The development of a radioimmunoassay system for testosterone (T) and dihydrotestosterone (DHT). Part 2. The preparation of antisera to T.

Testosterone-3-O-carboxymethyl-oxime derivative was synthetized and coupled to bovine serum albumin (BSA). The T-3-CMO-BSA conjugate homogenized with Freund's adjuvant used as immunogen was injected multiple sites in rabbits. The antisera collected were characterized in a radioimmunological system, separation with dextran-charcoal using 125I-Testosterone as tracer. The antibody titres varied from one animal to another. The titre of anti-T serum selected for RIA was 1: 10(4)-1: 2 X 10(4) (initial dilution). All anti-T sera 100 percent crossreacted with 5 alpha-dihydrotestosterone but nonsignificant interference was observed with other C19, C21 and C18 steroids. The affinity constant of the selected anti-T serum was in the range 1.4-1.9 X 10(9) litres/mole. The data so far published on the antisera toward testosterone are reviewed. We conclude that the selected anti-T-3-CMO-BSA serum may provide assays for testosterone with potential for clinical applications.

The development of a radioimmunoassay system for testosterone (T) and dihydrotestosterone (DHT). Part 3. The preparation of radioiodinated testosterone.

A two step-method was applied for the preparation of a tracer adequate for a radioimmunoassay (RIA) system for testosterone. Histamine was radioiodinated by the Chloramine-T method and then coupled to testosterone-3-carboxymethyl-oxime (T-3-CMO) derivative. After purification by TLC, the steroid tracer was stable in ethanol for at least four months. Using an anti-T-3-CMO-BSA serum, a (bridge) homologous RIA system for testosterone was developed. The reagents (antiserum, tracer and standard) were incubated 2 hrs at 37 degrees C and then the free radioactivity was removed by a dextran-charcoal suspension. The testosterone RIA system, with the sensitivity of 30 pg/tube, is suitable for the measurement of the steroid hormone in biological fluids and tissues.

Unusually large adrenal adenoma excreting unusually large amounts of androgen metabolites in urine.

A 27-yr female patient was admitted to the hospital because of secondary amenorrhea and a severe and complex virilization syndrome. The urinary 17-Ks, 17 OHCS and testosterone glucuronide values were greatly increased. The plasma testosterone through very high, was within the normal limits in males (3.52 ng/ml). Urography, echography and CT scan revealed a large right adrenal tumor. Removal of the tumor brought about a considerable clinical improvement: reoccurrence of the menstrual cycle, a progressive decrease in the virilization syndrome and normalization of the urinary 17-Ks, 17-OHCS and of the plasma testosterone.

Effect of a melatonin-free extract from bovine pineal on plasma and urinary steroids and on serum prolactin in hirsute amenorrheic adolescents.

Basal and post-suppressive dexamethasone (dxm) levels of some urinary androgen metabolites, plasma testosterone (T), 17 hydroxyprogesterone (17 OHP) and of basal serum prolactin were determined in 34 hirsute amenorrheic adolescents aged 13-17 in whom a five day course of bovine pineal extract (4 ml a day i.m.) was instituted. No convincing effect of pineal extract administration on plasma T and 17 OHP and on serum prolactin was detectable in most of pineal-treated patients. Suppressibility to dxm of plasma T and 17 OHP was not a relevant index of both the origin of androgens or the clinical course of the patients.

Influence of the "Corynebacterium parvum" vaccine and stilbostat on intratesticular development of Walker 256 carcinosarcoma in hyper- or hypothyroidized Wistar rats.

This work studies the effect of nonspecific antitumoral immunity produced by the "Corynebacterium Parvum" vaccine (C.P.) and Stilbostal on the development of Walker 256 carcinosarcoma intratesticularly grafted to hyper- or hypothyroidized Wistar rats. The thyroid hormone level was altered by administration of thyroxin, Alkiron or surgical thyroidectomy. The C.P. vaccine was given in a mixture with tumoral cells suspension and the treatment with the Stilbostat drug was started 5 days from grafting. The results show that C.P. administered in a mixture with tumoral cells suspension causes marked inhibition of the development of the primary tumor and metastasis, an effect which is not significantly influenced by the thyroid hormone level. The antitumoral effect of the Stilbostat drug is enhanced by administration of C.P. in tumoral cells suspension. The greatest effect produced by Stilbostal was noted in the hyperthyroidized lots in which the tumor was administered in a mixture with C.P.

Influence of BCG on the development of 256 Walker carcinosarcoma inoculated intratesticularly in the rat, in relation to thyroid status alerations.

The effect of nonspecific antitumoral immunity caused by BCG on intratesticular development of 256 Walker carcinosarcoma has been studied on male hyper- or hypothyroidectomized rats. Thyroid hormones level was altered by administration of T4. Alkiron or thyroidectomy. The time and route of BCG administration differed from one lot to another. Authors' results show that administration of BCG before or concomitantly with the graft stimulates tumor development, whereas administration of BCG mixed with a suspension of tumoral cells causes marked inhibition in the development of primary tumor and metastasis, an inhibition which is not influenced by the thyroid hormones level. In vitro proliferation of the tumoral cells as well as histopathologic examination of the tumors suggest the involvement of a complex process in achieving the nonspecific antitumoral immunity.

BCG influence on 256 Walker carcinosarcoma growth in rats in relation to adrenal cortex status.

BCG influence on antitumoral immunity varied in relation to the timing and the way of administration, as well as in relation to the adrenal cortex status of the animals. BCG administration several days before tumor grafting, but especially when mixed with tumoral cells determined a maximum antitumoral effect, while vaccination after the tumor had been formed enhanced its metastasization. The decrease of the endogenous corticoids secretion amplified the antitumoral effect of BCG, while exogenous cortisone diminished this effect. The study of in vitro cultivated tumoral cells as well as microscopic examination of the tumors suggest the involvement of a complex process in the achievement of the nonspecific antitumoral immunity, a process in which the adrenal cortex status plays an important role.

Influence of stress and of endocrine imbalance on the experimental metastasis.

The influence of some stress agents applied during variable periods of time and under various modifications of the endocrine status was studied on the lymphotropic metastases of the 256 Walker carcinosarcoma, grafted intratesticulary in rats. Different reactivity of metastases, when compared to that of primary tumors was found in relation to the action of some compounds influencing the hormonal balance. Stress enhances the primary tumor development, while the metastatic growth is inhibited. However later on, sudden enhancement of the metastasis growth was observed. Adrenalectomy, as well as testosterone, thyroxine, of Stilbostat administration inhibit the metastasis, whereas cortisone, or antithyroid drug administration enhance the metastasis. The in vitro study of the tumor cell proliferation suggests some immunemorphological involvements in the development of this process.

Endocrine implications in experimental tumoral metastasis. III. The influence of the thyroid status on the metastasis of 256 Walker carcinosarcoma intratesticularly grafted and on the in vitro proliferation of the tumoral cells.

The influence of thyroid status modifications on the metastasis of 256 Walker carcinosarcoma intratesticularly grafted to rats was examined. Inhibition of metastasization and of the secondary tumoral growth was noticed in animals chronically treated with thyroxine, while hypothyroidization realized either by surgery or by administration of synthetical antithyroid drugs enhanced metastasization and growth of the secondary tumors. The primary tumoral development was not significantly influenced by the thyroid status modifications. The variations of the thyroid hormonal level influenced nonspecifically the proliferation of the in vitro cultivated tumoral cells.