss, J. A. Koepke

DNA Content Analysis by Flow Cytometry and Cytogenetic Analysis in Mycosis Fungoides and Sézary Syndrome: DIAGNOSTIC AND PROGNOSTIC IMPLICATIONS

• Text
• PDF

Abstract

Flow cytometric (FCM) analysis of DNA content was performed on 82 lymph node and peripheral blood specimens from 46 patients with mycosis fungoides and the Sézary syndrome. Overall, 32 of the 46 patients (70%) had aneuploidy detected by FCM. Aneuploidy was present in 63% of the patients at the time of diagnosis before systemic therapy. In these patients, aneuploidy was frequently detected in blood and lymph node specimens scored as negative by cytology and histology, suggesting that unsuspected extracutaneous dissemination is present in many patients at the time of diagnosis.

Authors

Paul A. Bunn Jr., Jacqueline Whang-Peng, Desmond N. Carney, Mark L. Schlam, Turid Knutsen, Adi F. Gazdar

Purification and Biochemical Characterization of Nuclear Ribonucleoprotein Antigen using Purified Antibody from Serum of a Patient with Mixed Connective Tissue Disease

• Text
• PDF

Abstract

We have achieved a high degree of purification of nuclear ribonucleoprotein antigen from calf thymus nuclear extract through antibody affinity chromatography. Antibody to nuclear ribonucleoprotein was purified from the serum of a patient with mixed connective tissue disease and Sepharose 4B was covalently coupled with the purified human antibody. The sodium thiocyanate eluate from the affinity column contained active ribonucleoprotein antigen and the specific activity of the antigen in this eluate was 488 times higher than the original nuclear extract. The protein component of the eluate consisted of six polypeptides determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Two of them were shown to be antigenic by a hemagglutination inhibition test. The molecular weights of these two peptides were ∼13,000 and those of the other four were 13,000, 13,000, 30,000, and 65,000. The ribonucleic acid component of the eluate was shown by urea-polyacrylamide gel electrophoresis to contain five polynucleotides. Two of the five, estimated to contain 40 and 60 nucleosides, had antigenic activity. The other three polynucleotides which had more than 77 nucleosides had no antigenic activity. No modified nucleosides were found in these ribonucleic acid molecules. Even in this highly purified ribonucleoprotein antigen, Sm antigen was detected by immunodiffusion. This evidence indicated that there are some molecular associations between ribonucleoprotein and Sm antigens as has previously been suggested.

Authors

Makoto Takano, Paul F. Agris, Gordon C. Sharp

Subpopulations of circulating B cells and regulatory T cells involved in in vitro immunoglobulin E production in atopic patients with elevted serum immunoglobulin E.

• Text
• PDF

Abstract

The B lymphocyte subpopulations producing immunoglobulin (Ig)E and the regulatory T cells modulating this IgE production in normals, and in atopic patients with respiratory allergy, atopic dermatitis, and markedly elevated serum IgE levels (>5,000 ng/ml), were investigated. Peripheral blood lymphocytes (PBL) were separated into T and B cell fractions and the ability of B cells to produce IgE in the presence or absence of pokeweed mitogen (PWM) and/or T cells ws determined. The patients had a circulating population of cells which spontaneously produced up to 6 ng of IgE in vitro (per 4 X 10(5) non-E-rosetting cells) in the absence of T lymphocytes and PWM. PBL from normals did not possess such cells. This IgE synthesis occurred primarily (>75%) over the first 72 h of culture. There was a wide range in their activity between patients and from the same patient studied on repeated occasions (from <300 to 6,000 pg per culture). This spontaneous IgE production was inhibited by PWM (mean inhibition, 37%) or normal T lymphocytes (mean inhibition, 42%). The patients lacked T lymphocytes capable of inhibiting this spontaneous IgE synthesis in 7 of 13 experiments. Functionally distinct B cells were identified in the patients and normals that responded to PWM with IgE production in vitro and required T-helper cell activity. Patients had normal PWM-responsive B cell IgE biosynthetic activity and T-helper function for these B cells. Suppressor T cell activity for PWM-driven IgE synthesis was also evaluated. Both the normals' and the patients' T lymphocytes provided similar levels of T cell suppressor function for PWM-driven IgE production. Patients with elevated serum IgE possessed these inhibitory T cells at times when the T lymphocytes which suppressed spontaneous igE production were absent from their PBL.

Authors

A Saxon, C Morrow, R H Stevens

Virus Infection of Endothelial Cells Increases Granulocyte Adherence

• Text
• PDF

Abstract

Adherence of human granulocytes was measured on endothelial monolayers of human and bovine origin, grown in 35-mm Diam petri dishes and in cluster wells. Adherence to human endothelium in petri dishes using 1.0 ml of whole blood averaged 17.9±3.7%, and to bovine endothelium was 20.3±3.7%. Cluster wells required only 1/5 the endothelial cells needed for petri dishes, and 0.25 ml of whole blood yielded average adherence of 26.2±3.4 to human cells and 28.0±3.7 to bovine in the wells. The impact of infection of the endothelium by different viruses on subsequent granulocyte adherence was measured. Polio virus produced an acute lytic infection of human endothelial cells, with associated increased adherence to 185.4% of control 24 h after inoculation. Significantly increased adherence was noted at 6 h, before detectable cytopathic effect. Herpes simplex type I caused a similar rapidly lytic infection of bovine endothelium associated with increased adherence to 213.7% of control 6 h after inoculation. This augmented adherence could be demonstrated when granulocytes were suspended in physiologic saline solution, showing that antibody and complement need not be present. Trypsin treatment of infected monolayers did not prevent the augmentation, and supernate from infected monolayers increased the adherence of polymorphonuclear leukocytes to normal, uninfected monolayers. Chronic, slowly lytic infections, lasting 7 d or more, were induced with adenovirus in human endothelium and with measles virus in bovine cells. Adherence increased as virus was noted in the cell cultures on day 4, several days before cytotoxicity was seen. Thus, chronic viral infection of the endothelium appears possible, and results in increased granulocyte adherence. In naturally occurring disease, such an infection may act synergistically with adherent granulocytes to damage the endothelium, and may represent an in vitro model of vasculitis.

Authors

Rob Roy MacGregor, Harvey M. Friedman, Edward J. Macarak, Nicholas A. Kefalides

Myotonic muscular dystrophy: defective phospholipid metabolism in the erythrocyte plasma membrane.

• Text
• PDF

Abstract

Myotonic muscular dystrophy (MyD) is a systemic genetic disorder that is thought to result from a generalized cellular membrane defect although the exact nature of this defect is unknown. This study examines two calcium-dependent membrane processes that have been observed in erythrocytes from healthy individuals: calcium-stimulated phosphatidic acid accumulation and calcium-induced potassium leak. We find that erythrocytes from MyD patients, in contrast to controls, have markedly impaired phosphatidic acid accumulations while maintaining normal potassium leaks. The calcium uptakes and ATP contents of MyD erythrocytes are not different from controls. We conclude that phospholipid metabolism is altered in MyD erythrocytes. The specificity of this abnormality and its relationship to altered muscular function are not known.

Authors

J E Grey, H J Gitelman, A D Roses

Turnover of Lipoprotein (a) in Man

• Text
• PDF

Abstract

An elevated concentration of lipoprotein (a) [Lp(a)] in the serum has been considered a risk factor for coronary heart disease by various investigators. In the present study, the turnover of Lp(a) was investigated in nine individuals with serum Lp(a) levels ranging from 1 to 68 mg/100 ml. After intravenous injection of radioiodinated Lp(a), the radioactivity time-curve of the serum and the specific activitity time-curves of the isolated Lp(a) and Lp(a) apolipoproteins were measured for 14 d. More than 97% of the label was found in the protein moiety of Lp(a). During the entire study period, the serum radioactivity remained with Lp(a), only insignificant amounts of radioactivity were detectable in other lipoprotein fractions. The serum radioactivity time-curves and the specific activity time-curves of the isolated Lp(a) and Lp(a) apolipoproteins were identical.

Authors

Franz Krempler, Gerhard M. Kostner, Klaus Bolzano, Friedrich Sandhofer

Interaction of Cardiopulmonary and Somatic Reflexes in Humans

• Text
• PDF

Abstract

Activation of cardiopulmonary receptors with vagal afferents results predominantly in reflex inhibition of efferent sympathetic activity, whereas activation of somatic receptors reflexly increases sympathetic activity to the heart and circulation. Previous studies in experimental animals indicate that there is an important interaction between these excitatory and inhibitory reflexes in the control of the renal circulation.

Authors

John L. Walker, Francois M. Abboud, Allyn L. Mark, Marc D. Thames

In vivo oxygen transport in the normal rabbit femoral arterial wall.

• Text
• PDF

Abstract

In vivo measurements of tissue oxygen tension were made at 10-micrometer intervals through functioning in situ rabbit femoral arterial walls, using inhalation anesthesia and recessed microcathodes with approximately 4-micrometer external diameters. External environment was controlled with a superfusion well at 30 torr PO2, 35 torr PCO2. Blood pressure, gas tension levels, and blood pH were held within the normal range. Radial PO2 measurements closely fit a mathematical model for unidimensional diffusion into a thick-walled artery with uniform oxygen consumption, and the distances traversed fit measured dimensions of quick-frozen in vivo sections. Using standard values of diffusion and solubility coefficients, mean calculated medial oxygen consumption was 99 nl0/ml-s. Mural oxygen consumption appeared to be related linearly to mean tangential wall stress. Differences in experimental design and technique were compared with previous in vivo and in vitro measurements of wall oxygenation, and largely account for the varying results obtained. Control of environment external to the artery, and maintenance of normally flowing blood in the lumen in vivo appeared critical to an understanding of mural oxygenation in life. If the conditions of this experiment prevailed in arteries with thicker avascular layers, PO2 could have been 20 torr at approximately 156 micrometer and 10 torr at 168 micrometer from blood (average values).

Authors

D W Crawford, L H Back, M A Cole

Hapten-specific T-Cell Unresponsiveness Induced by Benzylpenicilloyl Autologous Gamma Globulin Conjugates in Human Lymphocytes In Vitro

• Text
• PDF

Abstract

The aim of these studies was to determine whether unresponsiveness to the main determinant of penicillin, benzylpenicilloyl, can be induced in human peripheral lymphocytes in vitro by conjugates of benzylpenicilloyl (BPO) autologous gamma globulin (HGG). Initially it was shown that conjugates of BPO-keyhole limpet hemocyanin (KLH) elicited lymphocyte proliferation in the peripheral blood lymphocytes of six out of nine adult individuals in vitro. In contrast, conjugates of dinitrophenylated KLH and of BPO-HGG and the carriers HGG and KLH alone failed to do so. Similarly, release of the non-specific helper factor, lymphocyte mitogenic factor (LMF) occurred only after BPO-KLH stimulation. LMF activity was measured by B-cell proliferation and incorporation of radioactive amino acids into secreted immunoglobulin.

Authors

Raif S. Geha, Lazar Fruchter, Yves Borel

Protection of Human Neutrophils by Endogenous Catalase: STUDIES WITH CELLS FROM CATALASE-DEFICIENT INDIVIDUALS

• Text
• PDF

Abstract

To investigate the importance of catalase as a protecting enzyme against oxidative damage in phagocytic leukocytes, we have tested the functional capacity of neutrophils from two individuals homozygous for Swiss-type acatalasemia and from two individuals heterozygous for this deficiency. In the former cells, 25-30% of residual activity of catalase was present. In the latter cells, the values were close to normal.

Authors

Dirk Roos, Ron S. Weening, Sonja R. Wyss, Hugo E. Aebi

Parathyroid hormone acutely increases polyphosphoinositides of the rabbit kidney cortex by a cycloheximide-sensitive process.

• Text
• PDF

Abstract

Parathyroid hormone(PTH) rapidly increases the concentrations of diphosphoinositide and triphosphoinositide in rabbit kidney cortex. Cycloheximide pretreatment abolishes these effects of PTH. These findings are similar to those reported for adrenocorticotropin and cyclic AMP action in the adrenal cortex, and suggest a common mechanism. Cycloheximide-sensitive effects of PTH, e.g., phosphaturia, may require polyphosphoinositides and/or other phospholipids.

Authors

R V Farese, P Bidot-López, A Sabir, J S Smith, B Schinbeckler, R Larson

Autologous rosette-forming T cells as the responding cells in human autologous mixed-lymphocyte reaction.

• Text
• PDF

Abstract

Autologous rosette-forming cells (Tar cells) have surface and functional characteristics of post-thymic precursors and among these characteristics there are some that have been identified in the responsive cell of the autologous mixed-lymphocyte reaction (AMLR). We therefore did AMLR with circulating mononuclear cells from normal subjects using as responding cells either total T cells, T cells depleted of Tar cells, or purified Tar cells. The response of Tar cells in AMLR was significantly greater than that of total T cells and these responded significantly more than Tar-depleted T cells. Conversely, Tar cells responded less than total T cells or T cells depleted of Tar cells in allogeneic mixed-lymphocyte reactions. Increasing numbers of Tar cells gave significantly greater AMLR responses both alone and when added to diminishing proportions of Tar-depleted T cells to keep the number of T cells constant in the system. Tar cells are the responding cells in AMLR but not in allogeneic mixed-lymphocyte reactions.

Authors

R Palacios, L Llorente, D Alarcón-Segovia, A Ruíz-Arguelles, E Díaz-Jouanen

Inhibitory action of a met-enkephalin on ACTH release in man.

• Text
• PDF

Abstract

In order to study the mechanism of action of a Met-enkephalin (FK 33824) on the pituitary-adrenal axis eight normal male volunteers were subjected to an ACTH stimulation test. Lysine-vasopressing (LVP), 5 IU, was injected intramuscularly after pretreatment with 0.5 mg FK 33824 i.m. or a placebo. In six of the subjects the opiate was again administered preceding a single injection of 0.25 mg ACTH beta 1-24 i.m. Blood was collected at regular intervals and ACTH and cortisol concentrations analyzed in all samples. LVP induced significant plasma ACTH (P < 0.05) and cortisol (P < 0.001) increases. Pretreatment with FK 33824 completely antagonized the effect of LVP. Furthermore, the cortisol elevation after exogenous ACTH was not modified by previous administration of FK 33824. It is concluded that the Met-enkephalin derivative FK 33824 directly suppresses ACTH release from the pituitary without influencing adrenal synthesis of cortisol.

Authors

E del Pozo, J Martin-Perez, A Stadelmann, J Girard, J Brownell