EFFECT OF TOXIC URINARY EXTRACTS ON SPECIFIC ASSAYS FOR HUMAN CHORIONIC GONADOTROPHIN AND FOLLICLE-STIMULATING HORMONE

1970 ◽  
Vol 65 (3) ◽  
pp. 552-564
Author(s):  
L. J. Hipkin

ABSTRACT Boiled urinary extracts, which had inhibited the activity of human chorionic gonadotrophin (HCG) in the mouse uterus assay, potentiated the effects of HCG and follicle-stimulating hormone (FSH) in the ovarian ascorbic acid depletion and rat ovarian augmentation assays respectively. This treatment inhibited responses to FSH when the ovarian augmentation assay was conducted in hypophysectomised animals. A toxic urinary extract did not affect the activity of HCG in the ventral prostate weight assay in either intact or hypophysectomised rats. Reasons for the different effects of boiled urinary extracts in these assays are discussed.

1962 ◽  
Vol 24 (2) ◽  
pp. 153-158 ◽  
Author(s):  
H. SCHMIDT-ELMENDORFF ◽  
J. A. LORAINE ◽  
E. T. BELL

SUMMARY The luteinizing hormone (LH), follicle-stimulating hormone (FSH) and 'total gonadotrophic' activities of various hormones have been studied following incubation with 6 m urea at 40° c for 24 hr. LH activity was estimated by the ovarian ascorbic acid depletion test in rats, FSH activity by the augmentation test in mice, and 'total gonadotrophic' activity by the mouse uterus test. Following incubation with 6 m urea the LH activities of NIH—LH, NIH—FSH, human chorionic gonadotrophin and Pergonal were almost completely destroyed, while the LH activity of pregnant mares' serum gonadotrophin (PMSG) was reduced to a smaller extent. The FSH activity of NIH—FSH was little affected by this form of treatment, but in the case of Pergonal a considerable reduction of FSH activity occurred. The 'total gonadotrophic' activity of NIH—FSH, PMSG and Pergonal was reduced after incubation with 6 m urea, the degree of inactivation being greatest in the case of Pergonal. After control incubations with water no appreciable loss of biological activity was observed with any hormone other than Pergonal.


1969 ◽  
Vol 7 (9) ◽  
pp. 33-35

The three substances now used to stimulate the gonads in infertility are human follicle stimulating hormone (HFSH) obtained mainly from post-menopausal urine, but also from human pituitary glands, human chorionic gonadotrophin (HCG) extracted from the urine of pregnant women, and clomiphene (Clomid - Merrell), a synthetic compound which we reviewed in 1967.1


1978 ◽  
Vol 89 (1) ◽  
pp. 166-172 ◽  
Author(s):  
T. J. Weiss ◽  
D. T. Armstrong ◽  
J. E. A. McIntosh ◽  
R. F. Seamark

ABSTRACT Theca and granulosa tissues isolated from sheep ovarian follicles of different sizes were incubated in the presence of human chorionic gonadotrophin (HCG; 5 IU/ml) or follicle stimulating hormone (FSH; 5 μg NIH-FSH-S11/ml) for 40 min. Changes in the total amounts of cyclic 3′,5′-adenosine monophosphate (cAMP) were used as an index of the responsiveness of these preparations to the hormones. Thecal tissue of both large (4–6 mm in diameter) and small (1–3 mm) follicles responded similarly to gonadotrophins. Granulosa cells from small follicles failed to respond to stimulation by HCG. FSH, however, consistently increased cAMP production in comparison with controls or cells treated with HCG. Granulosa cells of large follicles responded to both HCG and FSH.


1971 ◽  
Vol 67 (2) ◽  
pp. 262-276 ◽  
Author(s):  
P. Petrusz ◽  
C. Robyn ◽  
E. Diczfalusy

ABSTRACT Forty-two antisera were prepared in rabbits against human chorionic gonadotrophin (HCG), human hypophysial gonadotrophin (HHG), human urinary luteinizing hormone (LH) and human menopausal gonadotrophin (HMG) preparations. The gonadotrophic profiles of the antigens were previously characterized by bioassay, immunoassay and bioimmunoassay methods. The 25 most potent antisera were tested in statistically valid bioassays for their HCG and follicle stimulating hormone (FSH) neutralizing activities as well as for their neutralizing potencies against the FSH-like activity present in HCG preparations. The anti-HCG/anti-FSH ratios of the anti-HCG sera tested varied between 6.2 and > 254, while those of the anti-HHG, anti-LH and anti-HMG sera were close to 2. It was found that the total dose of immunological activity (anti-HCG neutralizing and anti-FSH neutralizing potency) rather than that of the biological activity administered to the rabbits was decisive for obtaining antisera with high anti-HCG and anti-FSH titers. Immunization with a highly purified HCG preparation (> 17 000 IU/mg) resulted in antisera exhibiting lower anti-HCG/anti-FSH ratios than did immunization with partially purified preparations. A highly purified urinary LH preparation which did not contain any detectable FSH activity gave rise to antisera exhibiting anti-HCG/anti-FSH ratios of approximately 2.0. These highly purified HCG and LH preparations were shown previously to possess high anti-FSH neutralizing potencies (Petrusz et al. 1971b). Booster injections did not change significantly the quality or the titer of the antigonadotrophic sera studied. The HCG neutralizing potency of anti-HCG sera was approximately 3 times higher when assayed against a highly purified HCG preparation (> 17 000 IU/mg) as compared to potency estimates obtained against the laboratory standard of HCG (about 2000 IU/mg). It is suggested that consideration should be given to the establishment of standard preparations of antigonadotrophic sera. It is concluded that bioimmunoassays are more suitably than conventional bioassay methods for the assessment of the antigenic purity of human gonadotrophin preparations.


2007 ◽  
Vol 19 (1) ◽  
pp. 288
Author(s):  
M.-K. Kim ◽  
H.-J. Oh ◽  
Y. H. Fibrianto ◽  
G. Jang ◽  
H.-J. Kim ◽  
...  

Growth, maturation, and ovulation of the Graafian follicle depend on appropriate patterns of secretion, sufficient concentrations, and adequate ratios of various reproductive hormones, especially follicle stimulating hormone (FSH) and luteinizing hormone (LH) (Van Tol et al. 1996 Mol. Reprod. Dev. 45, 218–224). The present study investigated the effects of FSH and human chorionic gonadotrophin (hCG) on the nuclear maturation of canine oocytes. In addition, in order to investigate the effect of stage of the estrous cycle on the meiotic competence of canine oocytes matured in vitro, oocytes were collected from various reproductive states and matured in vitro in the presence of the gonadotrophins. Estrous cycle stage was evaluated for each bitch by ovarian morphology, and bitches were categorized according to the stage of the estrous cycle (anestrus, follicular, or diestrus) prior to oocyte collection. Recovered oocytes were cultured in serum-free tissue culture medium (TCM)-199 supplemented with various concentrations of FSH (Exp. 1: 0, 0.1, 1.0, or 10 IU) or hCG (Exp. 2: 0, 0.5, 1.0, or 10 IU) or both (Exp. 3: 1 IU FSH + 1 IU hCG) for 72 h to determine the effective concentration of these hormones, and to examine their combined effect. After maturation culture, oocytes were denuded in PBS containing 0.1% (w/v) hyaluronidase by gentle pipetting. The denuded oocytes were stained with Hoechst 33342 in glycerol and the nuclear state of oocytes was evaluated under UV light. The rates of maturation to the MII stage were significantly higher (P < 0.05) when follicular-stage oocytes were supplemented with 1 IU FSH (6.2%) compared with the other FSH-supplemented groups (0.0 to 3.3%) or to the control (1.8%), or 0.1 or 10.0 IU FSH (0 to 1.2%). Significantly higher (P < 0.05) maturation rate to MII stage was observed in follicular-stage oocytes supplemented with 1.0 IU hCG (7.2%) compared with the control or other hCG-supplemented groups (0 to 1.5%). However, FSH and hCG together did not improve the nuclear maturation rate of canine oocytes (2.4%) compared with FSH (6.2%) and hCG alone (7.2%). In conclusion, FSH or hCG alone significantly increased the maturation of canine oocytes to the MII stage. This work was supported by grant No. M1062503005-06N250300510 from KOSEF, Republic of Korea.


1966 ◽  
Vol 35 (2) ◽  
pp. 199-206 ◽  
Author(s):  
P. S. BROWN ◽  
M. WELLS

SUMMARY The follicle-stimulating hormone (FSH) content of urinary gonadotrophic extracts was assayed by its effect on the ovarian weight of immature mice when given in conjunction with 40 i.u. human chorionic gonadotrophin. About three-quarters of all routine assays gave values of λ between 0·15 and 0·30. Precision was slightly increased when the material was given in three rather than in five injections. Correction of ovarian weight for body weight was either invalid or of no value in reducing variance. Removal of between-litter variance increased precision considerably. Mice of three randomly bred colonies were all satisfactory, and inbred C57BL mice were also suitable for the assay. C3H mice were less sensitive. The efficiency of different methods of extracting FSH from urine was examined. The method of Johnsen (1958) using precipitation with tannic acid was considered the most satisfactory and gave extracts of high potency and low bulk. Limited experiments in which purified human pituitary FSH was assayed with and without added luteinizing hormone, gave results compatible with the assumption that the method is specific for FSH.


1976 ◽  
Vol 70 (3) ◽  
pp. 335-344 ◽  
Author(s):  
J. I. THORELL ◽  
B. HOLMSTRÖM

SUMMARY Antisera were produced in rabbits against highly purified preparations of human LH (2000 or 10000 i.u./mg), human FSH (5500 i.u./mg), and human TSH (7·5 i.u./mg). Most rabbits produced antisera of high titre and high avidity. Cross-reactions were minimal between human TSH and human chorionic gonadotrophin (HCG) and between human FSH and HCG but marked between human LH and HCG. TSH and FSH also showed a constant but relatively weak cross-reaction. LH cross-reacted with FSH to a higher degree than did HCG. The avidity of the antisera was high. It was concluded that much of the lack of specificity recorded for glycoprotein antisera are effects of impure immunogens. Some of the true cross-reactions are probably explained by shared antigenic determinants of the β-subunits. Unadsorbed antisera could be used for assay of FSH and TSH in plasma from pregnant women.


1978 ◽  
Vol 79 (3) ◽  
pp. 407-408 ◽  
Author(s):  
M. J. ELLIS ◽  
R. A. DONALD ◽  
J. H. LIVESEY

The Medical Unit, The Princess Margaret Hospital, Christchurch 2, New Zealand (Revised manuscript received 21 August 1978) The frequent clinical and research requirement for measurement of both plasma luteinizing hormone (LH) and follicle-stimulating hormone (FSH) has prompted the development of a simultaneous radioimmunoassay for these two hormones. The considerable advantages of a simultaneous method include an economy of plasma volume, assay reagents, test-tubes and, more importantly, the time required for radioactive counting and performance of the assay by technical staff. The latter two factors comprise a significant proportion of radioimmunoassay operating costs. This report describes a simultaneous radioimmunoassay based on the use of 131I-labelled FSH, 125I-labelled LH, anti-FSH serum M93 6873 (a generous gift from Professor W. R. Butt, Birmingham), anti-human chorionic gonadotrophin (HCG) serum for LH measurement (Donald, 1972) and donkey anti-rabbit precipitating serum (Wellcome Reagents, U.K.) for separation of antibody-bound and free hormones. Pituitary gonadotrophin standard (LER 907)


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