Folia Endocrinologica Japonica
Online ISSN : 2186-506X
Print ISSN : 0029-0661
ISSN-L : 0029-0661
Volume 49, Issue 1
Displaying 1-6 of 6 articles from this issue
  • Shoshi SHIMOYAMA, Toshihiko MIHARA, Sekiji MORITA, Isao MATSUOKA, Kiic ...
    1973 Volume 49 Issue 1 Pages 11-16,1
    Published: January 20, 1973
    Released on J-STAGE: September 24, 2012
    JOURNAL FREE ACCESS
    It has been confirmed by many reports that glucose-6-phosphate dehydrogenase (G-6-PD) activity of the erythrocytes increased in hyperthyroidism. However, the comparison of this activity with the results of thyroid function tests used commonly at present, that is, serum protein-bound iodine (PBI), resin sponge uptake of 131I-T3 (Triosorb test), basal metabolic rate (BMR) and thyroid 131I uptake have not been explored adequately.
    To clarify this problem, erythrocyte G-6-PD activity and the various thyroid functions were measured simultaneously in patients with hyperthyroidism, hypothyroidism and struma nodosa.
    The series comprised inpatients with hyperthyroidism (15), with hypothyroidism (4), with struma nodosa (2) and 10 healthy control subjects. G-6-PD activity of the erythrocytes was determined by the method of Konberg and Horecker in which the amount of NADPH2 newly produced in vitro was determined. The procedures of the determination were performed at 25°C. The effect of hemoglobin concentration in samples was excluded. The enzyme activity was expressed as the change rate ΔOD/min./gHb.
    1. The mean value for G-6-PD activity of the erythrocytes obtained in patients with hyperthyroidism was 10.8±1.38 ΔOD/min./gHb (M±SD), whereas in healthy control subjects, it was 7.26±0.58 ΔOD/min./gHb (M±SD). The difference between these values was statistically significant (p<0.001).
    2. A positive correlation between G-6-PD activity of the erythrocytes and Triosorb test, PBI, BMR and thyroid 131I uptake (24 hours value), respectively, was observed in the patients with hyperthyroidism, hypothyroidism and struma nodosa, an individual correlation coefficient “r” between G-6-PD activity and each one of the other tests was 0.844 (p<0.01, N=21), 0.726 (p<0.01, N=19), 0.880 (p<0.01, N=16) and 0.673 (p<0.01, N=18), respectively.
    3. From the above results, it is suggested that the determination of G-6-PD activity of the erythrocytes can be used as one of the useful thyroid function tests.
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  • Morihiko KAKITA
    1973 Volume 49 Issue 1 Pages 17-31,2
    Published: January 20, 1973
    Released on J-STAGE: September 24, 2012
    JOURNAL FREE ACCESS
    Attempts have been made to study the effects of a hormone controlling the proliferation and differentiation of the endometrial tissue on DNA synthesis by the target cells as well as the working mechanism, the results are as follows :
    1. Estrogen initiates the synthesis of DNA by the epithelial cells of the endometrium as early as at the outset of its action, bringing about a rise in the activity of thymidine kinase in approximately 18 hours and the eventual synthesis of DNA after 24 hours. The reaction of the interstitial cells, however, lacks uniformity as compared with the epithelial cells.
    2. When the synthesis of protein in the uterine tissue was inhibited, most of the epithelial cells shifted to the synthetic stage of DNA. This result has led us to believe that the function of certain specific proteins are in the shift from GI-stage to S-stage. It is believed that the half-life of this protein is about 16 hours.
    3. Progesterone has been proved to possess a potent effect inhibiting the synthesis of DNA. This inhibitory effect is produced through the conjugate action with said specific protein, and adrenaline has been proved to be involved in the working mechanism.
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  • Terukazu TAKANO, Fujio NUMANO, [in Japanese]
    1973 Volume 49 Issue 1 Pages 32-47,3
    Published: January 20, 1973
    Released on J-STAGE: September 24, 2012
    JOURNAL FREE ACCESS
    Analysis of estrogens in urine provids much important information on the endocrinological conditions of man related to his morbid conditions, the more precise method of which, however, is required today.
    In the measurement of estrone, estradiol-17β and estriol several extracting methods, containing gas chromatographic analysis were reported and have been commonly used, However it was recently found by gas chromatography-mass spectrometry (Shimadzu LKB-9000) by the authors that they were not all adequate, producing too much contamination, so that we introduced a new methods in which each urinary estrogen can be accurately analysed with gas chromatography (Shimadzu GC-5APFE). With this method, estrone, estradiol-17β 78 and estriol were certified to be analysed in almost 100% in purities. In this paper our new extraction technique of estrogens from the human urine is reported.
    Urinary estrogens were extracted as follows ; 300 ml of urine collected during 24 hrs. was used. After mixing with 210 g of ammonium sulfate and stirring for 40 min, urine was centrifuged at 8,000 × g for 40 min at 4°C. Precipitated urine was dissolved with 50 ml of water, adjusted with 0.2 M acetate buffer to pH 4.5, added to 100,000 Fishman Unit/800,000 Roy Unit of β-glucuronidase/aryl-sulfatase (Boehringer) and then hydrolysed for 21 hrs. at 37°C. Then, after adding 6 g of NaCl, estrogens were extracted twice with 100 ml of ether which were afterwards washed with 40 ml of sodium carbonate buffer solution (pH 10.5) and with 10 ml of saturated NaHCO3 solution. After the evaporation, the extract was solved with 2 ml of methanol, and fractionated five times by an anion exchange column chromatography using AG1-X2 resin made by Bio-Rad. That is, after being washed with 20 ml of 0.5 M NaHCO3 solution and 25.ml of methanol, the column was poured by 2 ml of estrogen containing methanol for five times. Then the column was washed with 15 ml and 25 ml of methanol to eliminate neutral steroids or impurities.
    Estrogens were then extracted by 30 ml of 80% methanol and 20 μg of cholesterol was added as an internal standard, evaporated to dryness under 40°C. Trimethylsilylation was performed with N, O-Bis- (Trimethylsilyl) -acetamide to analyse estrogens with gas chromatography equipped with 1.5% OV-17 column. Gas chromatography-mass spectrometric analysis was also performed in parallel to identify estrogens. During these procedures several optical conditions were examined.
    1) The optimum hour for hydrolysation of estrogens with β-glucuronidase/aryl-sulfatase was studied in 18, 20, 22, and 24 hours and it was found that hydrolysation for 21 hours exhibited the most effective recovery. The recoveries obtained in this condition are as follows ; estrone-3-sulfate 70.7±2.4%, estrone-3-glucuronide 74.5±3.8%, estradio1-17β3-glucuronide 74.8±3.5%, estriol-3-glucuronide 71.3±4.9% and estriol-16-glucuronide
    2) The optimum unit of β-glucuronidase/aryl-sulfatase in hydrolysed for 21 hrs. was compared among 50,000/400,000,100,000/800,000,200,000/1,600,000,400,000 Fishman Unit/3,200,000 Roy Unit. More than 100,000 F.U./800,000 R.U. of enzyme hydrolysed more than 80.2±5.5% of conjugated estrogens.
    3) The studies on recoveries of the concentration of various estrogen conjugates (0.5, 1.0, 2.5 and 5.0 μg) in the above-mentioned optimum conditions showed as follows;
    estrone-3-sulfate 72.7±4.6%
    estrone-3-glucuronide 83.1±6.3%
    estradio1-17β-3-glucuronide 71.9±2.0%
    estriol-3-glucuronide 80.0±4.4% and estriol-16-glucuronide 79.9±2.9%
    The total recoveries of all of the estrogen conjugates exhibited 78.2±3.6% with this method.
    Gaschromatography-mass spectrometric analysis revealed that the following estrogens were certified to be extracted with this method by gas chromatography;
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  • Osamu YASHIMA
    1973 Volume 49 Issue 1 Pages 48-59,5
    Published: January 20, 1973
    Released on J-STAGE: September 24, 2012
    JOURNAL FREE ACCESS
    Dopamine-β-Hydroxylase is an enzyme which catalyzes dopamine to noradrenaline in final step of catecohlamine synthesis.
    This study was undertaken to determine the human serum Dopaminel-β-Hydroxylase activity and examine effects of fusaric acid (5-butylpicolinic acid) on this enzyme activity in vitro and in vivo.
    A sensitive method for measurement of the human serum Dopamine-β-Hydroxylase activity was established and it was important for the assay to adjust PH of reaction mixtures to PH 4.5.
    The activity ranged from 39.95 to 176.00, mean was 82.31±33.11 (M. ±S.E.) in normal and that of patients with essential hypertension ranged from 31.11 to 234.97, mean was 109.87±57.23 (M.±S.E.) n moles/0.5 ml serum/15 min.
    Fusaric acid showed 50% inhibition of the human serum Dopamine-β-Hydroxylase activity at concentration of 10-7 M in vitro and the activity in patients with essential hypertension was significantly decreased in a few days after the administration of fusaric acid 150 mg per day.
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  • Hideyuki BABA
    1973 Volume 49 Issue 1 Pages 60-79,6
    Published: January 20, 1973
    Released on J-STAGE: September 24, 2012
    JOURNAL FREE ACCESS
    Plasma testosterone levels were measured by radioimmunoassay in 43 normal subjects and in 108 patients with various endocrine disorders. They consist of patients with panhypopituitarism, Kallmann's syndrome, Fröhlich's syndrome, idiopathic hypogonadotropic hypogonadism, Sheehan's syndrome, pituitary dwarfism, Klinefelter's syndrome, ovarian dysgenesis, Cushing's syndrome, Addison's disease, primary aldosteronism, congenital adrenal hyperplasia, 17α-hydroxylase deficiency, hyperthyroidism, primary hypothyroidism, idiopathic diabetes insipidus, acromegaly, precocious puberty, anorexia nervosa, simple obesity and idiopathic hirsutism. Base line plasma testosterone levels ranged from 273 to 1131 ng/100 ml with a mean of 568±208 (SD) in 25 normal males aged from 14 to 67, and from 20 to 56 ng/100 ml with a mean of 33±14 (SD) in 18 normal females aged from 17 to 53.
    In patients with secondary hypogonadism due to panhypopituitarism, Kallmann's syndrome, Fröhlich's syndrome, idiopathic hypogonadotropic hypogonadism, Sheehan's syndrome and pituitary dwarfism, plasma testosterone levels were lower than normal or low normal in both males and females. In patients with primary hypogonadism, plasma testosterone levels were lower than normal or low normal in Klinefelter's syndrome, and normal or low normal in ovarian dysgenesis.
    In female patients with Cushing's syndrome due to adrenocortical hyperplasia, plasma testosterone levels were higher than normal. They were suppressed by 8 mg of dexamethasone and increased slightly by 1 mg of ACTH-Z. On the contrary, plasma testosterone levels in female Cushing's syndrome due to adrenal tumor were normal or lower than normal. They results suggest that the estimation of peripheral plasma testosterone could be helpful in determining the etiology of Cushing's syndrome in female patients.
    In patients with Addison's disease, plasma testosterone levels were normal in males and were significantly low in females.
    The mean plasma testosterone level in female patients with anorexia nervosa was higher than the normal mean in contrast with significant low levels of plasma testosterone in patients with Sheehan's syndrome.
    In female patients with simple obesity, plasma testosterone levels were higher than normal and were suppressed to the normal range by l mg of dexamethasone.
    Plasma testosterone levels were increased significantly after administration of HCG, 4,000 units daily for 3 days intramuscularly, in normal males. They were not increased in male patients with hypogonadism of any etiology. It was possible to differentiate more clearly the patients with hypogonadism with border-line testosterone levels from normal subjects by a response of plasma testosterone to HCG administration. However, it was impossible to differentiate the secondary hypogonadism from the primary hypogonadism by present way of administration of HCG.
    Clomiphene citrate, 150 mg daily for 3 days administered orally, stimulated the pituitary-Leydig cell axis in normal male subjects. On the 2nd day after cessation of administration of clomiphene, plasma testosterone levels were increased by 50% while plasma LH levels were increased by 110%. In a male patient with Frohlich's syndrome clomiphene stimulated neither plasma LH nor testosterone secretion.
    Intravenous injection of 100 μg of synthetic LH-RH induced a significant rise in plasma LH by 350% and plasma testosterone by 120% in normal males. The peaks of plasma LH were observed between 20 and 45 min. after i.v. injection of LH-RH while the peaks of testosterone were observed between 20 to 60 min. In a male patient with Frohlich's syndrome, plasma testosterone level was not increased after injection of LH-RH, although plasma LH was increased significantly. In patients with Klinefelter's syndrome,
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  • Jun HASHIDA
    1973 Volume 49 Issue 1 Pages 80-97,8
    Published: January 20, 1973
    Released on J-STAGE: September 24, 2012
    JOURNAL FREE ACCESS
    In order to analyse the pathogenesis of hypertension, spontaneously hypertensive rats (SHR) were compared with normotensive Wistar strain rats at the age of 8-10 weeks and over the 6 months. Endogenous catecholamine in adrenal glands and brain were determined in the young and adult animals. The levels of tritium labeled 3, 4-hydroxyphenylalanine (3H-DOPA), 3H-dopamine and 3H-noradrenaline in brain stem were determined at various time such as 1, 6 and 12 hours after administration of 3H-DOPA in adult SHR and control Wistar strain rats.
    The other experiments were made by comparing the 3H-DOPA metabolism in SHR treated with reserpine, α-methyl DOPA or 2- (2, 6-dichlorophenylamino) -2-imidazoline hydrochloride [ST-155] with control SHR over the 4 months of age.
    1) The blood pressure and catecholamine levels in the adrenal glands were higher in SHR than control Wistar strain rats at 8-10 weeks or over 6 months of age.
    2) The noradrenaline content was decreased in brain stem of SHR, but was increased in hemispherium and celleberum of them compared with control Wistar strain rats over 6 months of age.
    3) Biosynthesis of 3H-dopamine, 3H-noradrenaline in brain stem of SHR after administration of 3H-DOPA was compared with those of Wistar strain rats. The levels of 3H-DOPA, 3H-dopamine and 3H-noradrenaline were lower in SHR than Wistar strain rats at 1 or 6 hours after administration of 3H-DOPA. The observed decrease of DOPA uptake and catecholamine synthesis in brain stem were considered as suggesting upon the relation to the pathogenesis of hypertension.
    4) Reserpinized SHR showed higher level of 3H-DOPA in brain and heart, but showed lower level of 3H-dopamine, 3H-noradrenaline and 3H-adrenaline in brain, heart and particularly in adrenal glands.
    It is considered that the uptake of 3H-dopamine and synthesis of 3H-catecholamine were inhibited in each organs, and the process from DOPA to dopamine was inhibited in brain and heart of SHR treated with reserpine.
    5)3H-DOPA of each organs (brain, heart and adrenal glands) showed higher levels, but 3H-dopamine and 3H-noradrenaline showed lower levels in SHR treated with α-methyl DOPA. It is considered that α-methyl DOPA inhibits the metabolic process of DOPA to dopamine, and replaces and releases catecholamine in each tissue.
    6) 3H-DOPA, 3H-dopamine, 3H-noradrenaline and 3H-adrenaline (especially in adrenal glands) in brain, heart and adrenal glands showed no change in SHR treated with ST-155.
    It is considered that ST-155 does not influence catecholamine metabolism.
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