Clinical chronopharmacology of ACTH 1-17. I. Effects on plasma cortisol and urinary 17-hydroxycorticosteroids

The aim of the investigation was to study the effects of ACTH 1-17 on both plasma cortisol and urinary 17-OHCS in health adult young males with regard to the time (clock hours) at which this polypeptide was injected. Eight healthy adults (males from 18-30 years) volunteered for the study. They were synchronized with a diurnal activity from 0700 to 0000 and a nocturnal rest. Each week, during 6 consecutive weeks (January 19 to February 25, 1980), a 3-day test was performed on Saturday, Sunday and Monday. On Sundays 3 control-tests and 3 ACTH-tests were programmed during which either saline or 100 micrograms ACTH 1-17 were injected i.m. at respectively 0700, 1400 and 2100. During each 3 day-test (72 h) the urinary excretion of 17-OHCS was determined every 4 h at fixed clock hours. In addition, on Sundays, venous blood was sampled prior to control or ACTH injections at respectively 0700, 1400, and 2100 and 20, 40, 60, 90, 120, 150 and 180 min thereafter. Plasma cortisol (radioimmunoassay) was determined in samples thus collected. Both conventional and cosinor methods were used for statistical analyses. A strong and statistically significant rise of plasma cortisol was observed after all of the ACTH 1-17 injections. The obtained mean response curves were observed after all of the ACTH 1-17 injections. The obtained mean response curves were similar in form and parallel. The highest plasma cortisol curve corresponded to ACTH injected at 0700, the lowest to ACTH injected at 2100. The curve corresponding to ACTH injected at 1400 went in-between. The 24-h urinary excretion of 17-OHCS after ACTH 1-17 was approximately 4 times greater than the control value when injected at 0700, approximately 3 times greater than control when injected at 1400 and only twice greater than control when injected at 2100. In terms of changes in plasma cortisol and 17-OHCS the greatest best benefit of ACTH 1-17 is achieved when this polypeptide is injected at 0700, rather than at 1400 or 2100 in diurnally active subjects.     

Effect of glucose on adrenocortical activity of newborn piglets.

Adrenocortical function was followed in newborn piglets fasted for 36 hours and in piglets given only glucose or glucoplastic amino acids during this period. The greatest increase in relative adrenal weight, blood plasma 17-hydroxycorticosteroid (17-OHCS) levels and in the production of 17-OHCS by pig adrenals in vitro as compared with suckled controls was found in fasted piglets followed by amino acid-treated animals. The latter showed no significant differences in blood glucose between the initial and final values. The administration of glucose produced a transient hyperglycemia, failed to prevent the inhibition of body growth, resulted in a reduction in liver weight similar to that seen in fasted animals, and had a marked suppressive effect on the elevation of adrenocortical function. There were no significant differences in the production of 17-OHCS by adrenals in vitro per kg body weight between the glucose-treated and suckled piglets.     

Effect of Resection of Lung Tumours on the Steroid Abnormalities in Patients with Lung Cancer

The urinary excretion of androsterone, aetiocholanolone, total 17-oxosteroids, and 17-hydroxycorticosteroids (17-OHCS) was measured in 40 patients with lung cancer three days before resection and again 10-15 days after resection of their lung tumours. There was a significant postoperative increase in the excretion of 17-OHCS but a significant decrease in the excretion of androsterone and aetiocholanolone, resulting in an increase of the preoperative abnormalities in steroid excretion in these patients. Since there was no change in steroid excretion towards normal after resection of the lung tumours, it seems that the steroid abnormalities found in lung cancer are not the effect of the presence of the lung tumours. As the excretions of 17-OHCS and 11-deoxy-17-oxosteroids change in opposite directions after resection, it is suggested that a dissociation of factors that control the excretion of these two groups of steroids takes place as a response to surgical stress in patients with lung cancer.     

Progesterone secretion by adrenal glands of hamsters and comparison of ACTH influence in rats and hamsters.

Studies were designed to determine a) if adrenal glands of hamsters secrete progesterone (PROG), b) the effects of adrenocritocotropin (ACTH) administration on adrenocortial function of rats and hamsters under the surgical conditions necessary for collection of adrenal venous blood from the left renal vein, and c) the effects of blood loss during sample collection. PROG was quantitated by the competitive protein-binding method after extraction and separation by sephadex LH-20 column chromatography. The presence of interfering quantities of androstenedione necessitated two column chromatographic steps. Glucocorticoids (11-OHCS) were determined fluorometrically. PROG was detected in adrenal venous plasma of female hamsters. The PROG concentration and secretory rate were 91 +/- 12 ng/ml and 4 +/- 1 ng/min, respectively, while the peripheral plasma level of the same animals was 2 +/- 0.2 ng/ml, indicating that the adrenal glands of female hamsters are capable of secreting PROG. ACTH administration increased PROG secretory rates in both hamsters (3 +/- 1 to 14 +/- 3 ng/min) and rats (62 +/- 9 to 152 +/- 32 ng/min) on estrus, as well as increasing the 11-OHCS secretory rate of hamsters (16 +/- 1 to 33 +/- 4 ng/min), but not of rats. The greater increase in PRCC than in 11-OHCS secretion may be related to excess PROG formation relative to the capacity of the 17alpha- or 21-hydroxylating enzyme systems. The adrenal venous PROG concentration and secretory rate of female hamsters infused with 10% dextran while collecting adrenal venous blood did not differ significantly from those of the non-infused animals, suggesting that this amount of blood loss (1 ml) does not influence PROG secretion.     

Effect of an oral contraceptive on the lysine-8-vasopressin test in the diagnostic evaluation of pituitary function.

A comparative study was performed on the plasma 11-hydroxycorticosteroids (11-OHCS) responses to lysine-8-vasopressin (LVP) before and during ingestion of an oral contraceptive wieh oestrogenic activity. A total of 19 healthy women with a normal menstrual cycle and normal plasma 11-OHCS content in blood samples at 8 a.m. and 4 p.m. were included in the study. The results show that the mean response of plasma 11-OHCS to the administration of LVP was equal in magnitude before and during ingestion of the contraceptive, although the baseline value of 11-OHCS doubled as a result of the treatment. On the other hand, during contraceptive treatment the increase of plasma 11-OHCS concentration after the administration of LVP was somewhat slower, reaching its peak later than before treatment. The noticeable variation in individual responses of plasma 11-OHCS to LVP increased during the treatment. Three subjects showed weak responses to LVP, although the adrenocortical responses to exogenous ACTH were quite normal. The inidvidual responses to LVP before and during ingestion of an oral contraceptive with oestrogenic activity are discussed.     

Changes of maternal and fetal pituitary-adrenocortical functions during human labour

The results of maternal 11-hydroxycorticoid (11-OHCS) determination on blood obtained from four stages of 44 spontaneous labour cases showed a strong and time-related maternal adrenocortical activation during vaginal delivery. The high 11-OHCS level in the cord blood and the increase of the 11-OHCS concentration in the amniotic fluid suggested the same response of the fetal adrenal. However, in 14 cases of elective caesarean sections the 11-OHCS levels in the cord blood and the amniotic fluid did not rise in spite of the elevated maternal plasma 11-OHCS content.     

A case report of Turner's syndrome with ring X chromosome.

A case of Turner's syndrome with short stature and 45, XO/46, XXr mosaicism in chromosome study was presented. With special emphasis on endocrinological study, the size of the breasts was normal in contrast to the poor development of the breasts in most of types of Turner's syndrome. She showed normal thyroid function, slightly low level of urinary 17-OHCS, decreased 17-KS, poor response in metopirone test and poor response of HGH to insulin.     

Inverse correlation between urinary 17-hydroxycorticosteroid (17-OHCS) excretion and thyrotropin-releasing hormone (TRH)-induced thyrotropin (TSH) response in disorders of adrenocorticotropic hormone (ACTH) and adrenal secretion

A TRH test was performed in patients with Cushing's syndrome and isolated ACTH deficiency, and serum TSH was measured. A TRH-induced TSH showed low response in patients with Cushing's syndrome but showed excessive response in patients with isolated ACTH deficiency. However, in both diseases TRH-induced TSH response showed a tendency to normalize after the treatment. Urinary 17-OHCS excretion was measured in patients with Cushing's syndrome and isolated ACTH deficiency before and after the treatment. There was a definite inverse correlation between ;the logarithm for urinary 17-OHCS excretion and the peak TSH response to TRH. These results suggested that glucocorticoids regulate not only ACTH but also TSH secretion.     

Preparation of electrophoretically homogeneous erythrocuprein and its thermodenaturation]

The procedure for isolation of electrophoretically homogeneous preparation of erythrocuprein from 17 1 blood batch was described. The preparation had absorbtion value, A260/A680, of 25-26. The protein completely retains superoxide dismutase activity after treatment 7 min. at 75 degrees C. The heating of protein solution at 100 degrees C bring about autoreduction of protein copper as demonstrated by decreasing optical and EPR-spectral intensities. The reduced protein was oxidized in aerobic condition and oxidized preparation had optical and EPR-spectra differ from those of the native protein the activy being decreased.     

Assessment of Long-acting Synthetic Corticotrophin in Hypersensitive Asthmatics and Normal Subjects

The synthetic polypeptide depot-tetracosactrin (Synacthen-Depot) was given to nine steroid-treated asthmatic patients hypersensitive to animal corticotrophin. Eight had a satisfactory increase in plasma and urinary 17-hydroxycorticosteroids (17-OHCS). The remaining patient had been shown previously to have adrenal suppression. None of the patients developed allergic reactions to depot-tetracosactrin, and seven have been receiving regular injections of the polypeptide twice weekly for the past eight months.In 13 normal subjects intramuscular injections of 1 mg. and 0.5 mg. of depot-tetracosactrin and 40 units of corticotrophin-gel were equally potent in raising the plasma 17-OHCS, but depot-tetracosactrin had a significantly longer action than corticotrophin-gel.     

Hypertension,Increased Aldosterone Secretion and Low Plasma Renin Activity Relieved by Dexamethasone

A father and son are described with a condition characterized by benign hypertension, potassium deficiency, increased aldosterone secretion rate (ASR), raised plasma volume and suppressed plasma renin activity (PRA). There were intermittent elevations of urine 17-ketosteroids and 17-hydroxycorticoids (17-OHCS) but no increase in urine THS, normal circadian rhythm of plasma 17-OHCS, and normal urine 17-OHCS response to dexamethasone and intravenous ACTH. Plasma ACTH and corticosterone secretion were not elevated. Pregnanetriol excretion was normal but urine pregnanediol was increased. At operation on the father no adrenal tumour was found; the excised left adrenal weighed 7 g. and showed nodular cortical hyperplasia; juxtaglomerular cells showed only occasional granules. Following operation hypertension persisted and ASR was half the preoperative value. All abnormalities in father and son were relieved by dexamethasone (DM) 2 mg. daily. The condition recurred following cessation of DM but was relieved by a second course of treatment. No such response to DM was seen in a normal subject or in a patient with Conn''s syndrome. For a number of reasons it is suggested that patients with hypertension, increased ASR and low PRA be given a trial of dexamethasone treatment before undergoing adrenal surgery.     

Circadian rhythms of electrolyte and 17-hydroxycorticosteroid excretion in hyperthyroidism and hypothyroidism.

The circadian rhythms of excretion of sodium, potassium, calcium, magnesium, phosphorus and 17-hydroxycorticosteroid (17-ohcs) were determined in five normal subjects, in six patients with hyperthyroidism and five with hypothyroidism. Constant diets with identical 3-hourly feedings were employed, and urine collections were made every 3 hrs during a 3-day study period. The circadian patterns of urinary excretion of sodium, potassium and 17-OHCS were similar in all three groups with distinct daytime peaks and nighttime nadirs. The total quantities of the ions and 17-OHCS excreted were greater in hyperthyroid than in hypothyroid patients with the greatest difference noted with the 17-OHCS. The rhythms for calcium, magnesium and phosphorus excretion were accentuated in hyperthyroid patients but similar to those in normal subjects with early morning calcium and magnesium peaks and a phosphorus peak approximately 12 hrs later. While a similar although blunted circadian pattern for calcium and perhaps magnesium excretion was noted in hypothyroid patients, their phosphorus rhythms were distorted and rather flat. These latter results confirm the observation of MINTZ et al. and are compatible with their interpretation that thyroid hormone is permissibly necessary for the expression of a normal phosphaturic rhythm and that the circulating level of thyroid hormone influences the amplitude of the phosphaturic rhythm.     

A prospective study of corticosteroid and catecholamine levels in relation to viral respiratory illness.

Urinary 17-OHCS, epinephrine and norepinephrine levels were studied for one week before and one week after the onset of acute, severe Adenovirus 4 respiratory illness in 12 Army recruits during Basic Combat Training. During the pre-illness period, a tendency was frequently noted for all three hormone levels to show "spiking" elevations two to four days before illness onset. There was also a tendency for 17-OHCS levels to rise on the day before fever onset. The possible relationship of these pre-illness hormonal changes to stressful experiences and, in turn, to altered host resistance to infectious illness is discussed. Following onset of respiratory illness, 17-OHCS, epinephrine and norepinephrine levels all showed about 60 percent increases over the early pre-illness period baseline value. Elevations of these hormones persisted for about four to five days, roughly in correlation with fever duration, with only slight differences in configuration and timing of curves from one hormone to the next. The problem of evaluating which of several independent variables operating concurrently during infectious illness may be responsible for stimulating the final common neuroendocrine pathways is discussed.     

Effect of bone injury on circadian periodicity of plasma 17-hydroxy corticosteroids (17-OHCS) in rabbits

Bone injury inflicted at varying time intervals during 24 hr day-night cycle caused significant varying increase in plasma 17-OHCS levels in all traumatized animals and the levels remained elevated up to 24 hr after trauma. The level of plasma 17-OHCS was found to be aberrated in all the traumatized animals. Thus, adequate adrenocortical response to trauma and aberration in the adrenocortical secretory activity appears inevitable irrespective of the time at which the trauma is produced. However, the degree of response depends on the particular time at which the trauma is inflicted.     

Licorice reduces serum testosterone in healthy women

Licorice has been considered a medicinal plant for thousands of years. The most common side effect is hypokalemic hypertension, which is secondary to a block of 11beta-hydroxysteroid dehydrogenase type 2 at the level of the kidney, leading to an enhanced mineralocorticoid effect of cortisol. We have investigated the effect of licorice on androgen metabolism in nine healthy women 22-26 years old, in the luteal phase of the cycle. They were given 3.5 g of a commercial preparation of licorice (containing 7.6% W.W. of glycyrrhizic acid) daily for two cycles. They were not on any other treatment. Plasma renin activity, serum adrenal and gonadal androgens, aldosterone, and cortisol were measured by radioimmunoassay. Total serum testosterone decreased from 27.8+/-8.2 to 19.0+/-9.4 in the first month and to 17.5+/-6.4 ng/dL in the second month of therapy (p<0.05). It returned to pre-treatment levels after discontinuation. Androstenedione, 17OH-progesterone, and LH levels did not change significantly during treatment. Plasma renin activity and aldosterone were depressed during therapy, while blood pressure and cortisol remained unchanged. CONCLUSIONS: Licorice can reduce serum testosterone probably due to the block of 17-hydroxysteroid dehydrogenase and 17-20 lyase. Licorice could be considered an adjuvant therapy of hirsutism and polycystic ovary syndrome.     

Adrenal function in different phases of the estrous cycle in the domesticated silver fox, Vulpes fulvus

Experiments were made on silver foxes from a population which had been selected for 10 to 15 generations for the domestic behaviour and on animals from a control, unselected population. In females from both populations, studies were made of the level of 11-OHCS in the blood serum, in vitro production of 11-OHCS, the size of fascicular zone in the adrenal cortex, the volume of cellular nuclei and nucleoli, as well as the reaction of the adrenals in vitro to 2 doses of ACTH (1 and 5 units/g of the adrenals) during anestrus, proestrus and estrus. In control females, all the characters investigated significantly increased from anestrus to proestrus. In domesticated females, no changes in the production of 11-OHCS in vitro or changes in morphological features of the fascicular zone were observed in the course of estrous cycle. During proestrus, the adrenals of the domesticated animals were not able to increase the production of 11-OHCS in vitro after application of ACTH. The decrease in the reactivity of the adrenals to the effect of ACTH is presumably the main cause why in unselected females the adrenal are not activated during proestrus. Therefore, in the course of selection for the domestication type of behaviour, species specific dynamics of activation of the adrenals during estrous cycle is lost.     

Size changes of mouse ova during preparation for morphometric studies in the electron microscope

Summary The diameters of oocytes during successive steps of preparation for electron microscopy were measured in the light microscope. During glutaraldehyde fixation the diameter of the ova mainly reflected the osmolarity of the fixative vehicle. An increase in the diameter of the ova occurred with postfixation in OsO₄ depending on time of postfixation and also on osmolarity and OsO₄ concentration. During ethanol dehydration the cell diameter decreased to its lowest value and remained unchanged after embedment in Epon. A procedure that minimizes ova size changes during preparation for electron microscopy is elaborated.This investigation was supported by NIH PL 480 Research Agreement No. 05-030-1.     

Change in the absorptive capacity of macrophages from different organs after administration of hydrocortisone

Two, twenty-four and 48 h after hydrocortisone treatment in a dose of 125 mg/kg bw the blood clearance rate for colloidal carbon particles in rats turned to be 2, 2.1. and 1.6 times less whereas that for 51Cr-SRBC in CBA mice 2.1, 2.2 and 1.7 times less as compared to untreated controls. Within 24 and 72 h after hormone injection the efficacy of red blood cell uptake by Kupffer cells decreased 1.35 and 1.8 times whereas the similar uptake by lung or spleen macrophages changed but insignificantly and that by bone marrow cells was even greater than in controls. Toward the 5th day after zymosan treatment the uptake capacity of Kupffer cells was the greatest whereas the plasma 11-OHCS content was 1.3-fold less versus the control values.     

游泳对大鼠阳虚状态的影响及数理分析

目的考察一次性游泳对氢化可的松粉针剂诱导大鼠阳虚状态指标的影响,以期暴露模型本身内隐的隐性测评指标并确定特征指标。方法将SD雄性大鼠随机分为正常组、正常游泳组、阳虚组、阳虚游泳组,阳虚组／阳虚游泳组肌注氢化可的松粉针剂20ms／kg,正常组／正常游泳组注射等量的0．9％氯化钠溶液,注射14d后,阳虚游泳组／正常游泳组辅以一次性游泳后,各组麻醉,取血,测定相关指标。结果阳虚游泳组C4、cGMP、TC、L阳虚隐性指标暴露,BUN、17-OHCS、T、T／E2、cAMP／cGMP阳虚评定指标凸显。结论非破坏性一次游泳可以使氢化可的松粉针剂诱导的大鼠阳虚证候模型的隐性指标凸显、部分显性指标放大,建议今后在建立、模拟阳虚证候模型时辅以游泳手段,以便客观、全面、真实的反映模型状态。     

Aminoglutethimide effect on circadian rhythms in urinary variables in a patient with idiopathic hyperaldosteronism

Aminoglutethimide (AG: 750 mg/day) was administered to a patient with idiopathic hyperaldosteronism (IHA) and circadian rhythms in urinary excretion of sodium (UNaV), potassium (UKV), aldosterone (AER) and 17-OHCS were analyzed by the single cosinor method. Urine was collected every 4h for 24h on the day before and on the 1st, 3rd and 7th day of AG administration, and above variables in each sample were determined. Circadian rhythms of 14 patients with primary aldosteronism (PA) who served as controls were also analyzed. In the present case, circadian acrophases in UNaV and AER studied before AG administration occurred at 22(19) and 07(05), respectively. They were similar to those of preoperative PA-patients. Circadian acrophase in UNaV occurred earlier with AG administration and on the 7th day it was at 14(05), a value similar to that of postoperative PA-patients. Circadian mesor in AER decreased remarkably from 4.1 to 0.6 micrograms/4h with AG administration, as did circadian mesor in UKV, whereas circadian mesor and acrophase in 17-OHCS did not change. Thus, the circadian characteristics in urinary variables in the present IHA-case were pathophysiologically similar to those of PA.