One hundred microgram of luteinizing hormone releasing hormone was intravenously injected into 19 female patients with systemic lupus erythematosus and plasma luteinizing hormone levels before and after the injection were determined with a method of solid phase radioimmunoassay.
In 4 patients among them, the basal levels of luteinizing hormone were abnormaly high. A question about the existence of autoantibodies to luteinizing hormone in the patient's plasma raised to explain the reason for the high level of luteinizing hormone. The results from 3 experiments for solution of this question were as follows.
1) The patient's plasma was proved to have a binding activity with labeled luteinizing hormone.
2) This binding activity is specifically inhibited by purified luteinizing hormone.
3) This binding activity was located in γ-globulin fraction. From the results, the existence of autoantibodies to luteinizing hormone in the 2 patients' plasma was concluded.

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We have already reported a high rate of occurrence of antimicrosomal antibodies in diabetes mellitus.
Thirteen of 507 diabetics (2. 5%) were positive with antithyroglobulin antibodies and thirty-one (6. 1%) were positive with antimicrosomal antibodies compared to 2. 3% and 2.5% respectively in normal controls.
Two of 34 insulin dependent diabetics (5. 9%) were positive with antithyroglobulin antibodies and ten (29.4%) were positive with antimicrosomal antibodies compared to 2. 3% and 4.4% respectively in 473 insulin independent diabetics.
To clarify the association of insulin antibodies and thyroid antibodies in diabetics, antithyroid antibodies in 507 diabetics were tested by tanned red cell hemagglutination test and insulin antibodies were demonstrated by using a method descrived by Wright in a modified form.
Twelve of 482 diabetics negative-insulin antibody (2. 5%) were positive with anti-thyroglobulin antibodies and thirty (6. 2%) were positive with antimicrosomal antibodies.
Only one of 25 positive-insulin antibody (4 %) was positive with antithyroid antibodies respectively.
No evident correlation was observed between antithyroid antibodies and insulin antibodies.

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The purpose of this study is to clarify the significance of synthetic LH-RH (RH) as a therapeutic agent in the treatment of anovular women who fail to respond to clomiphene therapy.
Methods : Fifteen women with hypofunction of the hypothalamus-pituitary system who were given the RH-test were divided into two groups according to the maximum increased level of LH shown by each. Group I consisting of 7 women who showed increased levels of over 20 miu/ml received a combination of clomid + estrogen + RH (C.E.R.). Blood levels of LH, FSH and estradiol were measured in each. Group II consisting of 8 women who showed maximum levels of LH below 20 miu/ml received RH intramuscularly, 100 μg per day for 20 to 30 days (referred to as RH treatment). The RH-test was performed in this group before treatment, and 1, 7 and 21 days after treatment. Blood levels of estradiol were also determined before, during and after treatment. Results : 1) Ovulation occurred in 5 of the 7 women in group 1 and 3 became pregnant. The LH surge was prolonged and hormonal secretion increased with administration of C.E.R.. The normal estrogen surge and gonadotropin surge occurring in the ovulatory cycle were thus produced artificially. Since administration of clomiphene alone tends to produce insufficiency of cervical mucus secretion because of its anti-estrogen effect, the addition of estrogen to the combination of C.E.R. probably increases the cervical secretion of mucus and enhances the probability of impregnation.
2) Compared to pretreatment levels, RH treatment clearly increased the pituitary secretion of LH and FSH. Even 3 weeks after RH treatment, the pituitary secretion of gonadotropin was definitely higher than before treatment. Fluctuations in blood levels of estradiol during and after treatment were not significantly different than before treatment. These observations would suggest that prolonged administration of RH enhances the gonadotropic function of the pituitary, not only in the secretion of the hormones but also in the synthesis of gonadotropin. Judging from the insignificant effect on the blood level of estradiol, it would appear that RH itself possesses the ability to produce gonadotropin.

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In order to assess the pathophysiology of gonadotropin secretion in patients with idiopathic precocious puberty, responses of LH and FSH to LH-releasing hormone (LH-RH), clomiphene citrate (Clomid) and medroxyprogesterone acetate (Depo-provera) in 6 female patients were tested.
The basal levels of serum LH and FSH were elevated in 4 of 6 patients and enhanced LH response to 100 μg of intravenous LH-RH was observed in 5 patients, while FSH response was comparable to that of prepubertal subjects. Administration of Clomid (50 mg) for 7 days provoked LH increase in 3 out of 6 patients in contrast to no LH change found in normal prepubertal girls, while FSH levels decreased in 3 out of 6 patients. Intramuscular injection of Depo-Provera (100 mg) reduced LH and FSH levels in 4 patients. The Depo-Provera therapy in a dose of 75-225 mg every two weeks, caused suppression of breast development and menstrual bleeding but the prevention of growth spurt and bone age maturation were insufficient. Basal boby temperature charts in 2 patients showed an ovulatory pattern immediately following cessation of Provera therapy of more than 8 years' duration.
In conclusion, it is suggested that 1) function of hypothalamo-pituitary-ovarian system in patients with idiopathic precocious puberty is similar to that of normal pubertal subjects, 2) therapeutic doses of Depo-Provera, enough to suppress the patients' secondary sex characterics, was not sufficient to control bone age maturation 3) it is neccessary to follow up patients more closely by measuring serum levels of gonadotropin and estrogen during the course of therapy.

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Composite microassays for plasma progesterone (P), 17α-OH-progesterone (17P), estrone (E₁), 17β-estradiol (E₂), and estriol (E₃) were developed. Steroids were extracted from plasma samples with diethyl ether, and were separated from each other through two steps of sephadex LH-20 microcolumn chromatography (benzene : methanol 85 : 15, and n-hexane : benzene : methanol 80 : 10 : 10), prior to assays by radioimmuoassay (P, E₁, E₂, and E₃) and competitive protein binding assay (17P). Steroids were recovered satisfactorily through these procedures (mean recovery ; P : 107. 6%, 17P : 102. 3%, E₁ : 88.2%, E₂ : 84. 1% E₃ : 78. 5%). The detectable range for P, 17P, E₁, E₂, and E₃ were 0.01-2 ng/tube, 0.1-10 ng/tube, 10-2000pg/tube, 10-2000 pg/tube, and 0.5-100 ng/tube. The interassay coefficient of variations were less than 10. 7%, 15. 2%, 17. 8%, 11. 4%, and 28. 1%, respectively.
These steroids were measured daily in 4 menstrual cycles from 4 normally menstrauting women. Plasma FSH and LH were quantitated previously¹²). The following results were obtained ; 1) Plasma P elevated from around LH surge (Day 0), and reached a peak on day +5 - +7 with the values of 12. 74 ± 2. 34 ng/ml (Mean ± S.D.). 2) Slight decrease in P levels was noted on day +8 - +9. 3) A peak in 17P was observed in the preovulatory phase (day -3 - -1) with the values of 0.6 - 1. 3 ng/ml in three of four cases. 4) Changes of 17P during the luteal phase were paralleled to those of P with a peak of 1. 16 ± 0. 31 ng/ml on day +5 - + 7. 5) No remarkable patterns were found in E₁ levels throughout the menstrual cycle. 6) A sharp peak in E₂ was detected in the preovulatory phase (day - 1-0) with the values of 709. 2 ± 95. 9 pg/ml. 7) The second peak of E₂ with 378. 6 ± 140. 7 pg/ml was observed in the late luteal phase (day +8 - +12). 8) E₃ was not detected in all samples.
The interrelationship between steroids and the correlation with the morphological changes of the ovaries in the normal menstrual cycle are discussed. In the follicular phase, the theca interna cells around the maturing follicle may be growing under the influence of pituitary gonadotropins to secrete large amounts of 17P and E₂, which may possiblly affect the pituitary for LH surge, followed by ovulation. In the luteal phase, both the granulosa cells and theca interna cells are luteinized, which may produce and secrete large amounts of P, 17P, and E₂.

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Plasma progesterone, 17α-OH-progesterone, estrone, 17β-estradiol and estriol in human normal pregnancy, labor and the puerperium were measured simultaneously with radioimmunoassay or competitive protein binding assay. The steroids were extracted from the sample plasma with diethyl ether, and were separated through two steps of sephadex LH-20 microcolumn chromatography prior to the assays.
From the beginning to the 25th week of pregnancy, a gradual rise in the levels of plasma progesterone was noted, followed by a steep increase toward term with a level of 150-250 ng/ml at the end of gestation.
The levels of 17α-OH-progesterone during the 7 th or 8 th week of pregnancy were 2-3 ng/ml, which was above the normal range in the luteal phase, followed by a gradual decrease during 15-25 weeks of gestation. Thereafter, an increase toward term was seen with a mean level of 6-10 ng/ml in the 40th week.
From the beginning to the end of pregnancy, plasma levels of estrone, 17β-estradiol, and estriol increased from less than 0.5 ng/ml to 2-4 ng/ml, from 1-2.5 ng/ml to 6-10 ng/ml, and from less than 1 ng/ml to 6-10 ng/ml, respectively.
The disappearance of steroids from maternal peripheral blood in the puerperium was rapid. The half time of progesterone, 17α-OH-progesterone, estrone, 17β-estradiol, and estriol were 72 min, 68 min, 27 min, 27 min, and 48 min, respectively.
Analysis of the interrelation between several kinds of steroids in pregnancy, labor, and the puerperium with simultaneous microassays will contribute to the clarification of some important physiological and pathological aspects of feto-placental-maternal functions and steroidogenic functions of the corpus luteum in pregnancy

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Ultrastructural changes in the rat endometrium under several conditions including experiments of the inhibition of implantation were studied to clarify the sequential changes of the endometrium and their endocrinologic background during ovoimplantation. The following pertinent findings were noted. 1) Normal pregnancy.
In the luminal surfaces, the microvilli were short and arranged rather irregularly and lost their so-called glycocalyx on L3. The large fungus-like protrusions appeared on L4. On L5, the luminal surfaces were completely covered with small serrated cytoplasmic protrusions. They formed the interdigitation between the apposed surfaces and the uterine lumen was disappeared when the uterus was fixed in situ by the arterial perfusion method. High ATP-ase activity was demonstrated at the site of interdigitation. In the apical cytoplasm, many apical vesicles, well developed Golgi apparatuses, and dilated cisternae of the granular endoplasmic reticulum were observed on L3. On L5, however, these apical vesicles decreased in number and the Golgi apparatuses were small. On the contrary, the semilunar and coated vesicles were numerous, and the lysosomes increased both in number and size. Acid phosphatase activity was detected in these lysosomes.
In the glandular epithelium, progressive increase in the number of small granules and the amount of amorphous substance were observed in the glandular lumen from L2 to L4. But on L4, some autophagosomes had already appeared in the cytoplasm. Swollen and degenerated mitochondria as well as many vacuoles were recognized on L5.
In the stromal cells, the cytoplasm was enlarged, and contained many polyribosomes, granular endoplasmic reticulum with markedly dilated cisternae, lysosomes and lipid droplets on L5. The cytoplasmic protrusions on L5 were elongated and showed various features ; one of them was situated very closely to the basement lamina of the luminal epithelium, and the other surrounded the lumen. Some of these protrusions were connected with the desmosomes. High ATP-ase activity was localized in these cytoplasmic protrusions, on the surfaces of the stromal cells, and in the stroma itself.
2) Delayed implantation and estrogenic transformation.
In delayed implantation, the luminal surfaces were covered with regular microvilli and several fungus-like cytoplasmic protrusions. The cytoplasm contained many apical vesicles and large Golgi apparatuses that were observed both on L3 and L4 in normal pregnancy. At 48 hrs after the injection of estrogen, the interdigitation between the apposed luminal surfaces was also established.
In the glandular epithelium, a slight enlargement of the glandular lumen and the appearance of small granules similar to those observed in normal pregnancy were noted at the 4th hr of estrogen treatment.
In the stromal cells, two cells apposed each other very closely at the 4th hr and formed large cell masses at the 48 th hr of estrogen treatment. The enlargement of the cytoplasm with concomitant development of the organelles was elicited at the 48th hr after the injection of estrogen only when progesterone was administered at the 24th hr after the previous injection of progesterone.
3) No progesterone administration after the ovariectomy on L2.
The microvilli in the luminal surfaces were short and their arrangement was sparse. No fungus-like protrusion was noted. In the cytoplasm, only a few apical vesicles and small Golgi apparatuses were observed. Some secretory granule-like structures were noted in the glandular epithelium. After the injection of progesterone, similar features to those in delayed implantation were recognized.
4) Experiments for the inhibition of implantation.
A) Pretreatment with actinomycin D.
When actinomycin D was administered before the injection of estrogen, nucleolar segregation was recognized in the luminal epithelium, glandular epithelium and the stromal cells.

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