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Biomedical subjects

R Scholler

Publications and source records attributed to R Scholler.

At least 55 records · Page 3Linked to original sources

Endometrium and plasma hormone profile in the peri-menopause and post-menopause.

Endometrial histology and plasma levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), 17 beta-oestradiol (E2), oestrone (E1) and progesterone (P) were studied in 483 women over a period of 13 yr (6 yr before and 7 yr after the start of definitive amenorrhoea, defined as the last menstrual bleeding). The patterns for these parameters were established on the basis of the results of 1227 gonadotrophin and steroid determinations and 721 endometrial biopsies. Three periods were identified. During the first, from year-6 to year-3, gonadotrophin levels increased gradually, while those of E2 remained normal, with peaks in some cases. Mean plasma P levels were within the normal range until year-3, but they then decreased progressively. Endometrial histology was similar to that observed during reproductive life. In the second period, from year-3 to year+1, there was a concomitant rise in gonadotrophins as the E2 and P levels decreased. However, at the start of definitive amenorrhoea, the mean E2 and P levels fluctuated between 60 and 100 pg/ml and between 2 and 3 ng/ml, respectively. The endometrium reflected this decrease in E2 and P production. It was not atrophic but proliferative when definitive amenorrhoea commenced. During the last period, from year+1 to year+7, gonadotrophins reached a plateau at high levels, while those of E2 continued to fall, reaching very low values at year+4, after which they reached a plateau. P levels were at the detection limit of the technique. The correlations between all plasma steroid levels and endometrial histology demonstrated discrepancies in 30% of cases: proliferative or hyperplastic endometria were seen at E2 levels of under 60 mg/ml, atrophic endometric at E2 levels of over 60 pg/ml and secretory endometria at very low P levels.

Atrophy↗

Steroid profiles during cycles with HCG stimulation of luteal function during in-vitro fertilization cycles.

178 in-vitro fertilization cycles have been explored for luteal function. Clomiphene and human menopausal gonadotrophins were used for ovarian stimulation and some patients were supplemented with human chorionic gonadotrophin (HCG) (5000 IU) on the day of laparoscopy, and also 2 and 4 days later. An imbalance of luteal function was observed in non-supplemented stimulated cycles, with hyperoestrogenicity and a relatively low production of progesterone. The administration of HCG reduced this imbalance by improving the production of progesterone without affecting the production of oestradiol-17 beta.

Adult↗

Regional heterogeneity of human spermatozoa detected with monoclonal antibodies.

The regional antigenic heterogeneity of human spermatozoa is confirmed with 6 monoclonal antibodies raised against ejaculated human spermatozoa. The topographical localization of the antigenic determinants suggests the existence of at least 6 domains on the human spermatozoon. Different fixatives had severe detrimental effects on the antigen-antibody binding. On live human spermatozoa, each antibody bound to a distinct region: acrosome, equatorial segment, entire tail, neck, midpiece and terminal piece. The antigens detected on the acrosome, equatorial segment and entire tail were surface components, whereas the other three were intracellular structures. The determinant present along the entire tail was a sperm-coating antigen. The molecular weights of the recognized antigens were estimated with the Western blot technique. Immunostaining of individual ejaculates established that the percentages of positive cells were 12-56% for the acrosome, 8-35% for the equatorial segment, 90-100% for the entire tail, 20-52% for the neck, 9-35% for the midpiece and 36-90% for the terminal piece. In addition, labelling of motile and immotile spermatozoa showed differences in the percentages of positive cells, with 5 out of 6 monoclonal antibodies, or in the fluorescence intensity, with the last one labelling the entire tail.

Antibodies, Monoclonal↗

[Adrenal reactivity to prolonged-action ACTH in hirsute women treated with 50 mg of cyproterone acetate combined with percutaneous estradiol].

A prolonged-action Synacthen (R) test (1 mg.i.m. was performed in 14 hirsute women before and 6 months after instituting combined treatment with cyproterone acetate (50 mg/day/21 days) and percutaneous estradiol (3 mg during the last 10 days of cyproterone acetate administration). Determination of blood cortisol and precursors of cortisol--plasma androgens--was carried out to evaluate possible modification of adrenal enzymatic activity. At the used doses, cyproterone acetate failed to influence adrenal reactivity to synthetic ACTH or the enzymatic activities of 21-hydroxylase and 17-beta-reductase in vivo.

Administration, Cutaneous↗

[Radioimmunoassay of 5-androstene-3 beta, 17 beta-diol in plasma and breast cyst fluid].

A simple and reliable radioimmunoassay for the determination of 5-androstene-3 beta,17 beta-diol (5-Adiol) in peripheral plasma and in breast cyst fluid, after chromatography on Celite microcolum has been described and evaluated. The antiserum used was raised in the rabbit injected with dehydroepiandrosterone-15 alpha-carboxymethyl-bovine serum albumin. In men below 40 years of age the levels ranged from 0.85 to 2.80 ng/ml (mean +/- SEM: 1.52 +/- 0.11; n = 24) and from 0.50 to 2.20 ng/ml (mean: 0.93 +/- 0.09; n = 20) in men aged between 41 and 62 years. The mean level was significantly different (p less than 0.001) between the two groups. A significant correlation (r = -0.56 p less than 0.01) was demonstrated between age and all male levels. In women the mean plasma level was in the follicular phase: 0.81 +/- 0.07 ng/ml (range: 0.40 - 1.50; n = 17; age: 19-41 years) and in the luteal phase: 0.83 +/- 0.05 ng/ml (range: 0.40 - 1.30; n = 29; age: 18-43 years). No cyclical change and no correlation with age could be evidenced. A significant difference (p less than 0.001) was shown between females and the young male group. In breast cyst fluid the levels of unconjugated 5-Adiol ranged from 0.05 to 13.70 ng/ml (mean: 2.21 +/- 0.73; n = 23) whereas the sulfate concentrations ranged from 75 to 7,500 ng/ml (mean: 1,973 +/- 543; n = 18), thus demonstrating very wide inter-individual variations.

Adult↗

Seasonal variations in plasma luteinizing hormone and testosterone levels in the European badger Meles meles L.

Although active spermatogenesis occurs throughout the year in the male European badger Meles meles L., seasonal fluctuations of testicular activity were observed. The present study compared the pattern of luteinizing hormone (LH) and testosterone (T) secretions throughout the annual cycle. LH secretion was investigated by means of a heterologous radioimmunoassay using anti-ovine LH beta antiserum and canine reference standard. Seasonal and nycthemeral changes in LH and T secretions were observed. During the year a period of maximal values for both hormones is well correlated with the breeding. Moreover, episodic LH releases occurs during the period of minimal testosterone values, indicating resumption of hypothalamo-pituitary activity well before the breeding period.

Animals↗

Cortisol, cortisone and dehydroepiandrosterone sulfate levels in umbilical cord and maternal plasma between 21 and 30 weeks of pregnancy.

Cortisol (F), Cortisone (E) and dehydroepiandrosterone sulfate (DHAS) were determined in maternal peripheral plasma and umbilical cord plasma between 21 and 30 weeks of amenorrhea. In fetal plasma, DHAS levels were the highest and those of F the lowest. E always exceeded F. The pattern of all these steroids was characterized by a plateau throughout the period considered. In maternal plasma, F levels were more elevated than those of E but lower than DHAS concentrations. All the steroids plateaued as in the fetus. The study of the correlation between the steroids in either the same milieu or in maternal and umbilical cord plasma demonstrated that fetal E was correlated with maternal F and with fetal F while fetal DHAS was inversely correlated with maternal E.

Cortisone↗

Plasma androgens during the luteal phase in a case of true hermaphroditism with bilateral ovotestis.

A true hermaphrodite with a bilateral ovotestis and a 46 XX karyotype was studied. This 14-year old subject developed ambiguous puberty with bilateral gynecomastia and stage IV public hair. Relatively high level of testosterone (T) (2.80 ng/ml), was found. The 5 alpha-reductase activity for T in the pubic skin was similar to that observed in normal adult males. A hemorrhagic corpus luteum in the left ovotestis was observed at laparotomy. The luteal phase immediately after dexamethasone administration (1 mg/day for 7 days) was characterized by a significant decrease of plasma androgens, T and androstenedione (A). The constantly low level of T (0.30 ng/ml) during the luteal phase in this subject did not appear to be due to the previously administered dexamethasone. This decrease of T production in the luteal phase might be secondary either to the increase of the estradiol-17 beta (E2) secreted by the corpus luteum or to the decrease of LH levels. Both mechanisms might act concomitantly.

Adolescent↗

Leydig cell tumor with gynecomastia: further studies--the recovery after unilateral orchidectomy.

The recovery of exocrine and endocrine testicular function was studied in six patients orchidectomized for an estrogen-producing Leydig cell tumor. Gynecomastia disappeared in four patients. The contralateral testis, whose volume was reduced, returned to normal size after 30 days. Sperm density returned to normal in only one of the four patients in whom the preoperative sperm count was reduced. One day after unilateral orchidectomy, plasma estradiol decreased to normal and testosterone (T) fell about 50%. On the 10th postoperative day, plasma T [5.60 +/- 1.20 ng/ml (SD)] was normal. On day 120, T was higher than on day 10 (6.83 +/- 1.20 ng/ml). There was no significant increase of T after a single injection of hCG (5000 IU) on day 10, and the T response was similar to that of normal men on day 120. Plasma FSH and LH were increased on the 10th postoperative day; they then decreased between 60 and 120 days after the operation but were still above the normal values on day 120. The FSH/LH ratio, which was 0.43 +/- 0.17 preoperatively returned to normal (1.60 +/- 0.25) 10 days postoperatively. In conclusion, after hemicastration for an estrogen-secreting tumor, testicular hormonal secretion returns to normal within 120 days but spermatogenesis may still be impaired at this time.

17-alpha-Hydroxyprogesterone↗

Seasonal variations of plasma prolactin and LH concentrations in the female blue fox (Alopex lagopus).

A heterologous radioimmunoassay system developed for the rabbit and suitable for a wide range of mammalian species has been shown to measure prolactin in the plasma of the blue fox. Evaluation of prolactin levels throughout the year showed the concentrations displayed a circannual rhythm with the highest values occurring in May and June. Prolactin concentrations remained low (approximately 2.5 ng/ml plasma) from July until April with no consistent changes found around oestrus (March-April). In 8 pregnant females, the prolactin increase in late April and May coincided with the last part of gestation and lactation: concentrations (mean +/- s.e.m.) increased to 6.3 +/- 0.6 ng/ml at mid-gestation, 9.7 +/- 2.1 ng/ml at the end of gestation and 26.7 +/- 5.0 ng/ml during lactation. In 10 non-pregnant animals, the mean +/- s.e.m. values were 7.2 +/- 1.2 ng/ml in April, 8.8 +/- 2.2 ng/ml in May and 9.8 +/- 1.3 ng/ml in June. The prolactin profile in 4 ovariectomized females was similar to that observed in non-pregnant animals, but the plasma values tended to be lower during the reproductive season (April-June). In intact females, the only large LH peak (average 28 ng/ml) was observed around oestrus. During pro-oestrus, baseline LH levels were interrupted by elevations of 3.1-10.4 ng/ml. During the rest of the year, basal levels were less than 3 ng/ml. In ovariectomized females, LH concentrations increased within 2 days of ovariectomy and remained high (35-55 ng/ml) at all times of year.

Animals↗

Seasonal variations of LH, prolactin, androstenedione, testosterone and testicular FSH binding in the male blue fox (Alopex lagopus).

The seasonal changes in testicular weight in the blue fox were associated with considerable variations in plasma concentrations of LH, prolactin, androstenedione and testosterone and in FSH-binding capacity of the testis. An increase in LH secretion and a 5-fold increase in FSH-binding capacity were observed during December and January, as testis weight increased rapidly. LH levels fell during March when testicular weight was maximal. Plasma androgen concentrations reached their peak values in the second half of March (androstenedione: 0.9 +/- 0.1 ng/ml: testosterone: 3.6 +/- 0.6 ng/ml). A small temporary increase in LH was seen in May and June after the breeding season as testicular weight declined rapidly before levels returned to the basal state (0.5-7 ng/ml) that lasted until December. There were clear seasonal variations in the androgenic response of the testis to LH challenge. Plasma prolactin concentrations (2-3 ng/ml) were basal from August until the end of March when levels rose steadily to reach peak values (up to 13 ng/ml) in May and June just before maximum daylength and temperature. The circannual variations in plasma prolactin after castration were indistinguishable from those in intact animals, but LH concentrations were higher than normal for at least 1 year after castration.

Androgens↗

Dissociation between testicular morphogenesis and functional differentiation of Leydig cells.

The aim of the study was to determine whether Leydig cells differentiate in vitro in gonads in which the formation of seminiferous cords is prevented by culture in a medium containing fetal calf serum. Appearance of 3 beta-hydroxysteroid dehydrogenase-positive cells and release of testosterone in the medium occurred at the same age irrespective of whether or not the gonads developed seminiferous cords. It is concluded therefore that testicular morphogenesis with the formation of seminiferous cords is not a prerequisite for the emergence and functional differentiation of Leydig cells.

3-Hydroxysteroid Dehydrogenases↗

[Comparison of conditions for immunodetection of antigens using monoclonal antibodies after electrotransfer on nitrocellulose membrane].

Solubilized human sperm proteins are separated by SDS-polyacrylamide gel electrophoresis under reducing conditions, electrophoretically blotted onto a nitrocellulose sheet and their antigenicity probed with 5 monoclonal antibodies. Bound proteins are exposed to different nonionic detergents and a comparative study is reported. The use of 0.05% Tween 20 or 0.05% Nonidet P-40 in the blocking and washing buffers and in the immunoprobes causes variations in the patterns of 3 monoclonal antibodies. Washing buffers containing either 0.1% of detergent (Tween 20, Nonidet P-40, Triton X-100) or different concentrations (0.05, 0.1, 0.4%) of Tween 20 remove prebound proteins from the nitrocellulose membrane. Nitrocellulose strips processed with different detergents lack different immunoreactive bands. The 5 monoclonal antibodies recognize degraded antigens and endogeneous proteolysis changes the antibody patterns.

Animals↗

Effects of freezing and thawing on mammalian oocyte.

In an attempt to study the deleterious effects which occur during the freezing and thawing of mammalian oocytes, we developed a cryomicroscope controlled by digital programmable equipment. The program permits any cooling rate between 0.1 and 60 degrees C/min with a precision of 0.6 degrees C. Using a precooled stage, it is possible to obtain rapid cooling (100 degrees C/min). The maximum thawing rate is about 60 degrees C/min. A copper-- constantan microthermocouple allows precise measurement of the specimen temperature. All information (specimen, temperature of the specimen, date, hour, and minutes) is recorded at the same time on photographic film by a camera fitted with a " Recordata Back" and a motor drive which allows three frames per second. Our preliminary results show that: (1) rapid cooling yields a supercooling with simultaneous extra- and intracellular crystallization; (2) slow cooling with seeding at -8 degrees C gives an extracellular crystallization which is achieved by -9 degrees C, followed by an extracellular recrystallization occurring at almost -8 degrees C which alters the morphology of the oocyte and the zona pellucida, without any visible intracellular crystallization; (3) during continued slow cooling the oocytes dehydrate without any intracellular freezing; and (4) during rewarming a partial rehydration of the cell occurs with a swelling of the oocytes to their original volumes after the thawing has been achieved.

Animals↗

Heterologous radioimmunoassay of fox LH: levels during the reproductive season and the anoestrus of the red fox (Vulpes vulpes L.).

A heterologous radioimmunoassay using ovine LH as the labeled hormone, canine LH as the standard, and an antiovine LH rabbit serum was validated for the measurement of fox LH. Physiological validation of the assay was evidenced by the high concentrations of LH at oestrus and following ovariectomy or the administration of LH-RH. Throughout the year, plasma LH levels demonstrate important variations, being low during and after the luteal phase (1.4 +/- 0.3 ng/ml) (mean +/- SE) and increasing during the second part of anoestrus (5.2 +/- 1.4 ng/ml). This latter increase might be correlated with that of androgens observed at the same period. Several LH rises preceded the preovulatory LH surge.

Androstenedione↗

Testicular secretion of conjugated and unconjugated steroids in normal adults and in patients with varicocele. Baseline levels and time-course response to hCG administration.

The effects of a single im injection of 10,000 IU of hCG on testicular steroid secretions were studied in 15 normal men and 54 patients with varicocele. Plasma levels of the following steroids were determined by radioimmunoassay: T, DHT, A, DHA, 5Adiol, E1, E2, 5P, 170H5P, 170HP, E1S, E2S, TE1, TE2, DHAS, 5AdiolS and hCG as well as FSH and LH in baseline samples. In normal men, peripheral venous blood was collected at 0, 1, 2, 4, 6, 8, 24, 36, 48, 72 and 96 h after hCG. Three types of steroid response were observed. The first, biphasic with an early peak within 2 h and a delayed one after 24 h, was shown by T, its unconjugated precursors and DHT. The second with an early rise plateauing for 6 h and followed by a 36 h peak characterized the estrogen pattern. Finally DHAS did not demonstrate a significant increase but 5AdiolS fluctuated at higher levels than the baseline ones. A significant correlation was only observed between hCG maximum value and E2 and TE1 maximum relative responses. In the patient group, peripheral blood was collected at 0, 1, 1.5, 2, 4, 6, 8, 10, 24 and 48 h after hCG. Baseline levels of all steroids were similar in the two groups. After hCG the same delayed pattern was observed as in the controls. The early relative response was low for T while A, DHT, E2 and E1S levels decreased. Conversely 170HP was higher until 24 h. Spermatic vein blood was collected before and at 1.5, 24 and 48 h after hCG. Steroid baseline levels in the varicocele side were similar to the other. After hCG, despite very large variations, general steroid patterns were comparable to those observed in peripheral vein blood. The important fall at 24 h of T/170HP and T/170H5P concomitant with that of T/E2 might be related to an inhibition of C17-20 lyase activity. The testicular secretion of all the measured steroids, including E1S, could be demonstrated except for DHAS. From our findings, a biogenetic defect evidenced by the fast response to hCG administration might exist in varicocele patients. In view of these results, the protocol of the hCG test should be improved using more frequent blood sampling and determining, besides T, its precursors and estrogen conjugates.

Adult↗

Estrogen 2-, 4-, 6- or 16-hydroxylation by human follicles shown by gas chromatography-mass spectrometry associated with stable isotope dilution.

Follicular fluid, obtained by aspiration of human Graafian follicles in cycles stimulated by clomiphene and hMG + hCG, was analyzed for estrogen content. Carefully controlled extraction and efficient preliminary chromatographic separations were set up. Deuterium labelled analogues were used as internal standards for the quantitative determination by gas chromatography-mass spectrometry and some new derivatives were included in the identification procedure. The identified estrogens and their mean concentrations (ng/ml) are: 2-hydroxy-estrone (0.14), 4-hydroxy-estrone (0.12), 2-hydroxy-estradiol (0.36), 4-hydroxy-estradiol (0.34), 6 alpha-hydroxy-estradiol (13.2), 6 beta-hydroxy-estradiol (6.40), 2-methoxy-estrone (0.83), 2-methoxy-estradiol (10.5), 16-oxo-estradiol (0.41), estriol (10.2), estradiol-17 beta (1365), estradiol-17 alpha (1.91), estrone (211). Metabolism of estradiol by 6-hydroxylation seems to be predominant in the human ovary. The other data suggest that 2-hydroxylation, with subsequent O-methylation, and 16-hydroxylation may be by equivalent pathways, since the sum of the 2-methoxy-estrone and 2-methoxy estradiol concentrations is rather similar to the estriol concentration. Hence, the latter three compounds and the 6-hydroxy-estradiols may be end-products of follicular estrogen metabolism. Catechol estrogen formation by 2-hydroxylation and 4-hydroxylation is shown to be of equal importance in the ovary. These results confirm the presence in the human follicle of various competing estrogen hydroxylases and catechol-O-methyltransferase.

Estrogens↗