Human Pituitary Gonadotropin Research Articles

The use of human ovarian responsiveness in a new bioassay for follicle-stimulating hormone.

ABSTRACT In a program of ovulation induction, 88 patients received more than one course of treatment with the same batch of human pituitary gonadotropin (hPG); 81 of these patients responded reproducibly in their repeat cycles. Of these, 68 also received at least two different batches of hPG in a total of 225 satisfactory treatment cycles. A model was developed which allowed the calculation of the relative patient potencies from the data obtained and the results were compared with the FSH and LH contents of the batches estimated by radioimmunoassay and with FSH estimates by the Steelman-Pohley and P. S. Brown bioassays. The results of the human assay correlated well with FSH values obtained by radioimmunoassay, less well with the FSH values obtained by the P. S. Brown assay, poorly with the values obtained by the Steelman-Pohley assay and not at all with the LH values obtained by radioimmunoassay after correction for the FSH effect. One batch which did not show this correlation had an exceptionally short ...

Hypogonadotrophic Hypogonadism Associated with Anosmia in the Female

Summary: Three women with hypogonadotrophic hypogonadism associated with anosmia were subjected to ovarian stimulation with human gonadotrophins. The first responded normally to conventional dosage of human pituitary gonadotrophins (HPG). The second, known to have primordial follicles in the ovary, failed to respond to massive doses of HPG. The third, who was unmarried, failed to respond to an intramuscular injection of 75 I.U. of human menopausal gonadotrophs daily for 3 days. The importance of testing olfactory acuity in patients with menstrual irregularities is re‐emphasised.

Induction of ovulation in patients following removal of a pituitary adenoma

Five infertile women who were operated upon for pituitary adenomas were treated with human pituitary, postmenopausal, and human chorionic gonadotropins in order to conceive. Ovulation took place following each treatment. Four of the women conceived; three gave birth to twins and one to a single infant. One of the women went into labor spontaneously; three, close to term, were delivered by cesarean section. The seven children were normal and healthy.

Gonadotropins in the infant chimpanzee: a sex difference.

SummarySerial circulating levels of FSH were higher in five female than in four male infant (0.4-2.0 years) chimpanzees. A rhythm in FSH levels was observed in the females with a periodicity of 8–25 days. There was no sex difference in mean LH levels. However, greater day-to-day variability was observed in the females.The animals used in the study were handled in accordance with the “Guide for Laboratory-Animal Facilities and Care” established by The National Academy of Science-National Research Council. This work was supported in part by M. R. C. Canada Grant MA-2997 and by N. I. H. Contract DRR-73-2100. The LER-907 human pituitary gonadotropin standard was a gift of the National Pituitary Agency. We thank Dr. F. S. Chebib for help with the statistical analyses and Mrs. R. Poturnak, Mrs. E. Dudeck, and Mr. D. Bookout for technical assistance.

Use of l-dopa (l-dihydroxyphenylalanine) to stimulate ovulation

This study was prompted by previous experimental data which suggested that l-dopa may be useful clinically in treating anovulation and oligoamenorrhea due to hypothalamic-pituitary hypofunction of nonorganic origin. Six such patients, who had failed to respond to clomiphene therapy but who had previously ovulated and become pregnant following the administration of human pituitary gonadotropins, were treated with 2 Gm. of l-dopa daily for 3 months. There was no response in 3 patients. The remaining 3 patients showed evidence of increased gonadotropin and estrogen activity. However, follicular maturation did not reach the level necessary to trigger ovulation, even with the addition of human chorionic gonadotropin. These data indicated that in the manner used in this study l-dopa is not useful clinically in hypogonadotropism.

COMBINED INTRA-UTERINE AND EXTRA-UTERINE PREGNANCY IN TWO PATIENTS TREATED WITH HUMAN PITUITARY GONADOTROPHINS

Summary: Two cases of combined intra-uterine and extra-uterine pregnancy in patients receiving treatment with human pituitary gonadotrophin for stimulation of ovulation are described. One patient subsequently delivered a healthy child, whereas the other miscarried before diagnosis of the extra-uterine pregnancy was made.

Species Differences in Luteinizing Hormone as Inferred from Slope Variations in a Radioligand Receptor Assay 1

Luteinizing hormone preparations from a variety of sources were compared with respect to the slopes of their radioligand uptake inhibition curves. The system utilized human luteinizing hormone as the radioligand and homogenates of testis from mature rats as the receptor source. Based on the weighted mean values from replicate slope measurements, four major groups could be detected. In order of decreasing slope steepness, these were: Group I = human urinary LH, monkey pituitary LH, human pituitary LH and human chorionic gonadotropin; Group II = bovine pituitary LH, ovine pituitary LH, rat pituitary LH and equine pituitary LH; Group III = rabbit pituitary LH and chicken pituitary LH; Group IV = porcine pituitary LH and canine pituitary LH. There was no overlap of 95% fiducial limits for calculated slope values between groups. These groupings are at variance in some instances with those of a previous report (Endocrinology 73: 509, 1963), based on slopes of dose response curves obtained using the ventral pros...

Regulation of the Corpus Luteum During the Menstrual Cycle and Early Pregnancy

Regulation of the corpus luteum during the menstrual cycle and early pregnancy is reviewed. Data relevant to influential factors such as gonadotropic hormones from the pituitary the products of conception the uterus and the local influence of steroid hormones are discussed. Data described suggest that the human corpus luteum may not need continued support by gonadotropic hormone however these data were difficult to interpret because no human pituitary gonadotropins were available for studies and because endogeneous gonadotropin levels were not determined prior to and during treatment. A summary of the data suggests that luteinzing hormone (and/or human chorionic gonadotropin in pregnancy) is the prinicipal luteotropic factor in women. Mechanisms of action are discussed.

Antigenic structure of human gonadotrophins: pituitary luteinizing hormone.

SUMMARY Immunological relationship between human pituitary luteinizing hormone (LH) and human chorionic gonadotrophin (HCG) was measured by microcomplement fixation, and a value of 1·17 was obtained for the immunological index of dissimilarity determined from the reactions of a highly purified human LH with rabbit antiserum to HCG. Treatment with neuraminidase of the LH preparation did not affect either the biological activity, as measured by ovarian ascorbic acid depletion, or the complement-fixing activity. The biological and immunological activities of LH were also not influenced by treatment with β-glucosidase or with neuraminidase plus β-glucosidase. Partial tryptic hydrolysis of LH caused decreases in both the biological and immunological activities, the greater loss being in the former. The biological and immunological activities of LH were nearly completely destroyed by treatment with urea, and removal of urea by dialysis did not restore the two activities. When the LH preparation was exposed to increased temperatures for various periods of time or to extremes of pH, the two activities were affected to variable degrees. These results indicate that the protein but not the carbohydrate moiety of the LH molecule is essential for both biological and immunological activities of the hormone and that the conformational integrity of certain parts of the protein moiety is also important for these activities.

Treatment of primary amenorrhoea with human pituitary and chorionic gonadotrophins.

SummaryFifteen infertile women with primary amenorrhoea and pronounced oestrogen deficiency were treated with human pituitary gonadotrophin and human chorionic gonadotrophin 71 times; 61 ovulations (86 per cent) were recorded. Eleven patients conceived (73·5 per cent), three of them twice and one three times, giving 16 pregnancies in all. There were 12 single births and four sets of twins. Two pregnancies ended in abortion. On average one pregnancy resulted from 4·5 treatments and clinical symptoms of overstimulation occurred in 2 cycles (2·8 per cent).

Recent studies on the estimation of human pituitary gonadotrophins in urine in clinical conditions.

(a) In women approaching the menopause urinary LHlevels tend to be high while those of FSH are within the normal range for subjects during the reproductive life. In a proportion of women in this group high LH levels can occur in absence of any clinical symptoms of the menopause, (b) In postmenopausal subjects urinary LH output is significantly higher than in normally menstruating women and is in the same range as that found in the perimenopausal group, (c) In patients with Turner’s syndrome excretion values for LH are variable and are not correlated with the chromosomal karyotype. In phenotypic males with a 47, XYY sex chromosomal constitution, urinary LH readings are grossly elevated, (d) In infertile women treated with clomiphene a significant effect on urinary LH output was not observed, suggesting that the compound did not act directly on the pituitary-hypothalamic axis to produce release of this hormone, (e) In patients treated on a long term basis with oral contraceptives, urinary LH activity was not completely suppressed, levels being within or at the lower end of the normal range for women during reproductive life.

Sialic acid content and sperm receptivity of cervical mucus in relation to oestrogen excretion following administration of FSH.

ABSTRACT The cervical mucus was studied with regard to changes in sialic acid content and sperm receptivity. The data were correlated with the urinary total oestrogen excretion in anovulatory women treated with human pituitary gonadotrophins (HPG). With increasing total oestrogen excretion the sialic acid content decreased and the sperm receptivity increased. At a total oestrogen level of about 60 μg/24 h the respective patterns of the curves were reversed and higher values for total oestrogen were actually associated with a decrease in sperm receptivity and an increase in sialic acid content. The possible significance of these observations is discussed.

Pituitary gonadotropin assays.

The methodology of gonadotropin assays has moved steadily during its 35 year evolution toward specificity and greater sensitivity. The three phases of this development are represented by the bioassay of total human pituitary gonadotropins in urine, specific bioassays for follicle-stimulating and luteinizing hormones in urine, and the radioimmunoassay for these hormones in serum or plasma.

The relation between plasma progesterone and total urinary oestrogens following induction of ovulation in women.

ABSTRACT Plasma progesterone levels have been measured before and after a single injection of 9000 IU of human chorionic gonadotrophin (HCG)* at different levels of total oestrogen excretion (TE) in 67 treatments with human pituitary gonadotrophin (HPG) in amenorrhoic women. With a TE excretion of less than 70 μg/24 h the rise in plasma progesterone was similar to that found in a normal menstrual cycle indicating the formation of only one corpus luteum. With more than 70 μg/24 h of TE several corpus lutea was formed as judged by the rise in plasma progesterone levels. No further increase was seen above 300 μg/24 h. The luteal phase was shorter than normal in the group with 75–300 μg/24 h of TE.

A quantitative immunological assay for pregnant mare serum gonadotrophin.

SUMMARY Rabbits and turkeys were immunized against commercial preparations of pregnant mare serum gonadotrophin (PMSG). Ouchterlony double diffusion and haemagglutination-titration tests showed that the rabbit antisera contained non-specific antibodies against horse serum proteins, whereas the turkey antisera did not. Both types of antisera neutralized the effects of PMSG in vivo. One specific turkey anti-PMSG serum was used unabsorbed to establish a sensitive and accurate haemagglutination—inhibition assay for PMSG in equine serum and in saline. Pyruvic aldehyde-treated sheep red blood cells sensitized with PMSG were used in perspex haemagglutination trays. The reliability criteria of the assay is discussed. The smallest quantity of PMSG detectable in saline was 0·3 i.u./ml. The turkey anti-PMSG serum showed a strong immunological cross-reaction with preparations of equine pituitary follicle-stimulating hormone (FSH) and equine pituitary luteinizing hormone (LH), a faint reaction with human pituitary gonadotrophin and no reaction against human chorionic gonadotrophin, ovine FSH, ovine LH, bovine LH or porcine FSH.

Differential effect of chymotryptic digestion on the biological activity of pituitary, serum and urinary gonadotropins.

1. The luteinizing hormone activity in gonadotropin preparations from human and equine pituitaries, human chorionic gonadotropin and pregnant mare serum was inactivated by digestion with alpha-chymotrypsin. By contrast, the luteinizing hormone activity of a human postmenopausal urinary gonadotropin preparation was resistant to enzymic digestion. 2. As with human, ovine and porcine pituitary gonadotropin preparations, human postmenopausal urinary follicle stimulating hormone was not inactivatedinactivated by chymotryptic digestion. This hormonal activity, however, was destroyed by the enzyme in preparations from equine pituitaries and pregnant mare serum. 3. The results suggest that differences exist between pituitary and urinary LH, and between equine FSH and FSH from the other sources studied. These dissimilarities are presumably a reflection of structural differences in the neighborhood of the biologically active site on these hormones. (Endocrinology 84: 9, 1969)

BIOASSAY OF ANTIGONADOTROPHIC SERA

ABSTRACT Methods are described for the bioassay of the human follicle stimulating hormone (FSH) neutralising potency of antigonadotrophic sera. The methods are based on a modified ovarian weight augmentation test using human chorionic gonadotrophin (HCG) or luteinising hormone (LH) of ovine origin for augmentation. The antigonadotrophic sera were obtained following immunisation of rabbits with HCG, human menopausal gonadotrophin (HMG) and human hypophysial gonadotrophin (HHG) preparations. The FSH neutralising potencies of these antisera were assayed against laboratory standard preparations of HMG and HHG and against the Second International Reference Preparation of HMG. When HCG was used for augmentation, the FSH neutralising potency of antisera depended on the sequence in which HCG, HMG and antiserum were combined. When HCG was mixed with the antiserum prior to the addition of HMG, this resulted in a significant decrease in the FSH neutralising potency. When HCG was injected separately from the HMG-antiserum complex, the FSH neutralising potency increased. However, the FSH neutralising potency of all antisera was significantly higher when LH of ovine origin, rather than HCG was used for augmentation. Anti-HCG sera exhibited a considerable FSH neutralising potency, even when prepared by immunisation with HCG preparations of high specific activity. These high FSH neutralising potencies were in contrast to the low FSH activity of the HCG preparations used for immunisation. Anti-HMG sera possessed little, if any, FSH neutralising potency. These poor FSH neutralising potencies were in contrast to the high FSH activity of the HMG preparations used for immunisation. The FSH neutralising potency of an anti-HHG serum was at least 5 times higher when assayed against HMG, than when assayed against HHG. The data presented indicate that HCG preparations extensively compete with FSH preparations for antibodies neutralising FSH activity. This suggests that there is a cross reaction between HCG and FSH. The data also indicate, that there are significant differences in the antigenic properties of human pituitary and urinary gonadotrophins. It is concluded, that the establishment of specificity of immunoassay methods for human gonadotrophins cannot be based exclusively on immunological evidence. Also, the absorption procedures used to improve the specificity of antigens and antisera are of limited value, unless carried out in a strictly quantitative manner following the establishment of the profile of the gonadotrophic and antigonadotrophic activities present.

Clinical Evaluation of Ethinylestrenol and Mestranol as a Contraceptive

Ethinylestrenol (2.5 mg) plus mestranol (75 mcg) was clinically evaluated as an oral contraceptive (trade name Lyndiol) by cyclically administering the combination to 625 women for 9898 cycles (1-31 cycles each). Blood tests were performed in 2 groups of 25 patients each at 0 3 6 12 and 18 months to determine if there was any effect on hemoglobin hematocrit red cell white cell or platelet counts fibrinogen recalcified plasma clotting time clotting time quick prothrombin time plasminogen partial thromboplastin generation bromsulphonphthalein retention total protein bilirubin alkaline phosphatase serum glutamic pyruvic transaminase LE cell preparation cephalin flocculation urea nitrogen calcium phosphorus and protein-bound iodine. In 25 other patients 24-hour urine specimens were examined initially at 3 months and at 1 year for pregnanediol 17-ketosteroids 11-oxysteroids and human pituitary gonadotropins. The only significant difference was a reduction in production of pregnanediol during therapy. 323 patients (52%) dropped out before study termination: 181 moved and were not located 50 (8%) discontinued of their own accord due to side effects and 22 (4%) dropped out on advice of physician. Side effects per cycle encountered most frequently were intermenstrual spotting (in 2.6% of patients) headache (2.2% of patients) and nausea and gastric distress (1.6%). 4 pregnancies occurred in women taking the pills incorrectly. 1 patient had prolonged amenorrhea until 6 months after contraceptive discontinuation when menses resumed spontaneously. Lyndiol is considered an effective contraceptive combination.

Immunologic reaction between human luteinizing hormones and antiserum to urinary luteinizing hormone.

Antiserum was produced in rabbits to a partially purified LH fraction of human postmenopausal gonadotropin (HMG) and then absorbed with human serum and a potent FSH preparation from HMG. The absorbed antiserum appeared to be serologically specific to human urinary LH. This antiserum gave no demonstrable precipitin reaction with either a highly purified human pituitary LH preparation or serum gonadotropin, but did inhibit the OAAD activities of these 2 preparations. Neither precipitation nor neutralization occurred between the antiserum and HCG. The degree of immunologic cross-reactivity among various human LH preparations was examined by quantitative micro-complement fixation with the absorbed antiserum, and the following values of the immunologic index of dissimilarity were obtained: urinary LH 1.00, serum gonadotropin 3.1, pituitary LH 6.0 and HCG 1.0. Complement-fixing activities of nonpregnant urinary LH preparations were in good agreement with their LH potencies; however, this did not hold true for H...

Gonadotrophin stimulation test ovarian function.

Dynamic tests of endocrine function have proved their value in the clinical study of the thyroid and adrenal glands, but their development for the assessment of gonadal function has been inconspicuous. Now that effective means of treating some disorders of ovarian function are available?even if at present only to a limited extent?the reliable determination of ovarian sensitivity to gonadotrophic stimulation has become a matter of practical clinical importance. This is particularly true when treatment with human gonadotrophic preparations is contem plated, not only in order to avoid the wastage of scarce material in cases which could not be expected to respond but also to minimize the real dangers of overstimulation of unduly sensitive ovaries (Mozes et al., 1965). As has been shown by Crooke et al. (1966) the dosage of human follicle-stimulating hormone (F.S.H.) required to produce a positive ovarian response varies in different women over at least an eightfold range. In woman with amenorrhoea it has long been customary to differentiate primary and secondary ovarian failure on the basis of the urinary gonadotrophin excretion, the mouse uterine weight response ( total gonadotrophin ) being the basis of the test most widely used. An increased excretion is interpreted as an indication of intrinsic ovarian failure, the absence of the negative feed-back effect of ovarian oestrogen being the cause of excessive F.S.H. secretion by the pituitary gland ; a low excretion is held to imply secondary failure, the ovary being inadequately stimulated by the deficient pituitary gonadotrophin production. Apart from the inherent defects of this bioassay procedure, which introduce a substantial degree of unreliability as well as inconvenience for general clinical use, determination of urinary gonadotrophin excretion fails to give any idea of whether ovaries which can respond to gonadotrophic stimula tion are sensitive to such stimulation in greater or less degree. We doubt, indeed, whether increased urinary total gonadotro phin necessarily implies ovarian unresponsiveness, and we have good evidence that ovarian unresponsiveness may be found in some patients without increased urinary gonadotrophin excretion, as measured by the mouse uterine weight assay. It was awareness of these needs and drawbacks which persuaded Shearman (1964) to propose the determination of the response of the urinary excretion of oestrone to injections of pregnant mares' serum gonadotrophin (P.M.S.) as a test of ovarian responsiveness to gonadotrophic stimulation in the human female. His brief report showed that in the 18 patients he studied intramuscular injections of 5,000 i.u. daily for three days produced significant rises in urinary oestrone excretion in 13 women with normal ovaries, no response in one patient with ovarian dysgenesis, and elevated responses in four women with polycystic ovaries. These findings suggest that human ovaries respond to P.M.S. in much the same way as they do to human pituitary or menopausal gonadotrophin. Because P.M.S. preparations are marketed, are readily available, and are much cheaper than human gonadotrophic material, their use for testing ovarian responsiveness would clearly be advantageous. The procedure as carried out by Shearman (1964) involved the daily collection of 24-hour urine specimens for eight con secutive days, the first two providing a baseline, and the administration of three daily injections of gonadotrophin. This seemed to us to be too involved for routine clinical use on a wide scale. Moreover, it was not clear whether the peak of oestrone excretion had been reached within the time covered by Shearman's observations, and we therefore undertook a study with a view to evolving, if possible, a simpler procedure, the ultimate objec tive requiring no more than one 24 hour urine collection and a single gonadotrophin injection. In this we believe we have been successful.