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Search for other papers by J. A. LORAINE in
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SUMMARY
Pooled urine from normal men, normally menstruating women and hospitalized postmenopausal subjects was extracted by six published methods and assayed for human pituitary gonadotrophic (HPG) activity by the mouse uterus test in terms of HMG-20A.
In the three types of urine studied comparable yields of HPG were obtained by all six extraction methods, and an analysis of variance showed the absence of significant differences between the methods.
No evidence could be obtained to support previous claims that the kaolinacetone method of Loraine & Brown (1959) gave less satisfactory yields of urinary HPG than certain of the other procedures.
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A number of methods have been proposed for the biological assay of human chorionic gonadotrophin (c.g.). The test animals have usually been immature or hypophysectomized rats or mice in which circulating gonadotrophins were lacking.
The index of response has varied greatly [Thayer, 1946]. Tests have been based on observations of corpora lutea [Aschheim & Zondek, 1927; Emmens, 1939; Kennedy, 1933; Siegler, 1939; Smith & Smith, 1934, 1935, 1939, 1948; White, 1937; White & Hunt, 1943]; ovarian weight [Evans, Kohls & Wonder, 1937; Evans, Meyer & Simpson, 1929, 1931, 1933; Wallen-Lawrence & Van Dyke, 1931]; uterine weight [Delfs, 1941, Emmens, 1939; Dorfman, Rubin & Miller, 1946, 1947; Jones, Delfs & Stran, 1944; Sealey & Sondern, 1940]; vaginal smears [Browne & Venning, 1936a, b, 1938; Venning, 1948; Venning & Browne, 1936; Wallen-Lawrence & Van Dyke, 1931]; vaginal smears in vitamin B-deficient rats [Heard, 1941; Heard & Winton, 1939]; seminal vesicles [Bischoff,
Search for other papers by J. A. LORAINE in
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Search for other papers by J. B. BROWN in
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SUMMARY
A method for the quantitative determination of gonadotrophins in the urine of non-pregnant human subjects is described.
The reliability criteria for the method have been studied and the technique has been found to be satisfactory in terms of accuracy, precision, sensitivity and specificity.
The importance of expressing results of urinary gonadotrophin assays in terms of a reference preparation such as HMG 20A is emphasized.
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Search for other papers by J. A. LORAINE in
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SUMMARY
The assay method for luteinizing hormone (LH) activity depending on ovarian ascorbic acid depletion in rats (O.A.A.D. method) has been assessed in terms of its reliability criteria.
The chief advantages of the method are its high degree of sensitivity and specificity and its good practicability. Its disadvantages are its relatively low degree of precision and the fact that a proportion of assays are invalid due to lack of parallelism.
The LH activity of various gonadotrophin preparations has been investigated using the O.A.A.D. method. The LH content of NIH-FSH and of Pergonal was found to be low. The LH activity of pregnant mare's serum gonadotrophin was lower than that of human chorionic gonadotrophin.
When purified urinary extracts are used, the O.A.A.D. method is suitable for clinical application.
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Search for other papers by E. DICZFALUSY in
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SUMMARY
The influence of prolactin on the assay of human menopausal gonadotrophin (HMG) and human chorionic gonadotrophin (HCG) has been studied. Various bioassay methods were used and the experiments were conducted in two independent laboratories.
In hypophysectomized immature male rats the response of the ventral lobe of the prostate to HMG was not affected by simultaneous administration of prolactin. It was concluded that, in the case of urinary extracts, prolactin did not interfere with the specificity of this test for interstitial cell stimulating hormone activity.
Valid assays of HMG by the rat uterus and mouse uterus tests could be obtained in the presence of relatively large quantities of prolactin.
When HCG was assayed by the rat uterus test and by tests depending on the enlargement of the accessory reproductive organs in male rats, simultaneous administration of prolactin did not affect the results obtained.
In immature male rats prolactin did not prevent the regression of the accessory organs which occurs after castration.
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Search for other papers by J. A. LORAINE in
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SUMMARY
1. The urinary excretion of oestrogens, pregnanediol and pituitary gonadotrophins has been studied throughout nine ovulatory menstrual cycles.
2. The pattern of hormone excretion was relatively constant from one individual to another, but the actual amounts excreted varied considerably in different individuals.
3. In none of the subjects studied did the mid-cycle peak in gonadotrophin precede the oestrogen peak.
4. The increase in urinary pregnanediol during the luteal phase occurred at the same time as or just before the rise in basal temperature and 1–4 days after the oestrogen peak.
5. There was no correlation between the amounts of oestrogens and pregnanediol excreted during the luteal phase of the cycle.
6. When gonadotrophin assays were conducted by the mouse uterus test and that depending on the prostate of the hypophysectomized rat, the results obtained agreed very closely at all stages of the cycle.
7. In one subject a marked rise in gonadotrophin output was observed as early as 9 days after a successful artificial insemination.
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Search for other papers by H. A. ROBERTSON in
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1. The ACTH concentration has been estimated in lyophilized pituitary tissue from five adults, seven infants and eight foetuses. Results are expressed in terms of the international standard. Adult pituitaries contain more ACTH than infantile glands, and infantile glands more than foetal ones.
2. ACTH activity was detected in foetal pituitaries at a period of gestation as early as 16 weeks.
3. The accuracy of the Sayers test was considerably improved when the time interval between hypophysectomy and assay was increased from 24 to 48 hr.
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Search for other papers by HELEN E. C. CARGILL THOMPSON in
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Search for other papers by J. A. LORAINE in
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SUMMARY
1. Unilateral section of the sciatic nerve causes atrophy of the gastrocnemius muscle with consequent reduction in wet weight and in fibre size of the muscle in both hypophysectomized and non-hypophysectomized animals.
2. The administration of growth hormone, thyroid-stimulating hormone (TSH), follicle-stimulating, luteinizing and adrenocorticotrophic hormone, either before nerve section, immediately after nerve section or some time after atrophy has been established, had no effect on muscle fibre size.
3. The administration of TSH during the period immediately following nerve section caused a significant reduction in the tibial epiphysial cartilage width of the denervated limb. No difference in cartilage width between denervated and innervated limbs was observed following treatment with any of the other hormones.
4. In both normal and hypophysectomized animals the effect of the hormones on body weight and on the weight of normal and atrophic muscles was in keeping with their known physiological actions.
5. It is concluded that anterior pituitary hormones act on muscles by virtue of their well-known anabolic or catabolic properties, that their effects on total body weight and innervated and denervated muscle are generally similar and that, under the experimental conditions used, none of the hormones studied was capable of preventing the occurrence of atrophy in denervated muscle.
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Search for other papers by J. A. LORAINE in
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SUMMARY
1. The effect of the progestational compounds, norethisterone and norethisterone acetate, on ovarian and pituitary function during the menstrual cycle have been assessed in five subjects by measuring the urinary excretion of oestrogens, pregnanediol, pregnanetriol and pituitary gonadotrophin (HPG).
2. The effects of the two compounds were similar and depended on dosage and time of administration during the cycle. With adequate dosages given from day 5 to day 25, there was complete suppression of ovarian activity; with lower dosages given over a similar period, follicular activity could persist but ovulation and formation of a corpus luteum were suppressed. When administered during the luteal phase only, there was no apparent effect on the normal function of the corpus luteum. In none of the subjects was urinary HPG excretion depressed during drug administration.
3. The action of these compounds as ovarian or pituitary inhibitors is discussed. It is concluded that they exert their effects, not through their weakly oestrogenic activity but through a property common to synthetic progestational compounds.
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Search for other papers by J. A. LORAINE in
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SUMMARY
The effect of the administration of 50 i.u. pregnant mare serum gonadotrophin (PMSG), followed 72 hr. later by 25 i.u. human chorionic gonadotrophin (HCG), on ovarian weight, ascorbic acid and cholesterol has been investigated in rats of the Wistar strain bred in a closed colony.
In the ovarian ascorbic acid depletion (OAAD) test for luteinizing hormone (LH) the bioassay is generally carried out from 5 to 9 days after the administration of HCG. The present investigation has shown that at this time ovarian ascorbic acid levels in untreated animals are in the same range as in rats pretreated with PMSG and HCG.
The reasons for conducting the ovarian cholesterol depletion (OCD) test for LH on the 11th day after treatment with HCG are discussed. It is concluded that in the strain of animal used assays conducted before this time are likely to be unsatisfactory.
It is postulated that the main function of the pretreatment procedure in both assay methods is to produce a large amount of highly reactive ovarian tissue which is readily responsive to the administration of LH.
