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Nuclear bodies in the normal and hyperfunctional human adrenal cortex.

Adrenal pieces obtained from six female patients, three without increased adrenocortical function and three with Cushing's disease, showed, in all adrenal cortex zones, cells containing simple and complex nuclear bodies. The simple nuclear bodies were spherical or ovoid and had a filamentous structure surrounded by a clear halo. Complex nuclear bodies were more numerous and heterogeneous in patients with adrenal pathology, and they were spherical with a proteinaceous filamentous capsule surrounding a core; the core was granular, filamentous or a mixture of granular and filamentous material, sometimes with a reticular or concentric arrangement. Some bodies showed vacuolar or multilocular aspect, and others had a close relationship with the nucleolus or appeared near the interchromatin granules. The meaning of adrenal nuclear bodies is discussed as well as their relationship with ACTH stimulation.

Adolescent

Endocrine emergencies. Disorders of the adrenal cortex.

Adrenal crisis represents the major endocrine emergency with a fatal outcome if not properly recognized and correctly treated. It can present as the first manifestation of acute or chronic primary adrenal failure. Secondary adrenal insufficiency with usually less dramatic manifestations can cause mental disturbances, electrolyte or metabolic disorders severe enough to warrant prompt correction and therapeutic intervention. This is particularly true in cases of abrupt cessation of steroid treatment, pituitary apoplexy and Sheehan's syndrome. Hyperfunction of the adrenal cortex, be it hypercorticism or hypermineralocorticism, can also require rapid treatment of electrolyte disorders and hypertension. The symptoms and differential diagnosis of these entities are described as well as their proper treatment. In order to confirm later the diagnosis, the need to secure biological samples for hormone determination before starting treatment is emphasized.

Acute Disease

Transplantation of parathyroid, adrenal cortex and adrenal medulla using procedures which successfully prolonged islet allograft survival.

Life-long hormone replacement therapy is today the only form of therapy for several endocrine insufficiencies. Transplantation of endocrine tissues could represent a more physiological approach to the treatment of different syndromes such as adrenal insufficiency, diabetes or hypoparathyroidism. Successful experimental islet transplantation across major histocompatibility barriers using cultural pretreatment of the insulin producing tissue in conjunction with temporary immunosuppression of the recipients, encouraged us to attempt similar approaches with parathyroids, adrenal cortex and adrenal medulla. Either bilateral parathyroidectomy or bilateral adrenalectomy were performed on the recipient rats before transplantation, followed by the implantation of parathyroid, adrenal cortical or adrenal medullary tissue beneath the renal capsule. The tissue was transplanted either immediately after its dissection or after pretreatment with 1 week culture. Some of the recipients received a short term cyclosporine treatment. The results indicated that parathyroid and adrenal medulla transplants were often infiltrated by mononuclear cells 30 days after transplantation. In contrast, adrenal cortical tissue was well preserved 4 weeks after transplantation, without showing any sign of rejection when a combination of pre-transplant culture of the adrenal tissue and temporary cyclosporine treatment of the recipients was used.

Adrenal Cortex

Effect of constant light and darkness on the mitotic activity of adrenal cortex undergoing adrenal regeneration or compensatory growth in male Wistar rats.

The effect of constant light and darkness on the mitotic activity of the adrenal cortex in the course of adrenal regeneration and compensatory growth in male Wistar rats was investigated. Three separate experiments were performed in different seasons. It was found, that on the tenth day of both regeneration and compensatory growth, constant darkness decreased the mitotic activity of the adrenal cortex, and in contrast, constant light had no significant effect when compared with the controls.

Adrenal Cortex

[Interrelationship between the quality of dietary protein and the function of the adrenal cortex. 3. Reactivity of the adrenal cortex and ACTH content of the pituitary gland of rats fed with proteins of various quality].

Trials were performed investigating the reactivity of the adrenals and the ACTH content of the pituitary gland in rats fed proteins of varying quality. A positive correlation was found to exist between the relative weight of the adrenals and the rate of body weight increases as well as between the weight of the pituitary gland and the rate of body weight gains. In either case this process was found to be primarily dependent on the quality of the dietary protein. It was shown that the quality of the dietary protein did not affect the in vitro reactivity of the adrenal glands. The ACTH content of the pituitary gland( determined on the basis of 100 mg of pituitary tissue) did not change considerably.

Adrenal Cortex

[Process of bile formation and bile composition in albino rats in acute adrenal cortex insufficiency and adrenal gland autologous transplantation].

Experiments were conducted on 84 male albino rats. A study was made of the state of the bile forming process 7, 14 and 30 days after bilateral adrenalectomy and adrenal gland autotransplantation. Investigations carried out showed inhibition of the bile formation with the corresponding changes in the chemical composition of bile to occur both in adrenalectomized animals and the rats with adrenal autotransplantation; in the second case, however, these changes were less pronounced. A gradual suppression of the intensity of bile production, cholate formation, bilirubin- and cholesterol secretion occurred in the adrenalectomized rats, whereas in the animals a significant shift in the direction of restoration took place as soon as in 7 days after autotransplantation. The first to become restored is the cholesterin secretory function in the liver, then the intensity of the cholate formation, particularly the glycocholic and desoxycholic acids, and bilirubin secretion. The intensity of bile secretion, as well as the sum total content of cholates in the bile, chiefly on account of taurocholic acid, failed to reach the initial level even by the 30th day after the adrenal gland autotransplantation.

Acute Disease

Zonation of the adrenal cortex. I. Isolation of mitochondria from the zona glomerulosa of the bovine adrenal cortex.

Isolation procedures for mitochondria from the zona glomerulosa of the bovine adrenal cortex are described in comparison to those isolated from the zona fasciculo-reticularis. The cristal membranes of mitochondria in the zona glomerulosa in situ are tubular or tubulo-vesicular, whereas those of mitochondria in the zona fasciculo-reticularis in situ are vesicular. When mitochondria are isolated from the former zone, they invariably showed a condensed configuration regardless of isolation media, whereas those isolated from the later zone in a ST medium showed an orthodox configuration. When Ca2+ was added to mitochondria isolated form either zone in an STE medium in the condensed configuration, transition from the condensed to the orthodox configuration took place; the cristel membrane of mitochondria from the zona glomerulosa became tubular or tubulo-vesicular and those of mitochondria from the zona fasciculo-reticularis became vesicular. Purity of mitochondria thus obtained from the zona glomerulosa was examined by electron microscope and marker enzymes. Coupling efficiency of mitochondria was found to be remarkably affected by temperature during the isolation procedures and a choice of substrates.

Adrenal Cortex

Pregnenolone-binding proteins in the rat adrenal cortex.

Rat adrenal cortex contains a protein(s) that binds pregnenolone with high affinity. This binding, demonstrated by gel filtration and by a dextran-coated charcoal method, was associated with the cytosol and with fractions solubilized by sonication from the mitochondria and microsomes. The binding of pregnenolone was saturable and was inhibited by mercurials and proteolytic enzymes. Pregnenolone-binding was not influenced by the presence of progesterone, deoxycorticosterone, corticosterone or aldosterone, but was inhibited by steroids with a 3beta-hydroxy-5-ene structure similar to pregnenolone, and by hydroxymethylene steroid and cyanoketone. We suggest that this protein is involved in the intracellular transport and retention of pregnenolone within adrenal cortical cells.

Adrenal Cortex

Steroidogenesis in the zona glomerulosa of the adrenal cortex. II. Distribution of cytochrome P-450 in the zona glomerulosa of the bovine adrenal cortex.

Microsomes were obtained from the zona glomerulosa of the bovine adrenal cortex. Contamination of microsomes with other cellular organelles was examined using various marker enzymes and the electron microscope. Distribution of cytochrome P-450 in mitochondria and in microsomes was determined to be 0.73 and 0.32 nmol/mg protein, respectively. The CO difference spectrum was affected not only by the concentration of added deoxycholate but also by the incubation time after addition. Approximately 40-50% of cytochrome P-450 in the samples was converted to cytochrome P-420 within 20-30 sec of incubation with deoxycholate. The content of RNA, phospholipids, and cytochrome b5 in microsomes obtained from the zona glomerulosa is also evaluated in comparison to that in microsomes obtained from the zona fasciculoreticularis.

Adrenal Cortex

Zonation of the adrenal cortex. III. Distribution of cytochrome P-450 in the zona glomerulosa of the bovine adrenal cortex.

A microsomal fraction was obtained from the zona glomerulosa of the bovine adrenal cortex. Glucose-6-phosphate activity of the fraction was found to be much lower than that of the liver. Contents of RNA and phospholipids, besides electron microscopic findings, of the fraction also indicate that it is rich in smooth-surfaced endoplasmic reticulum. Distribution of cytochrome P-450 in the zona glomerulosa was studied using various fractions including the microsomal fraction described above. The amount of cytochrome P-450 in mitochondria and that in microsomes were determined to be 0.73 and 0.32 nmoles/mg protein, respectively. The CO-difference spectrum was affected not only by the concentration of added deoxycholate but also by the incubation time after addition. Approximately 40-50% of cytochrome P-450 in the samples were converted to cytochrome P-420 within 20-30 seconds of incubation with deoxycholate.

Adrenal Cortex

[Relationship between food protein quality and adrenal cortex function. 6. Interrelationship between aminotransferase activity (GPT1, GOT2) and the status of adrenal cortex function].

Growing male Albino rats were adrenalectomized and then treated with corticosterone in an attempt to determine the activity of amino transferases (GPT and GOT) in the liver and in the mucosa of the small intestine over a period during which the animals received dietary proteins of varying quality. However, statistically significant differences in the enzyme activity were established for only those groups in which the animals exhibited differences in the functional state of the adrenal cortex. Differences due to changes in the quality of the dietary protein were not observed. From this was concluded that the activity of the amino transferases in the tissues is largely dependent on the action of corticosteroids.

Adrenal Cortex

Adrenal cortex adenylate cyclase. Specific binding sites for 5'-guanylyl-imidodiphosphate in partially purified plasma membranes from bovine adrenal cortex.

Specific binding sites for 5'-guanylyl-imidodiphosphate [Gpp(NG)p] have been identified in a partially purified plasma membrane fraction from bovine adrenal cortex. The apparent affinity of Gpp(nh)p at 30 degrees C was 12muM-1 and the concentration of binding sites was 100 pmoles per mg of protein. Binding of Gpp(NH)p is inhibited by Mn2+ greater than Mg2+ greater than Ca2+ and enhanced by low concentrations of the chelators ethylenediamino-tetraacetic acid (EDTA) and ethylene glycolbis-(beta-aminoethylether))-N,N'-tetraacetic acid (EGTA). High concentrations of EDTA are inhibitory and at 2.5 mM EDTA binding of Gpp(NH)p is only 10% of that observed in the absence of the chelator. The bound labeled GTP analogue exchanged only slowly with the unlabeled nucleotide after a steady state has been reached. EDTA also releases the bound labeled Gpp(NH)p from its binding sites. The slow dissociation of Gpp(NH)p can explain the persistent activation of adenylate cyclase observed after pretreatment of bovine adrenal cortex plasma membranes with Gpp(NH)p and subsequent washing. It is suggested that at least parts of these binding sites are identical to the sites identified earlier as regulatory sites for angiotensin high-affinity receptors (Glossmann et. al., 1974a) and for ACTH-stimulated cyclase Glossmann and Gips, 1974).

Adenylyl Cyclases

[Relationship between food protein quality and adrenal cortex function. 7. Relationship between activity of selected liver and small intestine hydrolases and adrenal cortex function].

Male Wistar rats received 10% true digestible crude protein of varying quality (maize gluten (MK) = low protein quality; maize gluten + lysine, tryptophan, threonine, isoleucine (MK + AS) = high protein quality) for a period of three weeks. The trial was carried out to investigate the activity of the two enzymes leucine-arylamidase (LNA) and aminotripeptidase (TP) in the liver and the mucosa of the small intestine and that of alpha and gamma amylases and of total amylases in the mucosa of the small intestine in animals exhibiting differences in the functional state of their adrenal cortices (adrenalectomized animals (AE), normal rats (N) and animals receiving additional treatment with cortisol (KS). It was found that mucosal LNA and gamma amylase did not respond to functional changes in the adrenal cortex. Mucosal LNA had a higher activity if the dietary protein was of low quality and a lower activity if the protein quality was high. The level of liver LNA activity was significantly lower in N and KS animals than in AE animals. The highest activity of mucosal and liver TP was found in KS animals; it was lowest in N animals. Statistically significant differences brought about by changes in the quality of dietary proteins were only found to occur in the KS group. The activity of alpha amylase and total amylases was found to be lowest in N animals; it was significantly higher in animals of the AE and KS groups. Animals of the N and KS groups receiving MK + AS exhibited a significantly higher level in their alpha and total amylase activities than animals in the same group receiving MK. Here, the "permissive action" of glucocorticosteroids on metabolically active hydrolases is clearly evident. The pattern of mucosal LNA activity in the small intestine and that of gamma amylase may be regarded as further characteristic evidence for the digestive action of these enzymes. Liver LNA and liver TP as well as mucosal TP and alpha amylase acted like metabolic enzymes.

Adrenal Cortex

Expression of an abundant alpha-class glutathione S-transferase in bovine and human adrenal cortex tissues.

Bovine adrenal cortex tissue expresses high levels of glutathione S-transferase (GST) from each of the alpha, mu and pi gene families. We describe the purification and characterization of an abundant alpha-class GST from this tissue that has not been identified previously because of its failure to bind to S-hexylglutathione-Sepharose 6B (S-hexG-Ag). This enzyme has been affinity purified on glutathione-Sepharose 6B (GSH-Ag) and was obtained in a highly purified form by employing S-hexG-Ag to remove the bulk of GST before chromatography on GSH-Ag. The purified GST eluted from GSH-Ag was found to exhibit marked peroxidase and delta 5-ketosteroid isomerase activities (19.2 and 1.67 U/mg respectively). The bovine enzyme also showed high GST activity towards 4-hydroxynonenal (5.09 U/mg). Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed that the bovine GST contains two distinct polypeptides, one with an Mr of 25,900 and the other with an Mr of 26,500. An abundant alpha-class GST was also purified from human adrenal cortex that possessed properties which were similar to the bovine alpha-class GST described above; however, unlike the bovine enzyme, the corresponding human alpha-class GST bound to S-hexG-Ag. As with the bovine enzyme, the purified human GST displayed marked peroxidase and isomerase activities (27 and 4.02 U/mg respectively). Further analysis on SDS-PAGE (Mr 25,800) and reverse-phase high-performance liquid chromatography established that this abundant alpha-class GST in human adrenal cortex is equivalent to the human liver GST B1B1 enzyme. As both human and bovine adrenal cortex contain high levels of alpha-class GST with similar catalytic properties, we discuss the possible functions of these enzymes in this tissue.

Adrenal Cortex

Zonation of the adrenal cortex. II. Effect of BSA on coupling efficiency of mitochondria isolated from the zona glomerulosa of the bovine adrenal cortex.

Effect of bovine serum albumin on coupling efficiency of mitochondria isolated from the zona glomerulosa of the bovine adrenal cortex in various media was examined polarographically and electron microscopically. Albumin restored the coupling efficiency of mitochondria isolated from the zona glomerulosa regardless of isolation media when succinate or malate was oxidizable substrate. Respiratory controls greater than 5 were obtained. Albumin, however, had no effect when glutamate, beta-hydroxybutylate and pyruvate were the oxidizable substrates. The conditions have been found under which mitochondria of the zona glomerulosa stay in the orthodox configuration and yet coupled.

Adrenal Cortex