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[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

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

[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

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

[The development of the adrenal cortex in pig (Sus scrofa domestica). (A contribution on the development of the fetal interior zone of the adrenal cortex in pig) (author's transl)].

The adrenals of pigs have been investigated histologically and histochemically during different periods of time pre- and postnatally. The pig (Sus scrofa domestica) shows a proper fetal cortex in the prenatal development of the adrenal gland. This involves slowly during the last quarter of pregnancy. A weight reduction is not noted thereby. Just before delivery the adrenal cortex of the pig has been subdivided into a zona arcuata and zona fasciculata. The zona reticularis develops only postnatally.

Adrenal Cortex

The mitochondrial malic enzymes. I. Submitochondrial localization and purification and properties of the NAD(P)+-dependent enzyme from adrenal cortex.

Rat and calf adrenal cortex homogenates were found to contain three different malic enzymes. Two were strictly NADP+-dependent and were localized, one each, in the cytosol and the mitochondrial fractions, respectively. These two enzymes appear to be identical to those described by Simpson and Estabrook (Simpson, E. R., and Estabrook, R. W. (1969) Arch. Biochem. Biophys. 129, 384-395). The third was NAD(P)+-linked and was present in the mitochondrial fraction only. All three malic enzymes separated as distinct bands during electrophoresis on 5 percent polyacrylamide slab gels at pH 9.0. Marker enzymes and the mitochondrial malic enzymes migrated together in intact mitochondria during sucrose density gradient centrifugations despite changes in the equilibrium position of the mitochondria promoted by energy-dependent calcium phosphate accumulation. In adrenal cortex mitochondria subfractionated by the method of Sottocasa et al. (SOTTOCASA, G.L., KUYLENSTIERNA, B., ERNSTER, L., and BERGSTAND, A. (1967) J. Cell Biol. 32, 415-438), both malic enzymes were associated with the inner membrane-matrix space. Sonication solubilized the two malic enzymes along with the matrix space marker enzymes. The NAD(P)+-dependent malic enzyme was purified 100-fold from calf adrenal cortex mitochondria. The final preparation was free of malic dehydrogenase, fumarase, the strictly NADP+-linked malic enzyme and adenylate kinase. Either Mn24 orMg2+ was required for activity and 1 mol of pyruvate was formed for each mole of NAD+ and NADP+ reduced. The pH optima with NAD+ and NADP+ were 6.5 tp 7.0 and 6.0 to 6.5, respectively. Michaelis-Menten kinetics were observed on the alkaline side. Fumarate, succinate, and isocitrate were positive and ATP and ADP were negative modulators of the regulatory enzyme. The modulators did not influence the stoichiometry and they were not metabolized during the reaction. Under Vmax conditions the ratios for the rate of NAD+:NADP+ reduction were 1.76 and 1.15 at pH 7.4 and 6.0, respectively. The apparent Michaelis constants also differed depending on the pH and the coenzyme. At pH 7.4 (in the presence of 5 mM fumarate) and at pH 6.0 (no fumarate) the Km values for (-)-malate, NAD+, and Mn2+ were 1.7, 0.16, and 0.15 mM, and 0.31, 0.06, and 0.09 mM, respectively. At pH 7.4 (5MM fumarate) and pH 6.0 (no fumarate), the Km values for (-)-malate, NADP+, and Mn2+ were 6.5, 0.62, and 0.59 mM, and 0.68. 0.12, and 0.31 mM, respectively. The apparent Ki values for ATP with NAD+ and NADP+ as coenzyme were 0.42 and 0.27 mM, respectively.

Adrenal Cortex

Specific lamellar structures of agranular endoplasmic reticulum in the senile mouse adrenal cortex.

In the adrenal cortex of old mice, lamellar bodies of various shapes frequently appear in the zona reticularis cells. These structures consist of electron dense plates, about 20 nm in width, closely piled at regular distances of about 25 nm. Each plate displays two sheets of unit membranes enclosing a dense material and with several clear spaces between them. These paired membranes usually terminate in round saccules at both ends, but occasionally continue to the tubules of surrounding agranular endoplasmic reticulum. Saccules in several rows, in combination with a part of the lamellae, may be arranged cirularly in the cytoplasm. From these observations, it is proposed that the lamellar body is composed of a cup-shaped mass of flattened cisternae of the agranular endoplasmic reticulum and each of the cisternae is attenuated and closed except at their marginal sacculations. The concentric or horseshoe-shaped profile of the lamellar body observed in this study is accounted for by different section planes through the cup-shaped structure.

Adrenal Cortex

Oestrogen induced hypertriglyceridaemia: role of the adrenal cortex.

The role of the adrenal cortex in the pathogenesis of hypertriglyceridaemia associated with the intake of oral contraceptive agents containing oestrogen has been investigated in rats. Bilateral adrenalectomy reduced the activity of hepatic enzymes regulating lipogenesis (acetyl CoA carboxylase, fatty acid synthetase) and decreased plasma triglyceride concentrations. On the other hand, the administration of high dosage corticosterone induced the activity of hepatic enzymes with consequent elevation in serum triglyceride levels. In animals with intact adrenals the administration of oestradiol: (a) raised plasma triglyceride levels, (b) enhanced the activity of hepatic enzymes, and (c) increased the adrenal cortex:body weight ratio. The effects (a) and (b) were not observed when both adrenals were removed prior to oestrogen therapy. High dosage corticosterone replacement was found to be essential for the oestradiol to produce its effects on hepatic enzymes and plasma triglyceride levels. The results suggest a regulatory role for the adrenal cortex in the homeostasis of plasma triglyceride concentration and that the hypertriglyceridaemia induced by the oestrogen containing preparations might be secondary to alterations in adrenocortical function.

Acetyl-CoA Carboxylase

[Glucocorticoid and androgenic functions of the adrenal cortex and the state of the sympathetic-adrenal system in thyrotoxicosis with primary organic disease of the central nervous system].

In patients with thyrotoxicosis combined with primary affection of the central nervous system (CNS)-encephalitis, diencephalitis-there was revealed, like in common thyrotoxicosis, an increased production by the adrenal cortex of hydrocortisone, its intensified transformation and inactivation, reduced production of 17-ketosteroid chiefly due to the fall of dehydroepiandrosterone content, an increased urinary excretion of adrenaline and a decreased-of noradrenaline. The use of functional test with the ACTH administration demonstrated organic affection of the CNS to sharply aggravate the weakening and even the exhaustion of the functional reserves of the glomerular and the reticular zones of the adrenal cortex developing during thyrotoxicosis, and also the reserve possibilities of the sympathico-adrenal system. Direction of the changes in the 24-hour excretion of 17-OCS and corticosteriods was the same in the majority of cases. The latter can point to the presence of functional association between the hypothalamus-hypophysis-adrenal cortex systems and adrenergic regulation in these patients.

17-Hydroxycorticosteroids

Functioning and nonfunctioning cysts of the adrenal cortex and medulla.

Adrenal cysts are rare clinical and pathologic entities. The vast majority are minute in size, unilateral, and found in females at autopsy. They come to the physician's attention when they produce symptoms or are identified on x-ray studies. When symptomatic, they generally produce lumbar discomfort and gastrointestinal symptoms, and occasionally are palpable by abdominal examination. There is no significant agreement as to cause and pathogenesis of the disease. The most common histologic types are lymphangiomatous endothelial cysts, secondary to lymphangiectasis, and the fibrous wall or hemorrhagic pseudocysts. In many patients preoperative diagnosis is now possible with high quality nephrotomography, ultrasonography, and adrenal arteriography. The presence of a suprarenal mass with peripheral or laminar calcification strongly suggests the presence of an adrenal pseudocyst. Selective biochemical studies are mandatory to rule out the presence of an occult cystic pheochromocytoma, and adrenal cortical tumor. Adrenal cysts must be differentiated from all space-occupying lesions of the upper abdomen. Surgical exploration is recommended in almost all patients for accurate diagnosis and to rule out malignant disease or occult pheochromocytoma. The choice of the surgical approach should be planned to provide for safe and adequate exposure, depending on the size and location of the lesion. Careful dissection with preservation of the adjacent kidney, liver, and pancreas should be performed.

Adrenal Cortex