RESUMEN
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.
Asunto(s)
Dihidrotestosterona/sangre , Sueros Inmunes/aislamiento & purificación , Radioinmunoensayo/métodos , Testosterona/sangre , Animales , Afinidad de Anticuerpos , Especificidad de Anticuerpos , Dihidrotestosterona/inmunología , Adyuvante de Freund/administración & dosificación , Sueros Inmunes/análisis , Inmunización/métodos , Radioisótopos de Yodo , Masculino , Conejos , Radioinmunoensayo/instrumentación , Testosterona/inmunología , Factores de TiempoRESUMEN
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.
Asunto(s)
Dihidrotestosterona/sangre , Radioisótopos de Yodo , Radioinmunoensayo/métodos , Testosterona/sangre , Animales , Cromatografía en Capa Delgada , Dihidrotestosterona/inmunología , Marcaje Isotópico/métodos , Masculino , Conejos , Radioinmunoensayo/instrumentación , Testosterona/inmunología , Testosterona/aislamiento & purificaciónRESUMEN
Testosterone-17-hemisuceinate (T-17-HS) by esterification with succinic anhydride technique and Testosterone-3-carboxy-methyl-oxyme (T-3-CMO) derivatives were synthesized. The T-protein (BSA) conjugates were prepared by the condensation with carbodiimide technique, for the T-17-HS derivative and by the mixed anhydride technique, for the T-3-CMO. Both the derivatives and the T-BSA conjugates were characterized by melting point, ultraviolet and infrared spectra, thin layer chromatography. The physico-chemical characteristics and the hapten-protein incorporation being convenient: 28 mol T/mol BSA for T-17-HS-BSA and 38 mol T/mol BSA for T-3-CMO-BSA, the T-carrier conjugates are adequate for their use as immunogens.
Asunto(s)
Dihidrotestosterona/química , Radioinmunoensayo/métodos , Testosterona/análogos & derivados , Testosterona/química , Animales , Cromatografía en Capa Delgada , Humanos , Espectrofotometría Ultravioleta , Testosterona/análisis , Testosterona/síntesis químicaRESUMEN
The hypophyseal human growth hormone (hGH), a Raben type laboratory preparation, was re-evaluated as regards its innocuity for therapeutic use. Besides the usual control tests recommended by the Romanian Pharmacopoeia, the contamination of the hGH for clinical use with acute and slow viruses, was investigated taking into account the withdrawal of this hormone in many developed countries. The contamination was absent both with acute viruses as resulted from hemadsorption on cell cultures and counterimmunoelectrophoresis, and with slow viruses as observed from a two year-follow up of guinea pigs injected intracerebrally with the hGH preparation. Further, the content of the growth hormone itself as well as the contamination degree with other pituitary hormones was examined. The hGH-RIA content was 2.23 +/- 0.13 IU/mg (means +/- SEM), range: 1.38-2.80 IU/mg (1st International Standard hGH 80/505-1982). The prolactin contamination assessed by RIA was 187.34 +/- 37.66 ng/mg hGH, range: 28.44-385.20 ng (International Standard WHO: 80/541). The LH and FSH contamination as quantified by the Isocommerz (DDR) RIA kits was with two orders of magnitude lower than 10 IU-LH/IU-hGH, the upper LH contamination limit considered as acceptable. Moreover, the proportion of the large molecular forms in the lyophilized hGH preparation was investigated by polyacrylamide gel electrophoresis. Corroborating the data obtained by these control tests with our previous experience on 149 pituitary dwarfs treated with this hGH preparation during the interval 1964-1984, resulted minor risks of some dangerous side effects of hGH administration in children with growth hormone deficiency by possible contamination with pathogenic agents or with other disturbing hormones.
Asunto(s)
Contaminación de Medicamentos , Trastornos del Crecimiento/tratamiento farmacológico , Hormona del Crecimiento/efectos adversos , Niño , Preescolar , Contrainmunoelectroforesis , Electroforesis en Gel de Poliacrilamida , Hormona Folículo Estimulante/análisis , Hormona del Crecimiento/análisis , Hormona del Crecimiento/normas , Humanos , Hormona Luteinizante/análisis , Prolactina/análisis , RadioinmunoensayoRESUMEN
Plasma testosterone (T), 5 alpha-dihydrotestosterone (DHT), T/DHT ratio, estrone (E1), estradiol (E2), E2/T ratio and 17-hydroxyprogesterone (17-OH-P) were measured in 70 patients with prostatic carcinoma (PCA) (T34, N01, M01), after endocrine therapy including: steroidal estrogens (polyestradiol phosphate = Estradurin monthly in injections of 80 mg; estradiol - 17 beta: 5 mg/d), nonsteroidal synthetic estrogens (DES: 5 mg/d; chlorotrianisene = TACE: 24 mg/d; DES-diphosphate = Honvan: 360 mg/d), orchidectomy and their combinations. All these forms of treatment lowered T, DHT, T/DHT ratio and 17-OH-P, the maximal suppression being observed after castration. No further decrease of the mentioned parameters was achieved by estrogen treatment of castrated patients, as compared with castration alone. In the noncastrated patients, steroidal estrogens were less effective than nonsteroidal ones, in terms of lowering T, DHT and T/DHT ratio. Significant decreases in estrone (p less than 0.05) and estradiol (p less than 0.01) levels were observed after nonsteroidal estrogen treatment and castration respectively, and extremely high values of circulating estrogens were found after steroidal estrogen administration.
Asunto(s)
Congéneres del Estradiol/uso terapéutico , Neoplasias de la Próstata/tratamiento farmacológico , Esteroides/sangre , 17-alfa-Hidroxiprogesterona , Anciano , Castración , Dihidrotestosterona/sangre , Estradiol/sangre , Estrona/sangre , Humanos , Hidroxiprogesteronas/sangre , Masculino , Persona de Mediana Edad , Neoplasias de la Próstata/sangre , Testosterona/sangre , Factores de TiempoRESUMEN
Estradiol receptor (ER) and progesterone receptor (PR) content along with the cytosol and plasma estrone and estradiol levels in 15 premenopausal and 26 postmenopausal women with breast cancer in different clinical stages (T123, N01, M0) were measured. ER-positive tumor frequency and the ER content tended to be higher in postmenopausal than in premenopausal patients. There was no evidence for a relationship between high cytosol estrogen levels and low receptor measurements. The estrogen concentration was higher in ER-positive tumor cytosols than in those of ER-negative tumors; the differences were significant in postmenopausal women, only, with P less than 0.05 for estrone and P less than 0.01 for estradiol values. Twelve pairs of tumor and normal tissue from the same breast, were also studied: seven of which contained ER-positive and five ER-negative tumors. The ER-positive tumors showed a clear trend to higher estradiol content as compared to the corresponding normal tissues. The circulating level of estradiol in postmenopausal women, was higher (P less than 0.05) in ER-positive tumors than that in ER-negative tumors. Our results indicate that: (a) false negative ER assays are not likely to be due to the presence of endogenous estrogens. (b) higher amounts of estrone and estradiol are contained in ER-positive tumors than in negative ones.
Asunto(s)
Adenocarcinoma/metabolismo , Neoplasias de la Mama/metabolismo , Mama/metabolismo , Estradiol/metabolismo , Estrona/metabolismo , Receptores de Estrógenos/metabolismo , Receptores de Progesterona/metabolismo , Citosol/metabolismo , Femenino , Humanos , Cinética , Menopausia , Estadificación de Neoplasias , Receptores de Estradiol , Valores de ReferenciaRESUMEN
Plasma cortisol, 17-hydroxyprogesterone (17-OH-P), testosterone (T), 5 alpha-dihydrotestosterone (DHT, estrone (E1) and estradiol (E2), were measured in 94 normal adult men aged between 20-99, using RIA methods after chromatographic separation of steroids on Sephadex LH-20 columns. All plasma steroids except 17-OH-P, were age dependent: cortisol, testosterone and DHT decreased significantly with age, whereas estrone and estradiol were significantly increased in elderly men. Cortisol, testosterone, T/DHT ratio and estradiol levels were significantly correlated with age. The age related changes of plasma steroids in elderly men, were suggestive of decreased cortisol secretion, and decreased testicular function with increased peripheral conversion of androgens into estrogens. Testosterone was positively correlated with its precursor (17-OH-P) and respectively its peripheral metabolites (DHT and E2). The negative correlation between estrone and 17-OH-P found in elderly men, suggested that increased estrogen level in aging males may be considered able to inhibit the testicular androgen production.
Asunto(s)
Dihidrotestosterona/sangre , Estradiol/sangre , Estrona/sangre , Hidrocortisona/sangre , Hidroxiprogesteronas/sangre , Testosterona/sangre , 17-alfa-Hidroxiprogesterona , Adulto , Anciano , Envejecimiento , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Cortisol, 17-hydroxyprogesterone (17-OH-P), progesterone, testosterone, 5 alpha-dihydrotestosterone (DHT), estrone (E1) and estradiol (E2) were determined by RIA after chromatographic separation of steroids on Sephadex LH-20 columns, in 54 hospitalized patients with benign prostatic hyperplasia (BPH) and in 32 hospitalized patients with prostatic carcinoma (PCA) (T34, N01, M01). The patients' values were compared with those of 63 age-matched controls. Increased cortisol and DHT levels, subnormal estrogen and 17-OH-P values and normal progesterone level were found in both benign and malignant groups. Higher mean values for testosterone, and T/DHT ratio and lower mean values for E2/T ratio were found in PCA, as compared with those in BPH. An age invariance of cortisol, testosterone, T/DHT ratio and estradiol was found in both BPH and PCA, instead of the age dependence found in normal subjects. The normal relations between testosterone and its precursor (17-OH-P) and its peripheral metabolite (E2), respectively, and the normal relation between estrone and 17-OH-P were not evident in either BPH or PCA group. The normal direct relation between testosterone and DHT has been found in both patient groups.
Asunto(s)
Dihidrotestosterona/sangre , Estradiol/sangre , Estrona/sangre , Hidrocortisona/sangre , Hidroxiprogesteronas/sangre , Progesterona/sangre , Hiperplasia Prostática/sangre , Neoplasias de la Próstata/sangre , Testosterona/sangre , 17-alfa-Hidroxiprogesterona , Factores de Edad , Anciano , Humanos , Masculino , Persona de Mediana Edad , Valores de ReferenciaRESUMEN
The authors have investigated the presence of oestrogenic and progesteronic receptors in 38 patients with breast cancer. It was noted that these receptors were present in 20 patients, and that the hormone-dependent tumours are more frequent in patients which are in the post-menopausal stage. The level of circulating oestrogens was also investigated, as well as the amount of oestrogens in the cytosol of tumor cells, and in the normal cells from the diseased breast. In patients with oestrogen-positive tumors, and/or progesteron-positive tumors, a rise was noted in the amount of circulating oestradiol, an increased contents of cytosolic oestradiol, especially in the tumoral cells as compared with the normal ones in the same breast. On the basis of these data patients were selected, with hormone-dependent breast tumors in view of ovarectomy. The indications are discussed, as well as the advantages of surgical ovarectomy.