W.R. Robertson

Antibody that blocks stimulation of cortisol secretion by adrenocorticotrophic hormone in Addison's disease

To investigate whether Addisons disease may in some cases be due to the blocking of adrenocorticotrophic hormones action at the adrenal cortex by antibodies IgG isolated from a woman with Addisons disease associated with the autoimmune polyglandular syndrome type I was studied. Its effects on guinea pig adrenal cells in vitro were investigated and compared with those of IgG from three normal subjects and IgG obtained commercially. IgG from the patient inhibited the stimulation of cortisol secretion by adrenocorticotrophic hormone by 77 (SD 2)% and 57 (12)% at concentrations of 0·5 and 0·05 g/l, respectively; IgG prepared five months after she had started treatment with replacement steroids inhibited cortisol secretion by 74 (1)% (0·5 g/l) and 51 (15)% (0·05 g/l). The other IgGs had no inhibitory effects. The IgG from the patient and that obtained commercially did not inhibit the stimulation of cortisol secretion by dibutyryl cyclic adenosine monophosphate or precursors of cortisol. None of the IgGs bound to adrenocorticotrophic hormone. These results suggest that the IgG from the patient acted against the receptor for adrenocorticotrophic hormone, and its presence may explain the patients raised concentrations of adrenocorticotrophic hormone, failure to respond to exogenous adrenocorticotrophic hormone, and normal basal cortisol concentrations. Addisons disease may thus in some instances be a receptor antibody disease.

Human chorionic gonadotrophin contributes to the bioactivity of Pergonal

OBJECTIVE We examined batch variation in the LH‐like bioactive content of Pergonal and determined whether hCG contributes to this.

The effect of ketoconazole on adrenal and testicular steroidogenesis in vitro

The effect of the anti-fungal agent, ketoconazole, on cortisol secretion from adrenal cells stimulated with ACTH (1-24, 50 ng/l) and on testosterone secretion from Leydig cells stimulated with LH (5 i.u./l), has been tested. The concentration of drug which inhibited cortisol and testosterone secretion by 50% (ED50) was 3.6 +/- 0.7 mumol/l and 0.61 +/- 0.03 mumol/l, respectively. The effect of ketoconazole on adrenal and testicular steroidogenesis was completely reversible. Thus, adrenal and testicular cells which had been washed after exposure to greater than 95% inhibitory dose of ketoconazole responded in a similar manner to hormone stimulation as cells similarly washed and which had not been exposed to drug. The sites of the anti-steroidogenic effect of ketoconazole have been established using a method based upon the sequential stimulation by the exogenous precursor steroids of the various steps leading to the biosynthesis of cortisol by adrenal cells and testosterone by Leydig cells. We conclude that ketoconazole reversibly inhibits the sequence between ACTH/LH binding and pregnenolone production and also inhibits testicular C-17-C-20, lyase activity.

Assessment of the biopotency of anti-thyroid drugs using porcine thyroid cells

The effect of six drugs on the uptake and organification of iodide by porcine thyroid cells stimulated with bovine TSH (10 miU/L) has been investigated. The drugs fall into two categories: the peroxidase inhibitors, methimazole (MMI), 2-thiouracil (2-TU) and 3-amino 1,2,4 triazole (3-ATA) and the ionic inhibitors, lithium chloride (LiCl), potassium perchlorate (KC10(4], and sodium iodide (NaI). All the drugs led to a dose-related inhibition of iodide metabolism. The most potent effect on iodide uptake was seen with NaI which inhibited this function by 20% even at 10(-8) mol/l. In contrast, the most potent effect on iodide organification was observed with methimazole which led to a 25% inhibition at 10(-8) mol/l. The concentrations of drug which gave rise to a 50% inhibition of iodide uptake were (mumol/l) 0.26 (NaI), 3.5 (KClO4), 9.7 (2-TU), 15 (MMI), 26 (3-ATA), and 1500 (LiCl). The comparable figures for organification were 0.13 (MMI), 0.18 (2-TU), 0.23 (NaI), 1.2 (3-ATA), 15 (KClO4) and 1300 (LiCl). We conclude that this in vitro system has considerable potential for the assessment of potency and possible bioassay of anti-thyroid drugs of varying structures and sites of action.

Physiological oestradiol administration does not change the biological to immunological ratio of LH in serum from post‐menopausal women

OBJECTIVE We Investigated the effect of chronic physiological concentrations of oestradiol on the bioactivity (B) and immunoactivity (I) of LH in serum using radioimmunoassay (RIA) and a sensitive Immunoradlometrlc (IRMA) assay.

Assessment of the Bioactivity of Human and Bovine Thyrotrophin Preparations Using a Porcine Thyroid Cell Bioassay

The biopotency of six preparations of thyrotrophin (TSH) has been compared in a highly sensitive in vitro porcine thyroid cell bioassay using iodide uptake as an endpoint. Three of these preparations were of human origin and three derived from bovine pituitary tissue. One human TSH preparation, the 2nd International Reference Preparation, 80/558, was used to calibrate the other five. The log dose—log response curves for all preparations were sigmoidal in shape. For the purpose of evaluation the central linear portions of the curves were compared. With all preparations the slopes in this region were very similar. The relative biopotencies of the bovine preparations (unit : unit) were at least five times those of the human standards when measured using the porcine thyroid cell bioassay. These findings emphasise the need to control the TSH standards employed in a variety of bioassays, particularly those used for between-laboratory comparison.

Hepatic and Endocrine Effects of Azole Antifungal Agents

The group of drugs known collectively as the azoles, comprising a number of 1-substituted imidazole and triazole compounds, undoubtedly represents the modern approach to both topical and systemic treatment of fungal disease. The first generation of azole antifungals, lipophilic imidazole compounds such as clotrimazole, econazole and miconazole, exhibited poor systemic availability following oral administration, due to both poor absorption and ex-tensive first-pass metabolism. Their use has consequently been essentially limited to topical treatment of superficial fungal infections. Ketoconazole, a more polar imidazole introduced into therapy in the late 1970s (Thienpont et al. 1979), represented a breakthrough in the treatment of antifungal disease since it was the first imidazole agent with good oral bioavailability in humans (Heel et al. 1982). Consequently, it was introduced for the treatment of both superficial and systemic fungal infections by oral administration. The subsequent extensive systemic use of ketoconazole resulted in the appearance of two major classes of side effect: drug-related idiosyncratic hepatitis and decreased plasma concentrations of steroid hormones. In addition, and some-what less serious in impact, the drug has also been shown to interfere with the metabolism of a number of co-administered drugs.

Differences in serum luteinizing hormone measurements by immunoradiometric assay induced by kinetic manipulation of assay conditions are dependent on the endocrine milieu of serum.

Divergent estimates for luteinizing hormone (LH) in individual serum samples may be given by different immunoassays. In order to investigate this phenomenom further, we have studied the effect of differences in assay kinetics within the same immunoradiometric assay (IRMA) configuration on LH measurement in sera from different endocrine states. Three pairs of monoclonal/polyclonal two-site IRMA systems for LH were developed from three LH monoclonal antibodies and a common polyclonal anti-human chorionic gonadotrophin. For IRMA systems a short and long assay, which were different only with respect to the incubation time (1/2 h and overnight respectively), of the labelled monoclonal first antibody were performed. The IRMAs were all standardized against the LH international reference preparation 68/40. LH concentrations were measured by all the IRMAs in sera obtained from normal men (n = 11) and from women with polycystic ovarian syndrome (PCO; n = 13). In normal men, there were no differences in LH estimates between the short and the long assays of the three IRMA systems, and the ratios of long to short assays were similar for all the systems. However, in PCO there were significant differences between short and long assays and the ratios of long to short assays were different for the IRMA systems. These results indicate that kinetic differences between IRMAs of the same antibody configuration can be associated with differences in measured LH concentrations, depending on the endocrine status of the sera studied. As LH glycoform patterns are known to differ between normal men and PCO, the observed changes in LH estimates may be due to the different glycoform composition.