Increased Corticosteroid Levels Research Articles

Typical gene expression profile of pseudorabies virus reactivation from latency in swine trigeminal ganglion.

Pseudorabies virus (PRV) establishes a lifelong latent infection in swine trigeminal ganglion (TG) following acute infection. Increased corticosteroid levels, due to stress, increases the incidence of reactivation from latency. Muscle injection combined with intravenous deliver of the synthetic corticosteroid dexamethasone (DEX) consistently induces reactivation from latency in pigs. In this study, PRV-free piglets were infected with PRV. Viral shedding in nasal and ocular swabs demonstrated that PRV infection entered the latent period. The anti-PRV antibody was detected by enzyme-linked immunosorbent assay and the serum neutralization test, which suggested that the PRV could establish latent infection in the presence of humoral immunity. Immunohistochemistry and viral genome detection of TG neurons suggested that PRV was reactivated from latency. Viral gene expressions of IE180, EP0, VP16, and LLT-intron were readily detected at 3-h post-DEX treatment, but gB, a γ1 gene, was not detectable. The differentially expressed phosphorylated proteins of TG neurons were analyzed by ITRAQ coupled with LC-MS/MS, and p-EIF2S2 differentially expression was confirmed by western blot assay. Taken together, our study provides the evidence that typical gene expression in PRV reactivation from latency in TG is disordered compared with known lytic infection in epithelial cells.

Two Pioneer Transcription Factors, Krüppel-Like Transcription Factor 4 and Glucocorticoid Receptor, Cooperatively Transactivate the Bovine Herpesvirus 1 ICP0 Early Promoter and Stimulate Productive Infection.

An important site for bovine herpesvirus 1 (BoHV-1) latency is sensory neurons within trigeminal ganglia (TG). The synthetic corticosteroid dexamethasone consistently induces BoHV-1 reactivation from latency. Expression of four Krüppel-like transcription factors (KLF), i.e., KLF4, KLF6, PLZF (promyelocytic leukemia zinc finger), and KLF15, are induced in TG neurons early during dexamethasone-induced reactivation. The glucocorticoid receptor (GR) and KLF15 form a feed-forward transcription loop that cooperatively transactivates the BoHV-1 immediate early transcription unit 1 (IEtu1) promoter that drives bovine infected cell protein 0 (bICP0) and bICP4 expression. Since the bICP0 gene also contains a separate early (E) promoter, we tested the hypothesis that GR and KLF family members transactivate the bICP0 E promoter. GR and KLF4, both pioneer transcription factors, cooperated to stimulate bICP0 E promoter activity in a ligand-independent manner in mouse neuroblastoma cells (Neuro-2A). Furthermore, GR and KLF4 stimulated productive infection. Mutating both half GR binding sites did not significantly reduce GR- and KLF4-mediated transactivation of the bICP0 E promoter, suggesting that a novel mechanism exists for transactivation. GR and KLF15 cooperatively stimulated bICP0 activity less efficiently than GR and KL4: however, KLF6, PLZF, and GR had little effect on the bICP0 E promoter. GR, KLF4, and KLF15 occupied bICP0 E promoter sequences in transfected Neuro-2A cells. GR and KLF15, but not KLF4, occupied the bICP0 E promoter at late times during productive infection of bovine cells. Collectively, these studies suggest that cooperative transactivation of the bICP0 E promoter by two pioneer transcription factors (GR and KLF4) correlates with stimulating lytic cycle viral gene expression following stressful stimuli.IMPORTANCE Bovine herpesvirus 1 (BoHV-1), an important bovine pathogen, establishes lifelong latency in sensory neurons. Reactivation from latency is consistently induced by the synthetic corticosteroid dexamethasone. We predict that increased corticosteroid levels activate the glucocorticoid receptor (GR). Consequently, viral gene expression is stimulated by the activated GR. The immediate early transcription unit 1 promoter (IEtu1) drives expression of two viral transcriptional regulatory proteins, bovine infected cell protein 0 (bICP0) and bICP4. Interestingly, a separate early promoter also drives bICP0 expression. Two pioneer transcription factors, GR and Krüppel-like transcription factor 4 (KLF4), cooperatively transactivate the bICP0 early (E) promoter. GR and KLF15 cooperate to stimulate bICP0 E promoter activity but significantly less than GR and KLF4. The bICP0 E promoter contains enhancer-like domains necessary for GR- and KLF4-mediated transactivation that are distinct from those for GR and KLF15. Stress-induced pioneer transcription factors are proposed to activate key viral promoters, including the bICP0 E promoter, during early stages of reactivation from latency.

Bovine Herpesvirus 1 Counteracts Immune Responses and Immune-Surveillance to Enhance Pathogenesis and Virus Transmission.

Infection of cattle by bovine herpesvirus 1 (BoHV-1) can culminate in upper respiratory tract disorders, conjunctivitis, or genital disorders. Infection also consistently leads to transient immune-suppression. BoHV-1 is the number one infectious agent in cattle that is associated with abortions in cattle. BoHV-1, as other α-herpesvirinae subfamily members, establishes latency in sensory neurons. Stressful stimuli, mimicked by the synthetic corticosteroid dexamethasone, consistently induce reactivation from latency in latently infected calves and rabbits. Increased corticosteroid levels due to stress have a two-pronged effect on reactivation from latency by: (1) directly stimulating viral gene expression and replication, and (2) impairing antiviral immune responses, thus enhancing virus spread and transmission. BoHV-1 encodes several proteins, bICP0, bICP27, gG, UL49.5, and VP8, which interfere with key antiviral innate immune responses in the absence of other viral genes. Furthermore, the ability of BoHV-1 to infect lymphocytes and induce apoptosis, in particular CD4+ T cells, has negative impacts on immune responses during acute infection. BoHV-1 induced immune-suppression can initiate the poly-microbial disorder known as bovine respiratory disease complex, which costs the US cattle industry more than one billion dollars annually. Furthermore, interfering with antiviral responses may promote viral spread to ovaries and the developing fetus, thus enhancing reproductive issues associated with BoHV-1 infection of cows or pregnant cows. The focus of this review is to describe the known mechanisms, direct and indirect, by which BoHV-1 interferes with antiviral immune responses during the course of infection.

Bovine Herpesvirus 1 Regulatory Proteins bICP0 and VP16 Are Readily Detected in Trigeminal Ganglionic Neurons Expressing the Glucocorticoid Receptor during the Early Stages of Reactivation from Latency

Bovine herpesvirus 1 (BHV-1) establishes a lifelong latent infection in sensory neurons following acute infection. Increased corticosteroid levels, due to stress, increases the incidence of reactivation from latency. Within minutes, corticosteroids activate the glucocorticoid receptor and transcription of promoters containing a glucocorticoid receptor element. A single intravenous injection of the synthetic corticosteroid dexamethasone consistently induces reactivation from latency in calves. Lytic cycle viral gene expression is detected within 6 h after dexamethasone treatment of calves latently infected with BHV-1. Cellular transcription factors are induced by dexamethasone in trigeminal ganglionic neurons within 1.5 h after dexamethasone treatment, suggesting they promote viral gene expression during the early phases of reactivation from latency, which we operationally defined as the escape from latency. In this study, immunohistochemistry was utilized to examine viral protein expression during the escape from latency. Within 1.5 h after dexamethasone treatment, bICP0 and a late protein (VP16) were consistently detected in a subset of trigeminal ganglionic neurons. Most neurons expressing bICP0 also expressed VP16. Additional studies revealed that neurons expressing the glucocorticoid receptor also expressed bICP0 or VP16 at 1.5 h after dexamethasone treatment. Two other late proteins, glycoprotein C and D, were not detected until 6 h after dexamethasone treatment and were detected in only a few neurons. These studies provide evidence that VP16 and the promiscuous viral trans-activator (bICP0) are expressed during the escape from latency, suggesting they promote the production of infectious virus in a small subset of latently infected neurons.

Stimulatory effect of SOM230 on human and rat adrenal corticosteroid secretion in vitro

SOM230 (pasireotide, Signifor), a recently developed somatostatin analog, has been tested in ACTH-secreting pituitary tumors with promising results. No study has yet evaluated whether this analog also directly affects adrenal steroid production. The aim of the current study was to evaluate whether SOM230 modulates corticosteroid secretion by normal adrenals in vitro. Primary cultures from normal human and rat adrenals were incubated with 10–100nM SOM230 with and without 10nM ACTH. Dose–response studies with 1nM–1μM SOM230 were performed on rat adrenals. Cortisol/corticosterone levels in medium were measured after 4 and 24h. SOM230 (10nM) significantly increased corticosteroid levels after 24h incubation in both human (36.4±0.43ng/well vs 27.7±3.17ng/well, p<0.05) and rat (16.2±1.16ng/well vs 11.6±0.92ng/well p<0.05) adrenals; lesser effects were observed with 100nM SOM (33.4±2.59ng/well vs 27.7±3.17ng/well p<0.05; 13.4±0.82ng/well vs 11.6±0.92ng/well, N.S. vs baseline secretion for human and rat adrenals, respectively). Dose–response curves confirmed maximal effect at 10nM SOM230. The corticosteroid secretory response to ACTH was unaffected by SOM230 co-incubation. In conclusion, SOM230 exerts a moderate stimulatory effect on adrenal corticosteroid secretion in vitro. This argues against a direct adrenal involvement in the clinical efficacy of SOM230 in patients with ACTH-secreting pituitary tumors and widens the known range of action of SOM230.

Molarless condition suppresses proliferation but not differentiation rates into neurons in the rat dentate gyrus

The dentate gyrus (DG) of the hippocampal complex is one of the few areas of the rodent brain where neurogenesis continues throughout adulthood. We investigated the effects of the molarless condition on cell proliferation, rate of differentiation into neurons in the subgranular zone of the DG, and plasma corticosterone levels. The molarless condition decreased cell proliferation in the DG and increased plasma corticosterone levels. Approximately 80% of newly generated cells differentiated into neurons and the remaining 20% of the cells differentiated into astrocytes. These ratios were not significantly different between control and molarless rats. In conclusion, the rates of neurogenesis and gliogenesis in the DG are suppressed by the molarless condition, and this suppression might be associated with the increased corticosteroid levels in molarless subjects.

Declarative Memory after Stress in Humans: Differential Involvement of the β-Adrenergic and Corticosteroid Systems

To determine the role played by the beta-adrenergic and corticosteroid systems in the modulatory effects of stress on declarative memory function, 42 young men were administered a placebo, propranolol (beta-adrenergic blocker), or metyrapone (corticosteroid synthesis inhibitor) before being submitted to a psychological stress protocol. Immediately after stress, subjects viewed a neutral story, unrelated to the stressor. Short- (5 min post learning) and long-term (1 wk post learning) recall of the story was assessed. Placebo and propranolol groups showed significant stress-related increases in corticosteroid levels, whereas metyrapone prevented corticosteroid reactivity to the stressor. Stress triggered significant elevations in cardiac activity (heart rate, systolic and diastolic blood pressure levels) in all three groups, with the metyrapone group showing the strongest elevation in heart rate levels in response to stress. Compared with placebo, propranolol had no effect on short- and long-term recall of the story learned after stress, whereas metyrapone impaired short-term recall of the story, with no further effects on long-term declarative memory. These results suggest that, contrary to the beta-adrenergic system, the corticosteroid system is implicated in declarative memory function after stress in humans.

Effects of Chronic Stress on Structure and Cell Function in Rat Hippocampus and Hypothalamus

It has become increasingly clear that the increase in corticosteroid levels, e.g. after a brief stressor induce molecular and cellular changes in brain, including the hippocampal formation. These effects eventually result in behavioral adaptation. Prolonged exposure to stress, though, may lead to mal-adaptation and even be a risk factor for diseases like major depression in genetically predisposed individuals. We conducted a series of experiments where changes in brain function were examined after 3 weeks of unpredictable stress. After unpredictable stress, inhibitory input to neurons involved in the hypothalamus-pituitary-adrenal (HPA) axis regulation was suppressed, which may dysregulate the axis and lead to overexposure of the brain to glucocorticoids. Furthermore, glutamate transmission in the dentate gyrus (DG) was enhanced, possibly through transcriptional regulation of receptor subunits. Combined with enhanced calcium channel expression this could increase vulnerability to cell death. Neurogenesis and apoptosis in the dentate were diminished. Synaptic plasticity was suppressed both in the dentate and CA1 area. Collectively, these effects may give rise to deficits in memory formation. Finally, we observed reduced responses to serotonin in the CA1 area, which could contribute to the onset of symptoms of depression in predisposed individuals. All of these endpoints provide potential targets for novel treatment strategies of stress-related brain disorders.

Semelparity in a large marsupial.

Complete mortality of males after mating is known in several small dasyurid and didelphid species (up to 300g) and has previously been suggested to be a consequence of their small size and their inability to sequester sufficient fat reserves for an intense rut in the winter. Males of these species use increased corticosteroid levels to allow protein catabolism, enabling them to support their mating effort with other body reserves. However, increased corticosteroid levels have negative consequences such as anaemia, gastrointestinal ulceration, immune suppression and disease. The Australian dasyurid Dasyurus hallucatus shows complete male die off after mating in tropical savannah, yet males of this species may weigh as much as 1120 g and continue to eat during the rut. Die off in D. hallucatus shows many similarities to that in the smaller species including weight loss, fur loss, parasite infestation, increased testosterone levels and anaemia. However, in contrast to smaller species, there is no evidence of elevated corticosteroid levels or gastrointestinal ulceration. Consequently, the phenomenon of male die off after mating lacks a universal explanation.

Cavernous sinus sampling

I NTERVENTIONAL neuroradiology now plays a role in the diagnosis of Cushings disease. The disease is characterized by an increased production of corticosteroids, leading to the classic features: increase in central body weight with sparing of the extremities, increased body hair, pliable skin, hypertension, and hyperglycemia. The release of corticosteroids from the adrenal gland is controlled by a negative feedback mechanism. Corticotropin is released from the hypothalamus and travels to the anterior pituitary gland via a portal venous system. Corticotropin stimulates the release of adrenocorticotrophic hormone (ACTH) from the anterior pituitary gland. This hormone stimulates the production and secretion of corticosteroids from the adrenal cortex. The increase of corticosteroid levels in the blood has a negative feedback effect on further release of corticotropin from the hypothalamus. An excess of corticosteroids may thus be caused by an ACTH-secreting pituitary adenoma or an adrenal cortex adenoma, or an ectopic ACTHsecreting tumor. Usually, magnetic resonance imaging scans of the brain will show an enlarged pituitary gland if the location of the tumor is in the anterior pituitary gland. However, in some patients the precise location of the adenoma is unclear. It is thus necessary to stimulate secretion of the pituitary gland by corticorelin and sample cavernous sinus and peripheral venous blood samples for levels of the ACTH. 1'2 The procedure is fairly simple. Using biplane angiography, bilateral femoral venous catheters are inserted. The microcatheters are advanced bilaterally into each cavernous sinus. Baseline blood samples from the periphery and cavernous sinus are sent as indicated in Fig 1. Corticorelin is then injected intravenously, and samples of blood are taken bilaterally from each of the areas as indicated at intervals of 5, 10, 15, and 20 minutes (Fig 2). The anesthesia considerations for this procedure are dictated by the pathophysiology of Cushing's disease. These patients are often hypertensive and have elevated serum glucose levels. Care must be taken in positioning the patient and protecting their extremities because they have such fragile tissues. Their obesity may make airway management, intubation, and securing intravenous access very difficult. The procedure may be done under local or general anesthesia. Our interventional neuroradiologist usually prefers general anesthesia for these cases unless there are good reasons to do this with intravenous sedation plus local anesthesia. The procedure is done on an out-patient basis, and there should be minimal post-procedure pain or complications.

Lack of Effect of Corticosterone Administration on Male Sexual Behavior of Rats

RETANA-MARQUEZ, S., H. BONILLA-JAIME AND J. VELAZQUEZ-MOCTEZUMA. Lack of effect of corticosterone administration on male sexual behavior of rats. PHYSIOL BEHAV 63(3) 367–370, 1998.—The increase in plasma levels of corticosteroids as part of the stress response has been associated with failure in the reproductive function in most vertebrate species, both in females and males. Recently, we have shown that male sexual performance in rats is readily affected by different stressors, both acutely and chronically applied. However, there are few reports that directly correlate the increase in corticosteroid levels with the behavioral effects of stress. In this study we investigated whether the administration of corticosterone, either acutely or chronically, could reproduce the effects of stress on male sexual behavior in the male rat. Four doses of corticosterone (0.5, 1, 2, and 4 mg) or the vehicle, were administered during four consecutive days to sexually experienced males. Male sexual behavior was assessed after the first and the fourth injection. After the last test, males were killed and levels of corticosterone and testosterone were measured by HPLC. We observed an increase in corticosterone plasma levels in a dose-dependent manner. None of the sexual behavior parameters, however, was modified. Plasma levels of testosterone were not modified by corticosterone administration. Both steroids were increased in response to sexual activity, though. These data show that, unlike amphibians and female mammals, corticosteroids do not alter sexual behavior in male rats and suggest that the effect of stress on male sexual behavior cannot be explained by increases in corticosterone.

Corticosteroid Regulation of Gene Expression and Binding Characteristics of Vasopressin Receptors in the Rat Brain

Arginine-vasopressin (AVP) plays significant roles in neuroendocrine and autonomic regulation, and in processing of cognitive information. Its synthesis and secretion are subject to control by circulating glucocorticoids. The lateral septum and subdivisions of the hippocampus are innervated by AVP-ergic fibres and, together with AVP-producing neurons in the hypothalamic paraventricular nucleus, are major neural targets of glucocorticoid negative feedback. In this study, we investigated the effects of chronic adrenalectomy (ADX) and subsequent treatment with supraphysiological doses of corticosterone (B) on the gene expression of AVP receptors of the V1a subtype in the septum, hippocampus and hypothalamic arcuate (ARC) nucleus using semiquantitative in situ hybridization histochemistry. Adrenalectomy did not alter AVP receptor expression in any of the structures studied. Supplementation with B significantly decreased AVP receptor expression in the lateral septum and hippocampus, whereas receptor mRNA levels in the ARC were indistinguishable from those measured in controls. In a complementary study, we investigated the binding characteristics of V1 AVP receptors in membrane preparations from the hippocampus. Adrenalectomy significantly decreased the number of AVP binding sites, and chronic corticosteroid treatment was associated with a further suppression of AVP receptor concentrations in this structure. These results indicate that the gene transcription of V1a AVP receptors in the brain is regulated by circulating glucocorticoids in a site-specific fashion that largely reflects the corticosteroid sensitivity of the corresponding structure.(ABSTRACT TRUNCATED AT 250 WORDS)

Hematopoietic effects in mice exposed to deltamethrin and hydrocortisone

The effects of deltamethrin on bone marrow and spleen progenitor cell responsiveness to granulocyte and macrophage colony-stimulating factors were evaluated. Delthamethrin was tested in parallel with hydrocortisone to further investigate some similarity in the in vivo effects of both compounds observed in previous studies in our laboratory. In vivo effects were studied after the subcutaneous administration in mice of three 5 mg/kg injections of deltamethrin and a single 30 mg/kg injection of hydrocortisone to Balb/c mice. Soft agar colony formation, marrow andspleen cell counts, body, spleen and thymus weights were determined. Data obtaied in vivo indicate that deltamethrin and hydrocortisone reversibly icrease the formation of granulocyte an macrophage colonies in the narrow, but not in the spleen. No changes were observed in total and differential cell counts in the marrow and spleen and spleen weights. Treatment with both compounds, however, resulted in a dramaic reduction in thymus weights. Assays for endotoxin demonstrate that these effects were not due to the liberation of endotoxin. In vitro addition of 10 −5, 10 −2 10 −7 M deltamethrin and hydrocortisone to marrow cultures from untreated mice resulted in different effects from those observed in vivo. Hydrocortisone increased granulocyte and reduced macrophage colonies, whereas deltamethrin was without in vitro effects. It is suggested that deltamethrin effects are due to an indirect action on the hypothalamic - pituitary axis leading to increased corticosteroid levels. The importance of biotransformation mechanims is also emphasized.

Morphine inhibits signal transduction subsequent to activation of protein kinase C in mouse splenocytes

Morphine administered as a subcutaneous implant was previously reported to inhibit the mitogen-induced initial increases in cytoplasmic free calcium concentrations ([Ca 2+]i) in mouse splenocytes. The present studies were initiated to determine whether morphine affects signal transduction subsequent to activation of protein kinase C (PKC) in immune cells. Administration of morphine significantly inhibited the phorbol myristate acetate (PMA)-stimulated increase in interleukin-2 receptor (IL-2R) expression in both CD4+ and CD8+ mouse T cells. In contrast, morphine treatment had no effect on PMA/calcium ionophore (A23187)-induced increase in IL-2 secretion. suggesting a selective inhibition of IL-2R expression. Simultaneous administration of morphine and the opiate antagonist naltrexone blocked the effect of morphine on CD4+ cells. The inhibition of PMA-stimulated IL-2R expression was not reproduced by incubating splenocytes with morphine (10 −8–10 −5 M). These results suggest that this effect of morphine was mediated through opiate-receptors, but not directly via opiate receptors located on T cells. Moreover, adrenalectomy abolished this effect of morphine in CD4+ but not CD8+ T cells, suggesting that the inhibitory effect of morphine on IL-2R expression in CD4+ T cells may be mediated through a morphine-induced increase in corticosteroid levels. Thus, opiate-induced immunosuppression may involve an inhibition of post-PKC events, especially IL-2R expression, as well as impairment of earlier events in the activation of immune cells such as the increase in [Ca 2+]i.

Corticosteroid and immune responses of mice following mini-osmotic pump implantation

Serum corticosteroid (CS) levels were measured in (B6xA)F1 mice following the implantation of ALZET mini-osmotic pumps. Studies of shipment stress were used as a basis for comparison. At 24 and 48 h following the implantation of pumps CS levels were moderately elevated. However, immunization of animals at 24 h postsurgery with sheep erythrocytes showed normal immune responses. The results of these experiments indicate that the use of mini-osmotic pumps for the delivery of immunomodulatory drugs induces moderate and short-term increases in CS levels, which do not appear to interfere with normal immune function.

Relationship of the hypothalamic-pituitary-adrenal axis with chemically induced immunomodulation. I. Stress-like response after exposure to T-2 toxin

T-2 toxin (T-2), a trichothecene mycotoxin, produced by several members of the genus Fusarium is a cytotoxic feed contaminant and has been shown to by immunomodulatory. It is suspected that T-2 associated immunomodulation is mediated partly through the hypothalamic-pituitary-adrenal (HPA) axis. T-2, prepared in 4% ethanol/corn oil, was administered orally to male CD-1 mice. Endotoxemia was evident 24 h after a single, oral exposure to T-2. Blood levels of corticosterone, indicative of the stress-response, increased 24 h after T-2 exposure. Hypothalamic norepinephrine and serum corticosterone levels increased in a dose-related manner after 2 weeks of T-2 exposure. Endotoxin, detected in the serum of animals exposed to 2.5 mg/kg T-2 for 1 week, was not associated with bacteremia. Neither endotoxin nor bacteremia were detectable after 2 or 4 weeks of T-2 exposure. Exposure to 2.5 mg/kg T-2 also affected several organs. The forestomach was ulcerated with lymphocytic infiltration, epithelial proliferation, and hyperkeratinization. Increased spleen weight was associated with a proliferative red pulp. No histological changes were observed in the enlarged liver. Gastritis has been associated with increased corticosteroid production; cortical depletion and reduced mass of the thymus are phenomena attributable to increased corticosteroid levels. An increased corticosteroid level has been associated with thymic involution leading potentially to decreased T-dependent antibody response, a known effect of T-2.

The effect of heat-stable extract from bovine sublingual salivary glands on the carbohydrate metabolism of the rabbit

This extract (SLF 3) caused hyperglycaemia when injected intraperitoneally (i.p.) into rabbits (20 mg/kg body wt) which was maximal at 0.5 h and took 2 h to return to the initial value. SLF 3 also produced a two-fold increase in corticosteroid levels compared to controls, so the increase in blood glucose was probably mediated by adrenal cortical hormone stimulation.

Opiate effects on plasma corticosteriods: relationship to dysphoria and self-administration

Narcotic analgesics have been shown to raise plasma corticosteroids in the rat. This effect has been interpreted as a response of the rat to the dysphoric properties of the drug. This interpretation is based on the observation that drugs such as nalorphine or cyclazocine which cause dysphoria in man elevate plasma corticosteroids in the rat at relatively low doses. Drugs like morphine or pentazocine which induce little dysphoria in man elevate plasma corticosteroids in the rat only at much larger doses. The corticosteroid-elevating effect is mediated by an opiate receptor since naloxone antagonizes the effect of morphine or the analgesic, U-50,488. Those narcotic analgesics which increased corticosteroid levels at low doses were also found to be poorly self-administered by rats. In contrast, compounds like morphine, pentazocine and the analgesic, U-49,274A, which were self-admistered at high rates, elevated corticosteroids only after large doses. The relationship between the dose of a drug which causes elevations in corticosteroid levels and whether or not the drug is self-administered further supports the premise that elevated corticosteroid levels induced by analgesics is due to their dysphoric properties.

HORMONAL EFFECTS ON RAT MAMMARY GLAND IN VITRO

The mammogenic and lactogenic effect of various hormones on rat mammary gland has been studied in vitro using an organ culture technique. The effects have been assessed mainly by light microscopy. The study has been confined mainly to the effects and interactions of the lactogenic protein hormones, corticosteroids and progesterone. It establishes the importance of the presence of insulin and serum and of the type of serum for the hormonal effects in vitro. These effects have been studied mainly on the explants obtained from 13 days pregnant rats. In addition explants from non-pregnant rats and at various stages of pregnancy and lactation were studied.

Protein binding of plasma cortisol in the foetal lamb near term.

Binding of cortisol to plasma proteins was studied in the foetal lamb by equilibrium dialysis at 37 degrees C. At 122 days of pregnancy the mean level of transcortin expressed as cortisol-binding capacity was 28 +/- 6 (S.D.) ng cortisol/ml plasma. During the last 14 days of pregnancy there was a progressive increase in transcortin-binding capacity to 85 +/- 14 ng cortisol/ml plasma. A sharp increase in the concentration of both protein-bound and unbound cortisol was observed over the same period. A rise in the concentration of total cortisol from around 3 to 42 ng/ml was associated with an increase in unbound cortisol from 0-2 to a maximum of 2-1 ng/ml. The concentration of albumin-bound cortisol was approximately equal to that of unbound cortisol. The mean value for the transcortin-cortisol affinity constant was 1-15 x 10(8) l/mol. It is concluded that an increase in transcortin-binding capacity is partly responsible for the prepartum increase of corticosteroid levels observed in normal foetal lambs.