The meaning or elevated serum percent free thyroxine (% FT₄) in patients with hyperthyroidism was investigated by the dialysis method. The mean % FT₄ was 0.0340% in patients with hyperthyroidism, and the value was significantly higher than that (0.0178%) in euthyroid controls. The enriching normal sera with cold T₄ to give the same concentration of T₄ as the one in patients with hyperthyroidism resulted in no significant increase of % FT₄ in a phosphate buffer. % FT₄ increased significantly in a barbital buffer not only in normal sera enriched with T₄ but also in hyperthyroid sera. Moreover, % FT₄ in thyroid hormone free sera of hyperthyroid patients prepared by charcoal was 0.0209%, significantly lower than the value in the original one. Serum TBG concentration in patients with hyperthyroidism was 18.4μg/ml, which was not different from that in the euthyroid control, but it increased significantly after treatment with antithyroid drugs. In patients with hyper-thyroidism, the association constant of TBG for T₄ measured by equilibrium dialysis was 3.15 X 10⁹M^-1, and the value was significantly lower than that (7.55 × 10⁹M^-1) in the euthyroid control. The association constant increased significantly after treatment with antithyroid drugs for hyperthyroidism. From these results it is likely that the elevated % FT₄ in sera of patients with hyperthyroidism is ascribed to their decreased association constant of TBG for T₄ as well as to both their diminished TBG concentration and the increased total T₄ concentration.

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Among 281 patients who underwent subtotal thyroidectomy for hyperthyroidism five to ten years ago, recurrent hyperthyroidism was found in 30 (10.7%), T3-toxicosis in 17 (6.0%) and hypothyroidism in 18 (6.4%). The remaining 216 subjects were clinically euthyroid, but a raised level of serum thyroid-stimulating hormone (TSH;>9μU/ml) was found in 100 (35.6%). On 65 of the 216 euthyroid patients, the TRH test, T3 suppression test and measurement of antithyroid antibodies were performed.
The results revealed that 37 of the 65 cases (56.9%) showed an abnormal response to TRH. Nine of these (13.8%) showed either a non-or hyporesponse (peak value of TSH was less than 4.9 μU/ml), and 28 cases (43.1%) revealed a hyper-response (peak value of TSH was more than 35 μU/ml). For the T3 suppression test, 18 of the 65 cases (27.7%) were non-suppressible. All of the 9 non-or hypo-responders showed non-suppressible. For the antithyroid antibodies, 44 of the 65 cases (67.7%) showed microsome test positive and 18 cases (27.7%) revealed positive thyroid test. The positive thyroid test and positive microsome test were more frequent in non-suppressible patients than in suppressible patients. All of the non-suppressible subjects were microsome test positive. These results suggest that there are correlations between antithyroid antibodies, especially antimicrosomal antibodies and T3 non-suppressibility in euthyroid patients with Graves' disease after subtotal thyroidectomy.

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According to recent advances in biochemistry, it is believed that the biologically active site of the parathyroid hormone (PTH) is the amino-terminal portion. Recently human PTH (1-34) (hPTH (1-34)) was synthesized in this country and has been used at a few laboratories. However, the clinical usefulness of this synthetic hPTH (1-34) remains uncertain.
In this paper, whether or not hPTH (1-34) can be used for diagnostic evaluation of hypoparathyroidism and pseudohypoparathyroidism as well as native bovine PTH is investigated.
The subjects were 5 patients with untreated idiopathic hypoparathyroidism (group Al), 10 patients with treated idiopathic or post-operative hypoparathyroidism (group A2), 3 patients with treated pseudohypoparathyroidism (group B) and 6 normal controls (group N).
5, 10, 20 or 30μg of hPTH (1-34) (Niall sequence, Toyo Jozo Inst.) or 200 units of bovine PTH (Parathormone, Eli Lilly) were injected intravenously with saline at 10 : 00 a.m. Urine was collected at 9 : 00, 10 : 00, 10 : 30, 11 : 00, 11 : 30, 12 : 00 and 13 : 00 and blood was driven at 9 : 30, 10 : 15, 10 : 30, 11 : 00, 11 : 30 and 12 : 00. Urine, serum phosphate, calcium and creatinine were measured by Technicon Autoanalizer. Urine and plasma adenosine 3', 5'-monophosphate (cAMP) were measured by radioimmunoassay using YAMASA cAMP kits.
The results were as follows :
1) 5, 10 and 20μg of hPTH (1-34) were given to 2 patients in group Al. The doseresponse relationship was observed as to the maximum increase in the urinary cAMP excretion after hPTH (1-34) administration in both patients. The phosphaturic response to 5μg of hPTH (1-34) was lower than that to 10 or 20μg in one patient. In another, 5, 10 and 20μg of hPTH (1-34) had almost the same phosphaturic effect.
2) The basal excretion of urinary cAMP was significantly lower in groups Al, A2 and B than group N. In groups Al and A2, 20μg of hPTH (1-34) caused an 82.7 or 97.7 fold in-crease in urinary cAMP excretion. In group B, however, it caused only several fold increase.
3) The basal excretion of urinary phosphate was significantly lower in groups A1, A2 and B than in group N. Phosphaturic response to hPTH (1-34) was more than 5 times in groups Al and A2 but very low in groups B and N.
4) The basal excretion of urinary calcium was significantly low in group Al, but almost the same level of excretion was observed in groups A2, B and N. Urinary excretion of calcium after hPTH (1-34) administration decreased in groups A1 A2 and N, while it slightly elevated in group B.
5) The basal plasma cAMP levels were higher in groups A2 and B than in group N, and were not significantly different in groups Al and N. The responsiveness to hPTH (1-34) showed the same tendency to urinary cAMP.
6) Renal responses of cAMP and phosphate to 30pg of hPTH (1-34) were not so different from those to 20pg of hPTH (1-34) in patients with idiopathic hypoparathyroidism and pseudohypoparathyroidism.
7) There was a significant positive correlation between basal urinary phosphate and cAMP excretion.
8) 20μg of hPTH (1-34) and 200 units of Parathormone had almost identical effects on urinary cAMP excretion in patients with idiopathic hypoparathyroidism and pseudohypoparathyroidism, but hPTH (1-34) had less effect on urinary phosphate excretion than Parathormone.
From these results, it is suggested that synthetic human PTH (1-34) can be used for diagnostic evaluation of patients with hypoparathyroidism and pseudohypoparathyroidism as well as native bovine PTH.

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Bovine serum TeBG was purified by 17α-carboxyethynyl-17-hydroxy-4-androsten-3- one sepharose 4B affinity chromatography (T-sepharose) and hydroxylapatite column chromatography. The chemical and immunological properties of the purified TeBG were surveyed and the following results were obtained.
1) The procedures yielded 400μg of protein from 1 L of bovine serum with an overall purification of about 7,560 fold.
2) The purified TeBG was homogenous and was about 80,000 in molecular weight based on sucrose density gradient ultracentrifugation and polyacrylamide gel electrophoresis.
3) The antibody prepared against the purified bovine TeBG did not cross-react with dog and human sera, suggesting the species specificity of TeBG.
4) ¹²⁵I-bovine TeBG, labelled by the lactoperoxidase method, had a high binding capacity to the anti-bovine TeGB antibody.
These results indicate the applicapability of purified TeBG and the antiserum for TeBG RIA.

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We have previously reported that estradiol (E₂) administration induces a particular isozyme of thymidine kinase (TK) in the uterus of immature rats but not of adult rats. This peculiar TK isozyme, which seems to be related to DNA synthesis in myometrial cells induced by E₂ diminishes with the maturation of the rat. Growth with age and hypertrophy during pregnancy are similar phenomena from the standpoint of physiological uterine development. We are interested in the effect of pregnancy on TK activity and the cytoplasmic and nuclear estrogen receptor (ER) in the uterus. In the present study, an oviduct on one side was sectioned previous to mating, in order to compare TK and ER between the non-implanted and implanted uterine horns of the same rat during one to three weeks of pregnancy. No implantation was found in the uterine horn with the sectioned oviduct.
In both the nonimplanted and implanted uterine horns, no increment of TK activity and no appearance of the specific TK isozyme were found during pregnancy. It seems that uterine growth with maturation is not similar to uterine hypertrophy during pregnancy. A local effect of E₂ supplied by trophoblast is not evident in the level of TK.
The cytoplasmic ER level of the nonimplanted horn was approximately twice that of the implanted horn. Unchanged levels of nuclear ER were shown in both uterine horns during pregnancy. The lower concentration of cytoplasmic ER in the implanted horn than in the nonimpalnted horn during pregnancy might be caused by the local effect of E₂ supplied by trophoblast.
These results indicated that uterine development during pregnancy might be caused by an enlargement of individual myometrial cells, and the local effect of steroids supplied by trophoblast might support ovum-implantation to the adjacent endometrium.

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The heterogeneity of human PRL has been demonstrated by gel filtration and subsequent radioimmunoassay of each fraction. Three distinct froms of PRL, peak I (void volume), II and III (monomeric) PRL, were identified after fractionation of serum of a case with PRL secreting pituitary tumor. To further study variations and physiological significance of these different froms of PRL, we observed fractionation patterns of serum PRL before and after TRH administration in normal subjects and in patients with idiopathic galactorrea. In normal subjects, TRH markedly increased monomeric PRL with a resultant relative increase in percentage of peak III or monomeric PRL, whereas peak II PRL increased to a lesser extent and change of peak I PRL was not satistically significant. In idiopathic galactorrhea, the distribution of the three forms of PRL was similar to that of a TRH-stimulated state in normal subjects. TRH administration did not alter the basal pattern and increase in whole serum PRL by TRH stimulation was much less than in normal subjects.
To examine the possibility of conversion of monomeric to larger molecular forms in peripheral circulation, purified ¹²⁵ I-hPRL was intravenously injected to a rat and the plasma samples subsequently obtained were fractionated. In the plasma at 5 min. after injection, 49% of radioactivity was eluted at the void volume on gel filtration. Monomeric ¹²⁵ I-hPRL diminished sharply in plasma samples at following time intervals, and accounted for 3.5% of the recovered radioactivity at 60 min. after injection. Similarly a considerable proportion of 125I-hPRL was converted to the large molecular (peak I) form after incubation in human sera at 37°C for 3 hours. In sera from patients with liver cirrhosis, 8.0 ± 2.2% (mean ± SD) of radioactivity was eluted at the void volume on gel chromatography. In contrast 14.7 ± 1.6% of ¹²⁵ I-hPRL was converted to the peak I form after incubation in sera of normal subjects, exhibiting significantly higher percentages compared with sera from patients with liver cirrhosis.
These results suggest that the distribution of heterogeneous components of human PRL are variable in different circumstances and the biological activity of PRL may not always parallel the whole serum concentration, and that one of the sources of the large molecular form of circulating PRL is peripheral conversion from monomeric PRL.

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Human chorionic gonadotropin (hCG) was extracted and purified from the urine of four patients with hydatidiform mole. The immunological activities of the hCG-hydatidiform mole by hCG radioimmunoassay (RIA) ranged from 9,380 to 9,700 IU/mg, and the biological activities measured by the immature rat ovarian weight method ranged from 7,250 to 7,780 IU/mg.
The results of the amino acid compositions of all the hCG-hydatidiform moles were practically identical with those of hCG-normal pregnancies. The carbohydrate moiety of the hCG-hydatidiform mole was also suspected to be almost similar to that of hCG-normal pregnancies by the results of their in vitro and in vivo biological activities.
It was demonstrated that hCG-hydatidiform mole was composed of α and β subunits (similar to a hCG-normal pregnancy) when hCG-hydatidiform mole was separated into subunits by SDS disc electrophoresis after treatment with mercaptoethanol.
The RIA system of hCG-hydatidiform mole can be established. The concentrations of hCG in sera of normal pregnant women and patients with trophoblastic diseases assayed by hCG-hydatidiform mole RIA were equivalent to those obtained by a standard hCG RIA.
Hence, a standard hCG-immunoassay method used in the management of trophoblastic diseases is considered reasonable so far as the immunoantigenecity of hCG is concerned.

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A solid phase radioimmunoassay (RIA) in which plasma renin activity (PRA) can be measured in a single test tube throughout enzymatic reaction and determination of angio-tensin-I (A-I) generated was developed. Two hundred μ1 of plasma samples was incubated with phenylmethylsulfonyl fluoride (PMSF) and ethylenediamine tetraaceticacid (EDTA) at pH 6.0 or 7.4 at 37°C for 1 hr, then the generated A-I was determined by solid phase RIA under 3 hr of incubation time at room temperature. An antibody adsorbed onto polystyrene beads was used for B/F separation of RIA.
The combination of PMSF and EDTA gave significant inhibition of angiotensinase and converting enzyme. Pepstatin A, proteolytic enzyme inhibitor, was found to be effective to block the renin activity during the incubation for RIA at room temperature. Recoveries of A-I added at two levels of 1.0 and 4.0 ng/ml to plasma sample incubated at 37°C for 1 hr were 102 ± 28.6% and 105 ± 15.2%, respectively. Intra-assay precision (CV) ranged from 2.5 to 10.3% and inter ·assay from 7.0 to 13.7% with four pooled sera. PRA obtained under enzymatic reaction of pH 6.0 showed approximately twice as much as that of pH 7.4. The assay correlated closely with the other established solid phase radioimmunoassay kit (r= 0.996). Normal values of PRA were 1.03 ± 0.78 ng/ml/hr (n=66) at supine position and 1.33 ± 1.15 ng/ml/hr at upright position.
The method for the measurement of PRA, described in this paper, can be clinically used as one of convenient methods from its simplicity and reproducibility.

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