Injection Of Hydrocortisone Research Articles

Application of a fluorometric assay to detect caspase activity in thymus tissue undergoing apoptosis in vivo

To date, in vivo apoptosis within the thymus has been assessed using morphological criteria and/or detection of a DNA ladder indicative of oligonucleosomal fragmentation of the DNA. Here, we have used a fluorometric method to investigate activation of the caspase protease family in the thymus following in vivo induction of apoptosis by injection of the synthetic glucocorticoid hydrocortisone. Cleavage of DEVD-MCA by caspase-3 and other group II caspases releases free MCA which can be detected fluorimetrically. We demonstrate a time-dependent increase in DEVD-MCA cleavage activity within this tissue indicating the activation of caspase-3 like enzymes. This activity was inhibited by the specific group II caspase inhibitor DEVD-CHO. The interpretation of increased caspase activity was confirmed by immunoblot analysis to reveal cleavage of the caspase-3 substrate, fodrin. In addition, agarose gel electrophoresis of the DNA yielded a ladder pattern, confirming the occurrence of apoptosis. This study demonstrates that DEVD-MCA cleavage activity may be a useful quantitative method for the analysis of apoptosis in thymus tissue. It is a relatively rapid procedure not requiring thymocyte isolation or gel electrophoresis and detects fairly early biochemical changes occurring during apoptosis. In the present study we have used this method to demonstrate the involvement of caspases in thymocyte apoptotic death induced in vivo by glucocorticoids. Thus, measurement of caspase activity in thymus tissue may have applications for studying the in vivo effects of immunotoxicants.

Complications of Lumbar Puncture with Injection of Hydrosoluble Material

We report two cases of severe disorders after spinal puncture with injection of hydrosoluble material. The first case concerned a 36-year-old woman with intrathecal injection of 125 ml of hydrocortisone acetate. An intracranial occipital hematoma developed. The second case concerned a 26-year-old man with intrathecal injection of contrast media and hydrocortisone. A chemical meningitis occurred. In both cases the natural course was favorable. Both complications are well known but rare. A review of the literature is made with description of the mechanisms. Preventive therapeutic measures are reported.

Increase in Plasma Interleukin-10 Following Hydrocortisone Injection

Increase in Plasma Interleukin-10 Following Hydrocortisone Injection

Regulation of histidase gene expression by glucagon, hydrocortisone and protein-free/high carbohydrate diet in the rat

The effect of glucagon and hydrocortisone was investigated to understand the mechanism of induction of hepatic histidase gene. In this study, glucagon (0.6 mg 100 g body wt d ) was injected to rats fed 10% casein diet. After 3h of the last injection, histidase activity and amount of enzyme were induced by 3 fold and histidase mRNA concentration by 6 fold. Injection of hydrocortisone (2 mg 100g body wt d ) increased 100% histidase activity and mRNA concentration and by 150% the amount of enzyme after 3 h of the last injection. These results indicate that glucagon is a better inductor of histidase gene expression than hydrocortisone. Another purpose of the study was to evaluate if a protein-free/high carbohydrate diet could reverse the induction of Hal expression produced by a high protein diet. Hal activity, amount of enzyme and mRNA concentration was repressed by 68, 88 and 95% respectively by a protein-free/high carbohydrate diet. Injection of glucagon reversed partially the effect of a high carbohydrate diet, however, injection of hydrocortisone under the concentration used in these experiments did not reverse the effect of a high carbohydrate diet. These results support the evidence that hepatic histidase gene expression is probably regulated transcriptionally by hormones.

Dietary Protein Level Regulates Expression of the Mitochondrial Branched-Chain Aminotransferase in Rats

The first step in the degradation of branched-chain amino acids (BCAA) is transamination catalyzed by the branched-chain aminotransferase (BCAT), which is located in extrahepatic tissues. Studies of the effect of dietary protein on BCAT activity have given contradictory results. Therefore, we established the levels of BCAT activity and mitochondrial BCAT (BCATm) mRNA expression in different organs and tissues of rats. We then determined the effect of different levels of dietary protein in well-nourished rats, the effect of feeding a 0.5% casein diet for 5 wk (protein-malnourished rats) and nutritional rehabilitation of these rats with different levels of dietary protein on BCAT activity and BCATm mRNA expression in selected tissues. Finally, the response of tissue BCAT activity and BCATm mRNA levels in rats fed a 10% casein diet and injected with glucagon (4 d) or hydrocortisone (7 d) was determined. The highest concentration of BCATm mRNA was found in stomach, followed by kidney, heart, muscle, brain, skin and lung. Low levels were found in intestine, and no BCATm mRNA was detectable in liver. Although BCAT activity was significantly higher in muscle, kidney and brain from rats adapted to consume a 50% casein diet for 7 h/d for 10 d than in rats fed 6, 18 or 35% casein diets, only muscle had significantly higher levels of BCATm mRNA. In protein-malnourished rats, BCAT activity and BCATm mRNA expression in kidney, muscle and heart were not different from those of rats with free access to an 18% casein diet. Nutritional rehabilitation of the protein-malnourished rats with 50% casein for 21 d significantly increased the BCAT activity and BCATm mRNA expression in muscle. Neither hydrocortisone nor glucagon injection affected BCAT activity or BCATm mRNA concentrations in rat kidney, muscle or heart. We conclude that the nutritional regulation of BCATm is extrahepatic, tissue specific and may involve transcriptional and post-translational mechanisms.

Urinary Excretion of Oestrone Sulphate and Cortisol in Early Pregnant Gilts Treated with Glucocorticoids

The aim of this study was to determine rates of urinary excretion of oestrone sulphate and cortisol in early pregnant gilts that were untreated or treated with either dexamethasone, corn oil or hydrocortisone. Twenty Polish Landrace gilts were used. They were grouped immediately after mating as follows: Experiment I--Group 1 (5 gilts), control animals and Group 2 (5 gilts), injected i.m. with dexamethasone (30 micrograms/kg) at 12-h intervals from day 13 to day 22 of pregnancy; Experiment II--Group 3 (5 gilts), injected i.m. with corn oil at 12-h intervals from day 13 to day 22 of pregnancy and Group 4 (5 gilts), injected i.m. with hydrocortisone acetate (250 mg) at 12-h intervals from day 11 to day 20 of pregnancy. Gilts were placed in metabolic cages, and 24-h urine aliquots were collected from day 6 to day 32 of pregnancy. On days 34-36 of pregnancy gilts were slaughtered and clinical data were collected. Rates of urinary excretion of oestrone sulphate and cortisol were determined by enhanced chemiluminescence immunoassays. The urinary excretion of oestrone sulphate expressed in nmol/24 h and mumol/mol creatinine were significantly correlated. There was no correlation between cortisol expressed in nmol/24 h and mumol/mol creatinine (p > 0.5). A first significant increase of urinary oestrone sulphate excretion, expressed in nmol/24 h, on days 13-14 and a second one on days 19-20 of gestation occurred in control untreated and oil-treated gilts. The urinary excretion of oestrone sulphate reached maximum values between days 25 and 32 of gestation. In dexamethasone-treated gilts cortisol excretion significantly decreased on day 16, i.e. 3 days after injections of dexamethasone had commenced. The treatment with hydrocortisone resulted in a significantly increased cortisol excretion after the last injection of hydrocortisone. There were no relations between levels of urinary oestrone sulphate excretion expressed in nmol/24 h and the number of foetuses. When the urinary excretion of oestrone sulphate was expressed in mol/mol creatinine we found a positive relation between concentrations on day 20 of pregnancy and the number of foetuses. In one untreated gilt with a relatively high urinary excretion of cortisol (more than 200 nmol/24 h) a lower number of foetuses was found at autopsy. In conclusion, both dexamethasone and hydrocortisone treatment seemed to delay the first observed peak in oestrone sulphate in gilts without affecting the embryonic survival and the number of viable foetuses.

Effect of testosterone and cortisol administration on the reproductive tract of male Antechinus stuartii (Marsupialia).

The life history of Antechinus stuartii, a marsupial, is highly synchronized and culminates in a brief mating period that is followed by complete male mortality. The accessory reproductive tracts of male A. stuartii enlarge in association with testosterone and cortisol hormone concentrations, but this appears to be unrelated to the spermatogenic cycle. The present study examined the effects of testosterone and cortisol on the male reproductive tract. Four groups of adult males from May (when plasma testosterone and cortisol concentrations are low) were given depot injections of testosterone esters or synthetic cortisol in doses that mimic concentrations found in males in the breeding period (August). Males were given either saline, testosterone only, cortisol only, or testosterone plus cortisol. Experimental groups did not differ in the seminiferous tubule morphology. However, the cells from the caudal end of the epididymides of both testosterone groups were considerably hypertrophied compared with males treated with saline or cortisol only. Testosterone treatment significantly increased prostate and bulbourethral gland mass, although addition of cortisol to the testosterone administration diminished this effect. The morphology of the accessory reproductive tract of males treated with either saline or cortisol only was similar to that of untreated males at the same time of year, and the morphology of the accessory reproductive tract of males treated with testosterone plus cortisol was similar to that of untreated males in the breeding season. Like some other marsupials, the spermatogenic cycle in A. stuartii is apparently not correlated with androgen activity, while the accessory reproductive tract is affected by androgens.

Success of glucocorticoid replacement therapy on fertility in two adult males with 21-CAH homozygote classic form

A normal gonadal maturation with normal fertility are some of the major goals of long-term replacement therapy in adult males with Congenital Adrenal Hyperplasia (CAH). We describe here two young men, G.O. (case A, 23 years old) and S.S.(case B, 24 years old), both with a well defined diagnosis of CAH due to 21-hydroxylase deficiency classic homozygote form (21-CAH). In case A the diagnosis of the 21-CAH classic virilizing form was made at 3 years of age. The patient has undergone glucocorticoid therapy and is now 170 cm tall; all his hormonal findings are within the normal range. The semen analysis has shown a good fertility potential, with a slight modification when the patient decided to discontinue the therapy. In case B the diagnosis of the 21-CAH salt wasting form was performed at 9 days of age. The patient was initially treated with i.v. normal saline solution and a daily i.m. injection of hydrocortisone and, subsequently, with mineral and glucocorticoid replacement therapy po. A satisfactory adult stature (165 cm) was attained. The patient is still on therapy, with a good hormonal profile. The semen analysis has shown an apparently normal fertility. In conclusion, our experience in adult males with 21-CAH, who have been administered prompt and adequate replacement therapy, shows that these patients can attain normal quality of life, satisfactory growth and development, normal sexual maturation and activity, and adequate sperm fertilizing ability, thereby supporting the usefulness of continuing this therapy during adult age.

Thymic aging in ICR female mice is suspended by prolonged hydrocortisone exposure.

Age-related decline of the thymus in ICR female mice was studied following long-term (three month) weekly exposure to hydrocortisone acetate. When examined one week after cortisone injections, the well-known thymic atrophy was observed. Five weeks after 12 hydrocortisone injections, the cortical volume fraction (Vc), cortical/medullary ratio (C/M), the number of thymocytes and CD4/CD8 profiles were in the range that characterizes younger mice, compared with PBS-injected mice, uninjected controls, or mice given a single hydrocortisone injection 5 weeks earlier. It seems as if thymic involution with age was suspended during the period of glucocorticoid exposure.

Mast cells/eosinophilic granule cells of salmonids: staining properties and responses to noxious agents

Observations were made on mast cells/eosinophilic granule cells in swimbladder tissue spreads and sections of gills and intestinal tissues from species of the generaSalmoOncorhynchusSalvelinusCoregonusThymallus. Some individuals had been reared in captivity and others were caught in rivers or lakes, and both apparently healthy fish and fish with persistent inflammation, due to helminths or unknown causative agents, were included in the study. Acute responses to noxious agents were studied in swimbladder tissue spreads after intraperitoneal injections of inactivatedAeromonas salmonicida, compound 48/80 and hydrocortisone. The tissue spreads were fixed in ethanol and stained with thionin. Other tissues were fixed in a solution containing 4% formaldehyde and 5% acetic acid in methanol, and stained with May-Grünwald Giemsa combination dye, haematoxylin and eosin, or Alcian blue. Intestinal tissue histamine was assayed fluorometrically, and vascular responses to histamine and compound 48/80 were studied in perfused gill preparations. The staining properties of salmonid mast cells/eosinophilic granule cells resembled those of mammalian mucosal mast cells and globule leucocytes, with both acidophilic and basophilic components in their granules. May-Grünwald Giemsa staining revealed that in cells found in connective tissues the basophilic character was dominant, whereas the acidophilic character was most marked in those present in epithelia. The granule cells in swimbladder tissue spreads stained metachromatically with alcoholic thionin. Intraperitoneal injections of inactivatedA. salmonicidaproduced acute inflammatory reactions, with degranulation of mast cells/eosinophilic granule cells, in tissues of the swimbladder. Degranulation of the granule cells was also noticed after injection of compound 48/80. Massive degranulation of mast cells/eosinophilic granule cells in the swimbladder wall, followed by an acute inflammatory reaction, was induced by intraperitoneal injections of hydrocortisone. Persistent inflammation, e.g. in tissues infected with helminths, was accompanied by recruitment of mast cells/eosinophilic granule cells. Presence of many or few mast cells/eosinophilic granule cells in tissues of the intestine seemed to have no influence on the content of histamine, which was always low. Compound 48/80 produced increased resistance in the perfused branchial vascular bed, but effects of histamine were slight or completely absent. The responses of mast cells/eosinophilic granule cells of salmonids in acute and persistent inflammation, as revealed in the present investigation, are similar to the known responses of mammalian mast cells. Since their staining properties are also similar, the term ‘mast cell’ should be adequate.

Cortisol Enhances non-REM Sleep and Growth Hormone Secretion in Elderly Subjects

Aging is accompanied by a continuous decline in slow wave sleep (SWS) and in growth hormone (GH) secretion, particularly during the sleeping period. Because short-term pulsatile administration of cortisol increases GH release and SWS in young adults, we wondered whether similar effects can be induced also in elderly men. Hourly injections of cortisol between 1700 and 600 h increased stage 2 and SWS and decreased rapid eye movement sleep. Spectral analysis revealed significant increases in delta and theta power. Cortisol infusions increased the GH secretion prior to sleep onset, but remained largely unchanged during sleep. Thus, sleep EEG and GH release are modulated by cortisol administration in a manner similar to that in young subjects, but to a lesser extent. The stimulatory effect of cortisol on both GH release and SWS points to a mechanism involving glucocorticoid-enhanced production and release of GH-releasing hormone that activates pituitary GH release and simultaneously antagonizes the effects of corticotropin-releasing hormone and somatostatin.

COMPLEX RELATIONS BETWEEN CORTICOSTEROIDS AND THE ADHESIVE STATE OF THE WHITE BLOOD CELLS IN THE PERIPHERAL BLOOD: FURTHER ANALYSIS OF THE STATE OF LEUKOCYTE/ADHESIVENESS AGGREGATION IN THE PERIPHERAL BLOOD AS A MARKER OF STRESS

Stress is associated with an increment in the concentration of cortisol, a leukocytotic response and increased state of leukocyte adhesiveness/aggregation (LAA) in the peripheral blood. We investigated the interrelationship between these three parameters in individuals under various conditions of stress. These included 123 emergency room patients with chest pain who were also classified in regard to the presence or absence of heart failure; 15 men in the fertility clinic donating semen and 16 women in the operating room before induction of anaesthesia for dilatation and curettage. A low (r=0.35) but significant (p<0.001) correlation was noted between cortisol level, leukocyte count, and per cent of aggregated leukocytes in the peripheral blood in the first group (ischaemic heart disease) but not the fertility clinic or operating room patients. A complementary study in dogs which received an intravenous hydrocortisone injection revealed neither an increased leukocyte number in the peripheral blood nor an increment in the adhesive state. In vitro studies confirmed the depressant effect of high concentrations of hydrocortisone on cell adhesiveness despite an increment in the expression of various adhesive proteins on the surface of the peripheral blood leukocytes. We concluded that the increased state of leukocyte adhesiveness noted by us in the past in patients under stress is not necessarily related to the presence of an increased cortisol concentration, and that factors other than corticosteroid level should be explored as possible triggers of the LAA response during stress. © 1997 by John Wiley & Sons, Ltd.

COMPLEX RELATIONS BETWEEN CORTICOSTEROIDS AND THE ADHESIVE STATE OF THE WHITE BLOOD CELLS IN THE PERIPHERAL BLOOD: FURTHER ANALYSIS OF THE STATE OF LEUKOCYTE/ADHESIVENESS AGGREGATION IN THE PERIPHERAL BLOOD AS A MARKER OF STRESS

Stress is associated with an increment in the concentration of cortisol, a leukocytotic response and increased state of leukocyte adhesiveness/aggregation (LAA) in the peripheral blood. We investigated the interrelationship between these three parameters in individuals under various conditions of stress. These included 123 emergency room patients with chest pain who were also classified in regard to the presence or absence of heart failure; 15 men in the fertility clinic donating semen and 16 women in the operating room before induction of anaesthesia for dilatation and curettage. A low (r=0.35) but significant (p<0.001) correlation was noted between cortisol level, leukocyte count, and per cent of aggregated leukocytes in the peripheral blood in the first group (ischaemic heart disease) but not the fertility clinic or operating room patients. A complementary study in dogs which received an intravenous hydrocortisone injection revealed neither an increased leukocyte number in the peripheral blood nor an increment in the adhesive state. In vitro studies confirmed the depressant effect of high concentrations of hydrocortisone on cell adhesiveness despite an increment in the expression of various adhesive proteins on the surface of the peripheral blood leukocytes. We concluded that the increased state of leukocyte adhesiveness noted by us in the past in patients under stress is not necessarily related to the presence of an increased cortisol concentration, and that factors other than corticosteroid level should be explored as possible triggers of the LAA response during stress. © 1997 by John Wiley & Sons, Ltd.

In vitro measurement of orthodontic tooth movement in rats given beta-aminopropionitrile or hydrocortisone using a time-lapse videotape recorder.

In vitro tooth movement of rat molars in response to an orthodontic force was recorded using a time-lapse videotape recorder and analysed by a computer system. Rats received daily s.c. injections of beta-aminopropionitrile (BAPN, 300 mg/kg/day) or hydrocortisone (10 mg/kg/day) for a period of 7 days. After drug administration, the animals were killed and the mandibles dissected. The jaw was then held under a stereomicroscope with a haemostatic clamp and an elastic band was inserted between the first and second molars. The movements of reference points on the occlusal surfaces of the first and second molars were recorded for 20 hours using a time-lapse videotape recorder. Mesiolingual movement of the first molar and distobuccal movement of the second molar were observed. During the experimental period, the greatest amount of total tooth movement in the first and second molars was seen in the group pretreated with BAPN, less movement was observed in the control group, and the group pretreated with hydrocortisone exhibited the least amount of movement. The highest rates of tooth movement were observed during the initial hour in each of the groups, and decreased thereafter. The initial rates of movement were also greatest in the BAPN group, less in the control group, and least in the hydrocortisone group. These results indicate that treatment with BAPN accelerated experimental tooth movements in vitro and hydrocortisone treatment inhibited the movements, suggesting that, although a part of the tooth movement measured in this experiment was due to deformation of the alveolar bone, the mechanical properties of the periodontal ligament play an important role in the regulation of orthodontic tooth movement.

Effects of prenatal stress on suckling calves.

Pregnant Brahman cows (n = 42), bred to either Brahman or Tuli bulls, were randomly assigned to one of three treatments: 1) transported in a stock trailer for 24.2 km, unloaded at a second farm and penned for 1 h, and then returned to the original farm (TRANS); 2) i.v. injection of ACTH, 1 IU/kg BW (ACTH); or 3) walked through the handling facilities (SHAM). Treatments were initiated on d 60 and repeated at 80, 100, 120, and 140 d of gestation. The calves from these cows were subjected to tests to measure their capacity to react to stress. In Test 1, Tuli-sired calves were restrained at 10 and 150 d of age for 3.5 h. In Test 2, Brahman-sired calves were restrained for 3.5 h and given an injection of ACTH (.125 IU ACTH/kg of BW). In Test 3, Test-2 calves were restrained at 180 d of age and hot-iron branded. In Test 4, Test-1 calves were restrained at 180 d of age and given an injection of cortisol (6.7 ng/kg BW) to estimate cortisol clearance rate. During all tests, calves were restrained for 3.5 h, and heart rates were recorded and blood samples were taken at -15, 0, 15, 30, 45, 60, 90, 120, and 180 min. The 10- and 150-d-old TRANS calves maintained greater plasma cortisol in Test 1 (restraint) than the ACTH and SHAM calves (P < .01). The ACTH challenge (Test 2) increased plasma cortisol and ACTH, but cow treatment did not alter the response (P > .4). In response to branding (Test 3), the TRANS, ACTH, and SHAM calves' overall mean plasma cortisol was not affected by treatment (52, 51, and 43 +/- 3 ng/mL, respectively; P > .1), nor was the calves' overall heart rate (91, 94, and 86 +/- 3 beats/min, respectively; P > . 1). In Test 4, TRANS calves cleared plasma of cortisol at a slower rate than did the SHAM calves (P < .01), but not the ACTH calves (261, 374, and 473 +/- 50 mL/min, respectively; P > .1). The TRANS calves had an overall greater heart rate than did the ACTH or the SHAM calves (91, 79, and 77 +/- 2 beats/min, respectively; P < .001). Exposing cows to repeated transportation stress during gestation altered their calf's physiological response to stress, and these alterations could have a profound influence on the calfs ability to adapt to stress, thereby influencing its welfare. Further research should examine the growth, immune function, and reproductive function of prenatally stressed calves to determine whether these changes in plasma cortisol are beneficial or deleterious.

The effect of intraarticular hydrocortisone injection on the articular cartilage of rabbits.

We investigated the effect of hydrocortisone on the articular cartilage of the knee in rabbits. 27 New Zealand white rabbits were injected intraarticularly with 25, 50 or 100 mg betamethasone acetate in 2 or 4 weekly intervals. Control animals were injected with normal saline and demonstrated no histological changes in the articular cartilage. Hydrocortisone administration was associated with increased cell size, as well as an increased stain density in the cytoplasm surrounding vacuoles. In addition, loss of cell organelles was also observed. High dose of hydrocortisone was associated with an obvious loss of cell shape and distortion of the cell membrane and nucleus. The magnitude of histological changes, found under light and electron microscopy, were proportional to the amount of hydrocortisone injected. Our findings strongly indicate that intraarticular injection of hydrocortisone alters the shape of articular cartilage chondrocytes, producing abnormal changes in the cytoplasm and nucleus and leading to cell degeneration.

То the problem of temporary disablement examination in the treatment of bursitises, hydromas and ganglions

As many as 97 cases of ulnar bursitises, 24 prepatellar and 13 popliteal hygromas along with 22 cases of ganglions on arms and legs, the efficiency of their treatment methods depending on temporary disablement period are analyzed. The great efficiency of the ultrasound treatment with hydrocortisone, liquid removal using punctures, the injection of hydrocortisone with Novocain to the synovial bursa cavity and immobilization of full value is revealed.

Prenatal Glucocorticoid Therapy Reverses Pulmonary Immaturity in Congenital Diaphragmatic Hernia in Fetal Sheep

To assess the feasibility of conducting clinical trials of prenatal steroid therapy for congenital diaphragmatic hernia (CDH) in humans, the authors tested whether prenatal glucocorticoid, currently the standard treatment to minimize respiratory distress syndrome in premature infants, might improve the pulmonary immaturity in severe CDH in a large animal model. The authors have used the nitrofen-induced rat model of CDH, which demonstrates immature lungs by biochemical, morphometric, and molecular biologic criteria. They also have shown that the lethally immature lungs of the full-term CDH rats can be improved by biochemical, morphometric, physiologic, and molecular criteria by treating the mothers with parenteral steroids at doses extrapolated from the current therapy used to accelerate lung development of premature human babies. During a 3-year period in 88 fetal sheep, 1) left-sided diaphragmatic hernias were created surgically at varying gestational ages (day 78-90; term = 142-145 days) and size to maximize severity (n = 45), 2) placement and design of indwelling fetal intravenous catheters were optimized (n = 13), and 3) timing and dosage of cortisol administration were determined (n = 17). As a result, diaphragmatic hernias were created on day 80, intravenous catheters were placed on day 120, and twice-daily intravenous cortisol injections (n = 8) or saline as the control (n = 5) were administered (days 133-135). Lambs were delivered on day 136 via cesarean section to avoid steroid-induced abortion; vascular access was obtained, and the fetuses were ventilated at standard settings. Physiologic data were collected, and lungs were harvested for biochemical and histologic analysis. Significant improvements were measured in postductal arterial oxygen pressure ([PaO2] 38 +/- 6 mmHg after cortisol therapy compared with 20 +/- 3 mmHg for saline controls; p = 0.002) and in dynamic compliance (0.42 +/- 0.05 mL/cm H2O vs. 0.29 +/- 0.01 mL/cm H2O; p = 0.01). Lung glycogen levels in the right lung of the cortisol group were significantly better than controls (4.6 +/- 0.3 mg/g lung vs. 6.8 +/- 0.4 mg/g; p = 0.002), as were protein/DNA levels (8.3 +/- 0.9 mg/mg vs. 14.5 +/- mg/mg; p < 0.05). Striking morphologic maturation of airway architecture was observed in the treated lungs. Prenatal glucocorticoids correct the pulmonary immaturity of fetal sheep with CDH by physiologic, biochemical, and histologic criteria. These data, combined with previous small animal studies, have prompted the authors to initiate a prospective phase I/II clinical trial to examine the efficacy of prenatal glucocorticoids to improve the maturation of hypoplastic lungs associated with CDH.

Evidence That Cortisol May Protect against the Immediate Effects of Stress on Circulating Leukocytes in the Trout

Rainbow trout stressed by an intraperitonal injection of saline displayed reduced phagocytic activity of their spleen and head-kidney macrophages within 3 hr. Phagocytic activity was similarly depressed by injecting noradrenalin, but was maintained in fish injected with the adrenergic blocking agent phentolamine, suggesting that endogenous catecholamines are involved in this stress response. Since stress may increase the number of circulating granulocytes, it is proposed that noradrenalin, released during stress, causes the liberation of active macrophages from the lymphocytic tissue, the remaining macrophages therefore showing a lowered phagocytic index. Cortisol injection, like phentolamine,preventedthe depressive effect of stress on the phagocytic index but did not antagonize the depressive effect of exogenous noradrenalin. It is suggested that the stress-induced release of endogenous catecholamines may be prevented by cortisol. Injection stress caused a decline in the number of circulating lymphocytes/thrombocytes, indicating their retrafficking into some other tissue. This was opposed by cortisol and by high doses of noradrenalin. It is proposed that cortisol or noradrenalin may oppose, directly or indirectly, the expression of adhesion molecules which are normally induced after stress.

Differential effects of angiostatic steroids and dexamethasone on angiogenesis and cytokine levels in rat sponge implants.

1. Subcutaneous implantation of sterile polyether sponges elicited a reproducible neovascular response in rats, as determined by blood flow measurement with a 133Xe clearance technique and confirmed histologically. This model was used to monitor the levels of two cytokines during angiogenesis and to compare the activities of angiostatic steroids and anti-inflammatory steroids. 2. Initial experiments followed the neovascular development over a 20-day period. Daily local injection of hydrocortisone caused a dose-dependent (0.5, 5 and 50 micrograms per sponge) inhibition of the basal sponge-induced angiogenesis. However, daily systemic treatment of hydrocortisone (2, 10 and 50 mg kg-1, s.c.) was less effective at inhibiting angiogenesis, and this inhibition was not sustained by day 20 after sponge implantation. 3. To investigate the involvement of cytokines during the course of angiogenesis, we measured the endogenous levels of tumour necrosis factor-alpha (TNF-alpha) and interleukin 6 (IL-6) in sponge implants. Levels of IL-6 and TNF-alpha peaked at day 7 and day 11 after implantation, respectively. These cytokine levels subsided through the completion of angiogenesis by day 20. 4. Subsequent experiments were carried out over a 14-day period. Among the three angiostatic steroids tested, U-24067 (6 alpha-fluoro-17,21 - dihydroxy-16 alpha-methylpregna -4,9(11)-diene-3,20-dione-21-acetate) showed a dose-dependent inhibition (0.5, 5 and 50 micrograms per sponge per day) of sponge-induced angiogenesis. Tetrahydro-S was also effective at 5 micrograms doses, but medroxyprogesterone failed to affect the angiogenic response. None of these steroids caused atrophies of the spleen and thymus. 5. Daily local injection of dexamethasone (0.5 microgram per sponge) inhibited the basal sponge-induced angiogenesis almost completely. Although higher doses of dexamethasone (5 and 50 micrograms per sponge) did not produce further inhibition of angiogenesis, they caused severe spleen and thymus weight losses, indicative of immunosuppression. 6. At the daily dose of 5 micrograms per sponge, dexamethasone inhibited angiogenesis and produced a marked reduction in the levels of TNF-alpha and IL-6 at day 14. In contrast, hydrocortisone, U-24067 and tetrahydro-S did not influence the levels of TNF-alpha and IL-6. 7. We concluded that the anti-angiogenic activity of angiostatic steroids and anti-inflammatory steroids in the rat sponge model is independent of their ability to reduce the production of TNF-alpha and IL-6. The differential effects of angiostatic and anti-inflammatory steroids suggest that U-24067 and its derivatives may have therapeutic potential in the management of angiogenic diseases such as rheumatoid arthritis.