Folia Endocrinologica Japonica
Online ISSN : 2186-506X
Print ISSN : 0029-0661
ISSN-L : 0029-0661
Volume 43, Issue 11
Displaying 1-7 of 7 articles from this issue
  • Akira WATANABE, Katsuharu KIMOTO, Akira WATANABE, Katsuharu KIMOTO, Ha ...
    1968 Volume 43 Issue 11 Pages 1077-1082,1065
    Published: February 20, 1968
    Released on J-STAGE: September 24, 2012
    JOURNAL FREE ACCESS
    In 1958 Nelson and associates described a patient who developed an ACTH-producing tumor of the pituitary gland following bilateral adrenalectomy for Cushing's syndrome with adrenal hyperplasia. Since then similar cases have been reported by other investigators in western countries. It is the purpose of this communication to describe an additional case, which is the first one reported in the Japanese literature : the anterior pituitary of this case, however, showed hyperplasia at necropsy.
    A twenty-nine-year old woman was first admitted to the Second Department of Internal Medicine of Kyushu University Hospital on April 18, 1958, because of obesity of four years' duration.
    Physical examination revealed the classical picture of Cushing's syndrome with plethoric moonface, truncal obesity, purple abdominal striae, hirsutism, ecchymoses and kyphosis. Control values of urinary 17-hydroxycorticosteroids per 24 hours ranged from 12 to 18 mg and after 2 8-hour infusion of ACTH intravenously on 2 successive days, the values rose to 25 and 34 mg. The glucose tolerance curve and serum electrolytes were within normal limits. The blood pressure was 170/112. X-ray examination revealed generalized osteoporosis and bilateral nephrolithiasis. Pituitary fossa was at the upper limits of normal size on X-ray study. Visual acuity and ocular fundi were normal. Perimetry disclosed an irregular defect of lower half-field on the left. Visual field in the right eye was normal.
    Bilateral total adrenalectomy was performed in two stages in August and in October 1958. The left adrenal gland weighed 13 gm, and the right 7 gm. Hyperplasia of the cortex, particularly of the zona fasciculata, was seen in both glands (Fig. 1). The patient was maintained postoperatively on 37.5 mg of cortisone per day orally. Within three months after operation the signs and symptoms of Cushing's syndrome markedly improved. The patient was discharged on hydrocortisone in February 1959 and remained in good health. A few months later she began to work in an office.
    She experienced episodes of adrenal crisis of one to two weeks' duration in April and in December 1960 and in August 1961.
    In spring 1962 she developed gradually a brownish tan over her face, hands and feet. She was admitted to the hospital on August 16, 1962 (3 years and 10 months after the bilateral adrenalectomy), because of cutaneous pigmentation.
    Skull roentgenogram revealed marked ballooning of the sella turcica with slight destruction of the dorsum sellae. Perimetry disclosed bitemporal hemianopia with concentric contraction of the nasal filed in both eyes. The irregular defect of lower half-field on the left seen before the adrenalectomy was still unchanged. Funduscopic examination showed pallor of the disc in both eyes. No abnormalities were seen in the cerebrospinal fluid. The blood pressure was 146/102.
    The patient received radiocobalt therapy for a total estimated tumor dose of 4050 r over the period September 26, 1962, to October 30, 1962. The pigmentation decreased gradually during the month following the cessation of irradiation. Bitemporal hemianopia with concentric contraction of the nasal field in both eyes improved slightly. Ballooning of the sella turcica and pallor of the optic disc in both eyes were unchanged. The patient was discharged from the hospital in December 1962. She worked again in an office, receiving 25 mg of cortisone per day orally.
    In July 1965 she developed abdominal distension, and was admitted to our ward in August 1965. Examinations disclosed gastric cancer with hemorrhagic ascites. She died on November 15, 1965.
    At autopsy the pituitary was enlarged, 1.8 × 1.2 × 0.8 cm in size and 1.7 gm in weight. No nodule was seen. Microscopically the anterior lobe showed hyperplasia which was composed of numerous acidophil cells and scanty chromophobe and basophil cells.
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  • Shugo MURATA
    1968 Volume 43 Issue 11 Pages 1083-1096,1067
    Published: February 20, 1968
    Released on J-STAGE: September 24, 2012
    JOURNAL FREE ACCESS
    Studies were made on the metabolism of two popular progestins, 17α-ethynyl-19-nortestosterone (ENT) and 17α-ethynyl-estrenol (EEL).
    ENT in a dose of 100 mg per day, 3H-ENT or 14C-ENT was administered to post menopausal women who were operated on for uterine cancer. After the administration, urine samples were pooled every 24 hours and stored. The total radioactivity of each 24 hours' urine smple was estimated by counting in a Packard Tricarb liquid scintillation spectrometer (Fig. 1).
    In every three cases, about 30% of radioactivity was found to be excreted in the urine during 5 days. The urines were hydrolysed and extracted as indicated in Table 2. Using Girard-T reagent, the crude extract was further separated into ketonic and non-ketonic fractions. The extract of each fraction was absorbed on alumina and eluted successively with 0.1, 0.3, 0.5, 5.0 and 30% methanol in benzene. From this column, four radioactive peaks were eluted (Fig. 2 and 3).
    The first radioactive peak was eluted in a similar fraction as 17α-ethynyl-5α-19-norandrostane-17β-ol-3-one. However, these two compounds were not identical in Rf values on paper chromatography, silicagel thin layer chromatography (T.L.C.), and from retention time (Rt) in gasliquid chromatography (GL.C.). It was highly probable from this chromatographic behavior that the radioactive metabolite was 17α-ethynyl-5β-19-norandrostane-17β-ol-3-one.
    The second radioactive peak corresponded in the column chromatographic behavior to ENT which was eluted with 0.1 % methanol in benzene. This was applied to a paper and run in benzene-ligroin-methanol-water system. It became a single radioactive peak corresponding in it mobility to authentic ENT. After several repeated paper chromatographies, UV absorption and sulfuric acid chromogen spectra of this radioactive material were measured. Identical spectra of this material with those of authentic ENT were obserbed (Fig. 10). To an aliquot was added carrier ENT and crystallized several times from aqueous ethanol. The specific activity remained constant throughout the above mentioned crystallization (Table 3). These results suggested that this radioactive material was ENT.
    The third radioactive peak was eluted in 0.3% methanol in benzene and was compared to 17α-ethynyl-5α-19-norandrostane-3β, 17β-diol which was obtained from an authentic sample of 17α-ethynyl-5α-19-norandrostane-17β-ol-3-one by its reduction with sodimu borohydride. Paper chromatography, silicagel T.L.C. and G.L.C. were carried out with these two compounds. UV absorption and sulfulic acid chromogen spectra were also measured. All these results indicated the identity of these two compounds (Fig. 12, 13 and 14). Since, in addition to the above results, the oxidation products of the radioactive metabolite were identical with authentic 17α-ethynyl-5α-19-norandrostane-17β-ol-3-one in their chromatographic and spectrometric properties; this radioactive material was identified as 17α-ethynyl-5α-19-norandrostane-3β, 17β-diol.
    The fouth radioactive material was eluted in 5% methanol in benzene from the column and was separated into 4 to 5 radioactive compounds on the paper chromatography in benzene-methanol-water system (Fig. 20).
    A similar experiment with the urine of patients who were administered EEL, revealed that this compound converted in vivo into metabolites identical with those of ENT. This indicated that EEL first metabolized to ENT and further to those metabolites.
    Overall metabolism of EEL and ENT in vivo was indicated as in Fig. 19.
    Relationship between previously reported androgenic activity and the metabolism of EEL and ENT were also discussed.
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  • Shinzo ISOJIMA, Koji KOYAMA, Chiharu TANAKA, Haruo ADACHI
    1968 Volume 43 Issue 11 Pages 1097-1108,1069
    Published: February 20, 1968
    Released on J-STAGE: September 24, 2012
    JOURNAL FREE ACCESS
    In 1965, Wilde et al tried to use radioimmunoassay for estimating human chorionic gonadotropin (HCG) and Bagshawe et al, Odell and Midgley et al. also used this method for assaying HCG and luteinizing hormone (LH). One of our aims in this paper was to determine whether even partially purified HCG (5,000 I.U./mg) could be used for radioimmunoassay or not, and the other was to determine the proper experimental condition for each step of the reactions, in order to get the best results.
    Anti-HCG sera were prepared by giving two injections to four rabbits at 2-week intervals of 1,000 I.U. of crude preparation of HCG (1,000 I.U./mg) in 1 ml of phosphate saline buffer, freshly emulsified with 1 ml of complete Freiund's adjuvant (Difco) each time. The injections were made in the footpads and i.d. After 3 weeks of rest, a series of booster injections with 1,000 I.U. of HCG without adjuvant were given i.p. 5 times every other day. Rabbist were bled 7 days after the last injection and the sera were obtained. The antibody titers were from 1 : 6,400 to 1 : 25,600 by hemagglutination.
    For preparing absorbed anti-HCG serum, one ml of antiserum was incubated with 30 mg of an alcohol precipitate of urine from a child and 30 mg of human serum protein at 37°C for 60 min., and was kept at 4°C over-night. After centrifuging the mixture, the supernatant, the absorbed anti-HCG serum, showed only one precipitin band to HCG antigen on immuno-electrophoresis. Radioiodination of HCG by 131I was carried out by the modified chloramine T method of Greenwood.
    All materials were diluted with 0.5% bovine serum albumin (BSA) in borate buffered saline (BBS) -diluent-, and the 2nd IRP-HMG and Pergonal 500TM were used as standard LH.
    In radioiodination, the ratio of chloramine T to HCG was 0.4-1.3 : 1. The high specific activity of 131I-HCG which was more than 200 mc/mg, was obtained, and around 60% of the 131I-HCG preparation was precipitable with absorbed anti-HCG serum.
    The procedure of radioimmunoassay was as follows : The diluted anti-HCG serum was mixed with sample (HCG) and 131I-HCG. After incubation, the inactivated diluted rabbit serum (as carrier protein) and horse anti (rabbit serum r-globulin) serum globulin were added and then incubated. Th resulting precipitate was washed once with diluent and hydrolyzed with 1 ml of 2N NaOH and then counted with Scintillation Counter.
    Th proper experimental conditions for each step of radioimmunoassay were determined. 1. The combination of 131I-HCG and anti-HCG needed 48 hrs. for the maximum reaction (1 st reaction). 2. Cold HCG (sample) should be added to anti-HCG serum at least 4 hrs. before the addition of 131I-HCG in the 1st reaction. 3. The proper temperature during incubation in the 1st and 2nd reaction was 4°C. 4. As diluent, 0.5% bovine serum albumin in BBS was necessary and enough. 5. In the 2nd reaction, 0.1 ml of diluted (1 : 2,000) rabbit serum as carrier protein, gave the maximum precipitate with 0.2 mg of our horse anti (rabbit serum r-globulin) serum globulin. 6. The 2nd reaction needed more than 6 hrs. at 4°C.
    The standard curve of HCG shows that the amount of HCG from 0.0025 I.U./ml to 0.3125 I.U./ml was sufficient to be estimated quantitatively and the amount as much as 0.001 I.U./ml of HCG was also qualitatively detectable by this method. The amount of HCG in the urine was measured simultaneously by radioimmunoassay, hemagglutination inhibition rection and bioassay. The results showed that the radioimniunoassay could be clinically used. This method was also very useful to detect the low titer of HCG in the urine which could not be detected by other methods.
    The amount of LH in the urine of adult women having normal ovarian cycle, was radioimmunologically assayed, and it was shown that the peak of LH appeared one to three days before ovulation assumed by B.B.T.
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  • Masaji INOUE, Yoichi SUGIHARA, Motozo KO, Toru OISHI, Shigeru TOMITA
    1968 Volume 43 Issue 11 Pages 1109-1112,1071
    Published: February 20, 1968
    Released on J-STAGE: September 24, 2012
    JOURNAL FREE ACCESS
    1) Estradiol showed the pregnancy period prolonging effect and the inhibitory effects on mechanic and electric activities of myometrium in vitro. Those inhibitory effects of estradiol could not be explained by the change of resting potentials of myometrial cells, but by the suppression of the ACh binding with depot protein and receptor.
    2) Estradiol inhibited the accelerating effect of oxytocin on the binding of ACh with depot protein and receptor. These fact coincide with the inhibitory effect of estradiol on myometrial contractions induced by oxytocin.
    3) Considered from the standpoint of ACh, the accelerating effect of estradiol on myometrial contractions may depend on the increase of myometrial ACh after administration of estradiol.
    4) In conclusion, it is emphasized that estradiol has two functional faces to accelerate and inhibit myometrial contractions.
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  • Masaji INOUE, Motozo KO
    1968 Volume 43 Issue 11 Pages 1113-1116,1073
    Published: February 20, 1968
    Released on J-STAGE: September 24, 2012
    JOURNAL FREE ACCESS
    1) Progesterone showed the early effect that myometrial contractions were temporarily promoted by progesterone in vitro. The effect is very interesting, because progesterone accelerated the binding of ACh with depot protein and receptor.
    Progesterone in vitro had no effects in the early stage on oxytocic actions of accelerating myometrial contractions and the binding of ACh with depot protein and receptor.
    2) As the time elapsed progesterone continuously accelerated the binding of ACh with depot protein; however it disturbed ACh on its binding with receptor. These results may relate to the inhibited myometrial contractions which followed the early accelerated contractions under the influence of progesterone in vitro.
    Progesterone weakened oxytocic actions on myometrial contractions and the binding of ACh with tissue protein, as the late effects.
    3) The labor provoking mechanism was explained from the standpoint of myometrial ACh.
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  • Takao UEMURA
    1968 Volume 43 Issue 11 Pages 1117-1137,1075
    Published: February 20, 1968
    Released on J-STAGE: September 24, 2012
    JOURNAL FREE ACCESS
    The direct actions of some sex steroids on ovarian functions were studied using the longterm administration of those steroids to female albino rats, and the ovarian cholesterol and ascorbic acid contents before and after the combined administration of some gonadotropins (PMS and HCG) were determined.
    Form the experimental data of mature female rat with regular sexual cycles, I found that the long-term sadminitration of estrogen or progesterone inhibited the sexual cycle not only through the hypothalamico-pituitary system but also through the direct action of these steroids to the ovary itself.
    I obtained the same data with the experiment using immature female albino rat without sexual cycles.
    From these experimental data it was concluded that the ovarian sex steroids have some direct regulatory or inhibitory action on the ovary itself, especially on its steroidogenesis.
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  • Takao UEMURA
    1968 Volume 43 Issue 11 Pages 1138-1153,1076
    Published: February 20, 1968
    Released on J-STAGE: September 24, 2012
    JOURNAL FREE ACCESS
    In the first report, it was demonstrated that the long-term administration of sex steroids (estrogen and progesterone) inhibited the ovarian function not only by the so-called feed back mechanism but also by the direct action on ovaries of mature and immature albino rats.
    In order to determine the direct action of those steroids on ovaries exactly, in this experiment, I used the hypophysectomized female albino rat without the endogenous gonadoropin release. After the long-term administration of estradiol benzoate or progesterone, combined gonadotropin injection could not induce the ovarian cholesterol and ascorbic acid depletion, which were detectable in the control animal group without the sex steroid administrations.
    From these experimental results, I conclude that under some conditions ovarian sex steroids, namely estrogen and progesterone, control the steroidogenesis in ovaries themselves through the reduction of reacitivities to gonadotropic actions.
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