[THOUGHTS ON THE MECHANISM OF ACTION OF THE THERAPEUTIC EFFECT FOLLOWING INTRA-ARTICULAR APPLICATION OF ADRENAL CORTEX DERIVATIVES].
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The authors analyzed the results of examination of 69 patients operated on for Itsenko-Cushing's syndrome (ICS); in 32 cases tumours of the adrenal cortex were revealed during the operation. Two histological forms of adenoma of the adrenal cortex were distinguished, i.e. adenomas of mono- and polymorphous structure. Increased background urinary excretion of 17-oxycorticosteroids (17-OCS) in patients with ICS suffering from tumours of polymorphous structure persisted at the former level or increased in the tests with 8 mg of dexamethazone (DMZ). The character of tests with 8 mg of DMZ in patients with ICS having adenomas of monomorphous structure was analogous to such in patients with ICS, but without any adrenal gland tumours--urinary 17-OCS excretion was either decreased or suppressed by 50 or more per cent of the initial level. A suggestion is put forward on functional dependence of the adrenal cortex of monomorphous structure on the hypothalamo-hypophysial control.
1. Specific [3H]estradiol binding activity with characteristics of estrogen receptors was found in the cytosols and nuclear extracts of the adrenal cortex proper and special zone of the brushtail possum (Trichosurus vulpecula). 2. The specific estradiol receptor had a sedimentation coefficient on sucrose gradients of approximately 9S and a molecular weight on gel filtration of more than 200,000. The adrenal cortex cytosol binds [3H]estradiol with high affinity (Ka 5.5 X 10(9) M-1), and limited capacity (Bmax 62.7 fmol/mg cytosol prot). In competition experiments with different steroids the receptor showed a high affinity for four estrogens and a very low affinity to androgens, progesterone and cortisol. 3. There was no difference in the affinity and maximum binding capacity of the cytosols from cortex proper in male and female animals, but the binding capacity of the special zone of females was half that of cortex proper. Estradiol receptors were found in the kidney, liver, lung, testis and muscle but only in the adrenal and prostate was the binding capacity relatively high compared with the uterus. 4. The specific binding capacity of [3H]estradiol to cytosols of adrenal cortex at different stages of the estrus cycle and pregnancy was unrelated to that of the uterus. In the adrenal the receptor concentration was lowest at estrus, when uterine concentration was high, while in late pregnancy the binding of adrenal cortex and uterus cytosols was almost the same. 5. The possible physiological significance of the presence of a specific estrogen receptor in male and female possums is discussed.
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Previous studies have suggested that basic fibroblast growth factor (bFGF) is involved in the mediation of the compensatory adrenal growth response. These studies were undertaken to identify the bFGF receptor in the rat adrenal cortex and determine bFGF receptor levels in vivo. Initial studies using primary cultures of rat glomerulosa cells demonstrated a high affinity binding site with a Kd of 10 pM, consistent with reported values for the bFGF receptor; however, these results could not be demonstrated in capsule-glomerulosa tissue. Using autoradiography to verify the existence of bFGF receptors in vivo, heparin-insensitive [125I]bFGF-binding sites were concentrated primarily in the capsule and glomerulosa of the adrenal cortex, suggesting the presence of FGF receptors. The presence of bFGF receptors was further verified by demonstration of internalization of [125I]bFGF into cells of capsule-glomerulosa preps. This approach was used to demonstrate that suramin (a bFGF antagonist) pretreatment of rats results in bFGF receptor up-regulation in the adrenal cortex. In addition, we demonstrated a decrease in bFGF internalization in the remaining adrenal 24 h after unilateral adrenalectomy, suggesting the utilization of bFGF during the compensatory adrenal growth response. Together, these data support a role for bFGF in autocrine stimulation of the adrenal cortex and in the compensatory adrenal growth response.
The adrenal cortex function of farmed blue (Alopex lagopus) and silver foxes (Vulpes vulpes) differing in their reaction in the feeding test were assessed. The urine cortisol:creatinine ratio was lower for those animals eating in the feeding test in comparison to those not eating in both species. In addition, eater silver foxes had lower baseline serum cortisol concentration and also lower serum cortisol concentration 2 h after ACTH administration than noneaters. There were no differences in any serum cortisol levels between the eater and noneater blue foxes. The weights of body and adrenals did not differ between confident and fearful animals in either species. The present study demonstrates that animals not eating in the feeding test may have higher fearfulness and be more stressed than animals eating.
The study aimed to examine the effects of pinealectomy (PX) and melatonin on adrenal cortex of orchiectomized rats and to explain whether the testosterone action on adrenal cortex is modified by the pineal gland. Neither PX (53 days) nor melatonin administration (10 days) had an effect on the structure as well as on the studied functional parameters of the adrenal gland--corticosterone output by homogenates, 11 beta-hydroxylase and 5 alpha-reductase activities, liver corticosterone metabolism and serum corticosterone level. On the contrary, PX of 14 months duration resulted in an increase in 5 alpha-reductase activity. Testosterone administration into orchiectomized rats lowered corticosterone output and liver corticosterone metabolism and these effects were unaffected by PX or melatonin. Testosterone lowered 5 alpha-reductase and 11 beta-hydroxylase activities, the effects not modified by PX, however, after melatonin administration activities of these enzymes were similar as in orchiectomized rats. Obtained results cast doubts on a physiological role of the pineal gland and its principal hormone--melatonin--in regulation of adrenocortical structure and secretory activity and suggest that testosterone action on the rat adrenal cortex is not modified by the pineal gland.
Mononuclear phagocytes of the Rat adrenal cortex were identified by light microscopy after injection of Chinese ink or colloidal iron (Imferon 200). They may be found as endothelial, periendothelial or intracapillary cells. The number of mononuclear phagocytes was much greater in the adult than in the young Rat, with a significant peak 3 hrs. after injection of Chinese ink in the young Rat and two peaks, 4 and 12 hrs. after injection in the adult.
At autopsy there was found extra-adrenal pheochromocytoma, bilateral diffuse-nodulous and adenomatous hyperplasia of the adrenal cortex in a female patient who had suffered for a long time from high arterial pressure. In the enlarged thyroid gland there was revealed micro-macrofollicular colloid goiter with basedowification. The morphological changes and results of an investigation of the liquor (drop in the content of potassium) justify the assumption that in this case there is a combination of extra-adrenal pheochromocytoma with hormonally active rearrangement of the adrenal cortex--primary hyperaldosteronism and basedowified goiter.
Morphological investigation of the rat adrenal cortex and corticosterone contents in the blood plasma demonstrated certain peculiarities in the adrenal reaction to the primary and repeated immobilization stress in intact animals and after hypokinesia. It has been stated that morphological manifestation to an acute stress reaction kinesia is less pronounced than at an analogous single effect. Investigation on corticosterone content in blood plasma revealed certain functional peculiarities in the adrenal cortex reactions in different variants of the experiments. Five hours after the beginning of the stress, corticosterone content was increased only at the initial immobilization, while at repeated stimulations during the same time it did not differ from the control. Changes in functional possibilities of the adrenal cortex in response to a repeated stimulation and in response to a new stimulation in animals previously adapted to another type of stress are discussed.
Two cases of adenomas of adrenal cortex were examined. Both cases presented clinical symptoms of autonomic hyperaldosteronism. Histologically one adenoma had a benign structure, the other a malignant one. An ultrastructural and cytochemical assay of the benign form suggests that it is composed of so called "hybrid cells". Histochemically the cells of both adenomas were characterized by a 3-beta hydroxysteroid and succinic dehydrogenase reaction, the presence of MAO and nonspecific esterases and in some single cells 17-beta hydroxysteroid dehydrogenase.
Several analogs of angiotensin in which the phenylalanine in position 8 of the peptide chain was replaced by an aliphatic amino acid residue are specific antagonists of angiotensin in aorta, the adrenal medulla, and adrenal zona glomerulosa. In the adrenal cortex and medulla, all actapeptide analogs have more agonist activity than in aortic strips. In studies with N-terminally substituted analogs, it appears that adrenal degradation of the angiotensin molecule by aminopeptidase(s) does not occur or is not retarded by N-terminal mocifications such as sarcosine substitution. The decapeptide analog [Ile8]-angiotensin I and heptapeptide analog [des-Asp1, Ile8]-angiotensin II were excellent antagonists in the adrenal medulla and each peptide was devoid of intrinsic activity. These substituted homologs of angiotensin may offer a novel approach for the development of selective antagonists of angiotensin receptors. In the adrenal cotex, [des-Asp1, Ile8]-heptapeptide possessed greater receptor affinity than any of the angiotensin octapeptides studied. This C-terminally substituted heptapeptide does have significant intrinsic activity in the adrenal cortex which would limit the use of this compound as an antagonist of vascular responses to angiotensin II. In studies with [Ile8]-angiotensin II, [Sar1, Ile8]-angiotensin II, and [des-Asp1, Ile8]-angiotensin II, the pA2 values calculated indicate that the N-terminal residue is not important for receptor binding in the adrenal cortex but may be of significance in binding to adrenal medullary and aortic smooth muscle receptors. At the present time it appears unlikely that any single animal model or assay system can reliably predict the agoinst/antagonist activities of angiotensin analogs for all the various end organs which respond to the angiotensins.
1. Mitochondria isolated from porcine adrenal cortex under State 3 conditions oxidized succinate with a rate of 47 +/- 4.48 na oxygen/min/mg/protein and with ADP:O ratio 0.98 +/- 0.09. In the presence of 15 microM deoxycorticosterone the rate of succinate oxidation was 36.8 +/- 3.08 na oxygen/min/mg/protein. 2. Under the same conditions the rate of glutamate oxidation was 22.8 +/- 2.21 and 16.8 +/- 0.65 na oxygen/min/mg/protein, respectively. ADP:O ratio was 1.45 +/- 0.14. 3. Introduction of trace amounts of malate into the mitochondria oxidizing glutamate only slightly increased the rate of O2 uptake. 4. The glutamate dehydrogenase activity in these mitochondria was 12.5 +/- 0.69 nmol/min/mg.
Particulate guanylate cyclase from bovine adrenal cortex can be stimulated by ANF. A 2-fold stimulation of the enzyme was obtained with 100 nM ANF and a half-maximal stimulation, with a 5 nM dose. The stimulation by ANF persisted for at least 30 min. Various detergents, such as Triton X-100, Lubrol PX, cholate, CHAPS, digitonin and zwittergent, stimulated several-fold the activity of particulate guanylate cyclase. However, only Triton X-100 dispersed particulate guanylate cyclase without affecting its response to ANF. The dose-response curve of ANF stimulation of the particulate and the Triton X-100 dispersed enzyme was similar. The dispersion of a fully responsive guanylate cyclase to ANF will help us to uncover the type of interactions between guanylate cyclase and ANF. It will also be used as a first step for the purification of an ANF-sensitive particulate guanylate cyclase.
Zona fasciculata cells from the adrenal cortex of female Sprague-Dawley rats were fixed by immersion in potassium pyroantimonate-osmium tetroxide and potassium pyroantimonate-glutaraldehyde to study the distribution of calcium. Potassium pyroantimonate-osmium tetroxide treatment gave reproducible patterns of electron-opaque precipitate, whereas inconsistent deposits of reaction product were seen after potassium pyroantimonate-glutaraldehyde fixation. Nuclei showed sparse precipitate over heterochromatin and dense aggregates over areas of nucleoli surrounded by portions of the nucleolar-dense component. Two major cytoplasmic sites of precipitate were identified: mitochondria and vesicles of smooth endoplasmic reticulum. Most of the intramitochondrial precipitate was localized to the intracristal space. Precipitate was also seen in vesicles of Golgi apparatus. The extracellular space was filled with closely packed electron-opaque particles. Observation of tissues treated with control fixative saturated with EGTA showed little if any reaction, confirming that calcium was the primary cation precipitated by potassium pyroantimonate. Our results provide a method suitable for accurate localization of calcium in adrenocortical cells.
Malate dehydrogenase from bovine adrenal cortex has been purified to homogeneity, using affinity chromatography on 2',5'-ADP-Sepharose 4B. The kinetic data do not contradict the consecutive mechanism of the reaction with the ordered addition of substrates: NADP binds first, then malate. The enzyme conformation initiated by NADP and malate binding is less thermostable. Malate dehydrogenase has intrinsic tryptophan fluorescence with the spectrum maximum at 335 +/- 1 nm, half-width of 50 +/- 1 nm and quantum yield of 0.08. The tryptophan residues belonging to class 1 (75%) and class 2 (25%) make the main contribution to the intrinsic fluorescence of malate. The binding of cofactors and substrates results in the quenching of enzyme fluorescence. The values of dissociation constants for malate dehydrogenase complexes with NADP (4 microM), with NADP . H (8 microM) and with pyruvate (2.5 mM) correlate with the corresponding values of Km. The shifts in pH of the medium induce changes in the fluorescence parameters which are probably related to conformational changes in the enzyme molecule. The changes in the fluorescence parameters correlate with the alterations of the malate dehydrogenase enzymatic activity.
A case of Sweet's syndrome preceding carcinoma of the adrenal cortex is reported. Sweet's syndrome can be a rare premalignant manifestation of carcinoma of adrenal cortex.
The role of the pituitary in the regeneration of the adrenal cortex was examined. Enucleation of the left adrenal, and removal of the right adrenal were performed in hypophysectomized rats, in sham-operated control rats and in intact control rats. The mitotic activity of adrenal regenerates was evaluated on the 5th day after enucleation. A statistically significant decrease (p less than 0.001) in the mean mitotic activity rate was noted in the hypophysectomized rats, when compared with both control groups. The results obtained indicate a significant stimulating effect of the pituitary on the regeneration of the adrenal cortex in rats.