Comparison of different enzyme-immunoassays for assessment of adrenocortical activity in primates based on fecal analysis.

Most studies published to date that used fecal glucocorticoid measurements to assess adrenocortical activity in primate (and many nonprimate) species applied a specific cortisol or corticosterone assay. However, since these native glucocorticoids are virtually absent in the feces of most vertebrates, including primates, the validity of this approach has recently been questioned. Therefore, the overall aim of the present study was to assess the validity of four enzyme-immunoassays (EIAs) using antibodies raised against cortisol, corticosterone, and reduced cortisol metabolites (two group-specific antibodies) for assessing adrenocortical activity using fecal glucocorticoid metabolite (GCM) measurements in selected primate species (marmoset, long-tailed macaque, Barbary macaque, chimpanzee, and gorilla). Using physiological stimulation of the hypothalamo-pituitary-adrenocortical (HPA) axis by administering exogenous ACTH or anesthesia, we demonstrated that at least two assays detected the predicted increase in fecal GCM levels in response to treatment in each species. However, the magnitude of response varied between assays and species, and no one assay was applicable to all species. While the corticosterone assay generally was of only limited suitability for assessing glucocorticoid output, the specific cortisol assay was valuable for those species that (according to high-performance liquid chromatography (HPLC) analysis data) excreted clearly detectable amounts of authentic cortisol into the feces. In contrast, in species in which cortisol was virtually absent in the feces, group-specific assays provided a much stronger signal, and these assays also performed well in the other primate species tested (except the marmoset). Collectively, the data suggest that the reliability of a given fecal glucocorticoid assay in reflecting activity of the HPA axis in primates clearly depends on the species in question. Although to date there is no single assay system that can be used successfully across species, our data suggest that group-specific assays have a high potential for cross-species application. Nevertheless, regardless of which GC antibody is chosen, our study clearly reinforces the necessity of appropriately validating the respective assay system before it is used.

[Effect of striatum destruction on the hypothalamo-pituitary-adrenal axis function in adrenalectomized rats]. / Vliianie povrezhdeniia striatuma ha funktsiiu gipotalamo-gipofizarno-nadpochechnikovoi sistemy u adrenaléktomirovannykh krys.

The influence of the bilateral striatectomy on activation of the hypothalamus-pituitary-adrenal axis after unilateral adrenalectomy was investigated. Increasing of the intact adrenal gland's mass in striatectomized rats was less than in the control group. It was combined with decreasing of the production of corticosteroids by intact gland. Such disturbances were more obvious in rats with destruction of the dorsal striatum than the ventral one. These facts suggest the striatum participation, its dorsal zone in particular, in the processes of regulation of the hypothalamus-pituitary-adrenal axis.

The presence of biologically significant concentrations of glucocorticoids but little or no cortisol binding globulin within aqueous humor: relevance to immune privilege in the anterior chamber of the eye.

PURPOSE: Immunosuppressive factors in aqueous humor (AH) contribute to the immune-privileged status of the anterior chamber of the eye. One such factor is transforming growth factor-beta (TGF-beta); other relevant inhibitors have not been fully identified. The authors examined AH to search for other putative inhibitors and to determine their effect on TGF-beta inhibitory activity. METHODS: Radioimmunoassays (RIA) were used to detect the presence of hydrocortisone, corticosterone, cortisol binding globulin (CBG), and alpha-melanocyte stimulating hormone (alpha-MSH) in AH. The ability of these factors to inhibit murine thymocyte proliferation stimulated by phytohemagglutinin-interleukin 1 (PHA/IL-1) and proliferation of a TGF-beta-sensitive cell line (CCL64) in vitro was examined. The ability of hydrocortisone to inhibit a one-way mixed lymphocyte reaction (MLR) and the ability of epidermal cells to present soluble tumor-associated antigens (TAA) for elicitation of immunity in mice in the concentration range present in AH was also examined. RESULTS: Hydrocortisone was detected in mouse, rat, and human AH (10.8 +/- 1.1 ng/ml, 9.3 +/- 2.1 ng/ml, and 18.0 +/- 1.0 ng/ml, respectively; mean +/- SEM), as was corticosterone (2.7 +/- 0.9 ng/ml, 2.2 +/- 0.3 ng/ml, and 0.7 +/- 0.1 ng/ml, respectively). Whereas normal plasma contains a binding protein for corticosteroids (i.e., CBG), the concentration in mouse, rat, and human AH was less than the level detectable by an RIA. Hydrocortisone inhibited PHA/IL-1-stimulated murine thymocyte proliferation and CCL64 cell proliferation in the concentration range present in AH. When hydrocortisone was combined with TGF-beta 2 (125 pg/ml), the degree of inhibition observed was greater than with either alone. Corticosterone inhibited thymocyte costimulation only slightly at concentrations present in AH but was inhibitory for CCL64 cells. alpha-MSH was also detected in AH. The concentration present had only slight inhibitory effects for CCL64 cell proliferation and did not enhance TGF-beta 2-mediated (62 pg/ml to 250 pg/ml) inhibition of CCL64 or thymocyte proliferation. Hydrocortisone inhibited the one-way MLR in the concentration range present in AH and, at 10 ng/ml, inhibited the ability of epidermal cells to present TAA for elicitation of delayed-type hypersensitivity in tumor-immune mice. CONCLUSIONS: These results show that AH contains biologically relevant concentrations of glucocorticoids and that CBG is relatively absent so that glucocorticoids present are largely free, and they suggest that regional sites take advantage of the activities of multiple factors to maintain an immune-privileged status.

[The reaction of the hypothalamo-hypophyseal-adrenal cortex system in rats to unilateral adrenalectomy in hypoparathyroidism]. / Reaktsiia gipotalamo-gipofizarno-adrenokortikal'noi sistemy krys na odnostoronniuiu adrenaléktomiiu pri gipoparatireoze.

Parathyroidectomy and subsequent hypocalcemia were shown to inhibit the response of the hypothalamo-hypophyseal-adrenocortical system (HHACS) to unilateral adrenalectomy, as expressed by a lesser compensatory hypertrophy of intact adrenal gland and a lower concentration of 11-oxycorticosteroids in adrenal gland and in plasma. The greatest activity of the HHACS was revealed in 3-month old rats. The data obtained suggest an interaction between parathyroid and adrenal glands.

[The direct stimulating effect of arginine vasotocin on the functional activity of the interrenal gland in the frog Rana temporaria]. / Priamoe stimuliruiushchee vliianie arginin-vazototsina na funktsional'nuiu aktivnost' interrenalovoi zhelezy liagushki Rana temporaria.

Using radioimmunoassay it has been detected that both nonoperated and hypophysectomized, lacking endogenous ACTH, frogs injected one or three times with arginine vasotocin (5.10(-9) M/kg b. w.) show a statistically significant increase of plasma corticosterone level as compared with that in control animals and frogs injected with Ringer solution. The level of 11-hydroxycorticosteroids (fluorometric determination) in the interrenal gland decreases significantly only in animals three times injected with arginine vasotocin. It is assumed that arginine vasotocin produces a direct stimulatory effect on corticosteroid-producing cells of the frog interrenal gland.

[The circadian rhythm of the adrenal glucocorticoid function in female rats selected for the reaction of the estrous cycle to constant illumination]. / Sutochnyi ritm gliukokortikoidnoi funktsii nadpochechnikov samok krys, selektsionirovannykh po reaktsii éstral'nogo tsikla k postoiannomu osveshcheniiu.

Studies have been made of the circadian rhythms of a glucorticoid hormone, corticosterone, in the adrenals and blood serum in female Wistar rats from two substrains selected for high (ESTH) and low (ESTL) ability to develop permanent oestrus under constant illumination. Significant changes in parameters of the circadian rhythm of the hormone were observed in animals of the 26th generation of selection. Total alleviation of corticosterone rhythm in the blood was on observed in ESTL rats, while in ESTH animals maximum level of the hormone in the blood was shifted to the dark time. Comparison of a high corticosterone content of the adrenals in ESTL rats with a low concentration in the blood plasma indicates the increase in metabolic clearance of the hormone in animals from this strain. It is suggested that the decreased corticosteroid production in the adrenals of ESTH rats facilitates the development of permanent oestrus under constant illumination.

The functional state of adrenal cortex in patients with chronic relapsing pancreatitis.

The functional state of the adrenal cortex (cortisol and 11-OCS in blood, 17-OCS in 24-hours' urine) was studied in 105 patients with chronic relapsing pancreatitis in the acute state. An increase of adrenal cortex glucocorticoid activity was determined in 46.7% of the patients. A decrease of glucocorticoid activity was observed in 10% of the patients. During remission a tendency towards hormone normalization was noted in most of the patients, except for 5 patients with marked exocrine pancreatic insufficiency in whom the hormone content of adrenal cortex remained low. The type of pancreatitis as well as the severity and duration of the disease were noted to exert no regular impact upon the changes of the indices. Since exacerbation of chronic pancreatitis is frequently accompanied by elevated glucocorticoid activity of the adrenal cortex, the prescription of glucocorticoid hormones in such patients seems undesirable. The administration of such hormones is possible only for the patients with exocrine pancreas insufficiency.

[Adrenal cortex functional activity in pantothenate deficiency and the administration of the vitamin or its derivatives]. / Funktsional'naia aktivnost' kory nadpochechnikov pri nedostatochnosti pantotenata i vvedenii vitamina ili ego proizvodnykh.

Study of the corticosteroid content in the adrenals and blood of rats under pantothenate deficiency has demonstrated a decrease in adrenocortical function. A single administration of pantothenate in a dose of 3.3 mg/kg reduced the influence of hypovitaminosis on the adrenals. The pantothenate derivatives (pantethine, 4'-phosphopantothenate and CoA in particular) injected to intact animals in a single dose equimolar to 3.3 mg/kg calcium pantothenate per kg bw had a marked steroidogenous effect.

[Study of protein synthesis regulating hormones in human milk]. / Issledovanie v zhenskom grudnom moloke gormonov, vliiaiushchikh na sintez belka.

A study was made of the content in human milk of hormones which regulate together with their specific function processes of protein assimilation and synthesis. These hormones (thyroxine, triiodothyronine, 17 beta-estradiol, prolactin, 11-hydroxycorticosteroids) were determined in the first days of lactation, in the most important period in the nutrition of a newborn and in the formation of lactation. Chromatography, radioimmunoassay and fluorometry were used. The aim of the study was to determine the levels of exogenous hormones which influence the metabolism of proteins received by a newborn. During the first 10 days of lactation the content of prolactin in human milk was 154.5-546.0 muunits/ml, that of 17 beta-estradiol 34.2-124.4 pg/ml, that of thyroxine 0.5-1.1 micrograms%, that of triiodothyronine 8.6-20.8 ng%. A possible involvement of milk hormones in the regulation of anabolic processes of a newborn as well as changes in the first 3-6 days of lactation of the prolactin and 17 beta-estradiol content in milk in connection with the regulation of the start of lactation are discussed.

[Effect of alcoholic intoxication in female rats on the functioning of the hypothalamo-hypophyseal-adrenal cortex system of their progeny]. / Vliianie alkogol'noi intoksikatsii krys-samok na sostoianie gipotalamo-gipofizarno-adrenokortikal'noi sistemy ikh potomstva.

Administration of alcohol (20% ethanol) to female rats for the whole pregnancy leads to activation in the progeny of the hypothalamus-pituitary-adrenocortical system (GPACS). This manifests in an increase in corticoliberin production by neurosecretory cells of the hypothalamus, secretion of ACTH by the pituitary, and high production of corticosteroids by the adrenals. Activation of the GPACS which is demonstrable in fetuses and in young rats aged 1-3 days and 1 and 3 months is replaced by this system inhibition by the age of 6 months. Therefore, alcoholic intoxication of the mother's body has a toxic effect on all the components of the GPACS (production of corticoliberin and ACTH, corticosteroid function of the adrenals).

[Genetic problems of stress. V. The genetics of adrenal cortical reactivity in emotional stress in rats]. / Problemy genetiki stressa. Soobshchenie V. Genetika reaktivnosti kory nadpochechnikov pri émotsional'nom stresse u krys.

Genetic analysis of adrenocortical reactivity (the level of plasma corticosterone and secretion of the adrenal gland in vitro) was carried out under conditions of emotional stress in three inbred strains of rats (Wistar, August, Sprague-Dawley) and in their F1 reciprocal hybrids. The basal adrenocortical function was similar in genotypes studied. However, significant genotype-dependent variability was demonstrated under conditions of stress stimulation. The variability of adrenal secretion in vitro was mediated by genes displaying an additive action, while variability in the level of plasma corticosterone included components both of general combining ability and of specific combining ability. Also, the role of a reciprocal effect was noted. The peculiarities of genetic determination of adrenocortical reactivity to emotional stress are discussed.

[Rat adrenal corticosteroid concentration during hypokinesia combined with measured physical exertion]. / Soderzhanie kortikosteroidov v nadpochechnikakh krys v usloviiakh gipokinezii, sochetannoi s dozirovannymi fizicheskimi nagruzkami.

The paper presents data on the content of corticosteroids in adrenals of rats exposed to 46, 53- and 60-day hypokinesia as well as to 60-day hypokinesia combined with graded exercises. It was found that the content of corticosteroids in adrenals remained essentially unaltered after 46-day hypokinesia, decreased after 53-day hypokinesia and increased after 60-day hypokinesia as compared to the appropriate controls. The use of exercises demonstrated good reserve capabilities of the hypophyseal-adrenal system of rats after 60-day hypokinesia and revealed certain changes in regulatory mechanisms of the hypophysis--adrenal cortex system.